ovalbumin and Hypersensitivity

Allergic diseases, in particular atopic asthma, have been on the rise in most industrialized countries for several decades now. Allergic asthma is characterized by airway narrowing, bronchial hyperresponsiveness, excessive airway mucus production, eosinophil influx in the lungs and an imbalance of the Th1/Th2 responses, including elevated IgE levels. Most available interventions provide only short-term relief from disease symptoms and do not alter the underlying immune imbalance. A number of studies, mostly in mouse models, have shown that Mycobacterium bovis bacillus Calmette-Guérin (BCG) treatment is capable of preventing or reducing an established allergen-driven inflammatory response, by redirecting pathogenic Th2 towards protective Th1 and/or regulatory T cell responses. Dendritic cells stimulated by BCG appear to be a crucial first step in the immunomodulatory effects of BCG. While the protective and therapeutic effects of BCG against allergy and asthma are well documented in animal models, they are less clear in humans, both in observational studies and in randomized controlled trials. The purpose of this article is to provide an up-to-date overview of the available evidence on the anti-allergy, in particular anti-asthma effects of BCG in mice, rats and humans.

Topics: Animals; Asthma; BCG Vaccine; Hypersensitivity; Lung; Mice; Mycobacterium bovis; Ovalbumin; Rats; Th2 Cells

Drug delivery systems (DDS) are based on the concept of providing the optimal amount of a drug to a specific area requiring treatment. Liposomes are lipid-based nanoparticles capable of encapsulating any drug into both their membrane and aqueous phases. They have the potential to be targeted when their surfaces are modified with functional molecules such as antibodies and peptides. Thus, liposomes have strong potential as drug carriers if designed for active targeting. Our research group has recently developed a new concept for liposome targeting called "reverse targeting DDS (RT-DDS)". RT-DDS differs from conventional active targeting in that the surface of the liposomes is modified with an antigenic molecule that is specifically recognized by antigen-specific immune cells. This review describes in detail the differences between these two DDS targeting concepts and proposes the application of RT-DDS to the treatment of allergies based on research using ovalbumin as a model allergy antigen.

Topics: Animals; B-Lymphocytes; Disease Models, Animal; Drug Delivery Systems; Humans; Hypersensitivity; Immunosuppressive Agents; Liposomes; Mice; Ovalbumin; Spleen; Tacrolimus

The immune response against helminths and allergens is generally characterized by high levels of IgE and increased numbers of Th2 cells, eosinophils, and basophils. Basophils represent a relatively rare population of effector cells and their in vivo functions are incompletely understood. Recent studies with basophil-depleting antibodies revealed that these cells might play an important role during the early and late stages of type 2 immune responses. To further characterize the relevance of basophils for protective immunity and orchestration of allergic inflammation, we generated constitutively basophil-deficient mice. We observed a normal Th2 response induced by helminth infections or immunization with alum/OVA or papain/OVA. However, basophils contributed to worm expulsion during secondary helminth infection and mediated an IgE-dependent inflammatory response of the skin. These results argue against a critical role of basophils as antigen-presenting cells for induction of Th2 polarization and highlight their effector cell potential during later stages of a type 2 immune response.

Topics: Adaptive Immunity; Allergens; Animals; Basophils; Cytotoxicity, Immunologic; Helminthiasis; Humans; Hypersensitivity; Immunity, Innate; Immunoglobulin E; Inflammation; Mice; Mice, Transgenic; Ovalbumin; Th2 Cells

Allergic asthma is defined as a hypersensitivity reaction of the lung towards per se harmless antigens, e.g. pollen and house dust mite, accompanied with a chronic eosinophilic inflammation of the lung. During the course of the disease, physiological and structural changes in the lung occur, i.e. airway hyperresponsiveness, restricted airflow and airway remodelling. In addition to ovalbumin-induced mouse models of acute asthma, recently new models were developed, which show a closer resemblance to human asthma, both regarding the induction of characteristics of chronic allergic inflammation and the use of clinical relevant allergens. Moreover, attention is paid on the influence of adjuvants or the route of sensitisation on the protocol outcomes. The effort spent in development of these new models will be worthwhile, especially for research in the field of immuno-therapy. These improved animal models may broaden the knowledge of the disease and thereby provide new strategies for preventive and therapeutic interventions.

Topics: Allergens; Animals; Antigens, Plant; Asthma; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Mice; Ovalbumin; Plant Extracts

House dust mite (HDM) is the most pervasive indoor aeroallergen source worldwide. Allergens derived from HDM are associated with sensitization and allergic asthma. Allergic asthma is an immunologically driven disease characterized by a Th2-polarized immune response, eosinophilic inflammation, airway hyperreactivity, and remodeling. Animal models of asthma utilizing ovalbumin (OVA) exposure have afforded us considerable insight with respect to the mediators and cell types involved in allergic airway inflammation. However, OVA preparations and HDM are two vastly different materials. This chapter is specifically concerned with modeling responses to HDM exposure in mice. These studies have furnished new information and unlocked new lines of inquiry regarding biological responses to common aeroallergens. The complexity of HDM as an allergen source, with its plethora of protein and nonprotein immunogenic components, may influence the mechanisms underlying sensitization, inflammation and remodeling. Here, we will discuss this issue, along with giving critical thought to the use of experimental models.

Topics: Animals; Antigens, Dermatophagoides; Asthma; Bronchial Hyperreactivity; Disease Models, Animal; Humans; Hypersensitivity; Mice; Ovalbumin; Pyroglyphidae; Th2 Cells

Topics: Allergens; Animals; Biological Availability; Cytokines; Humans; Hypersensitivity; Immunization; Immunoglobulin E; Injections, Intraperitoneal; Interferon-gamma; Lymphoid Tissue; Nebulizers and Vaporizers; Ovalbumin; T-Lymphocytes; Th1 Cells; Th2 Cells

Topics: Adrenal Glands; Anaphylaxis; Animals; Antigen-Antibody Reactions; Carbohydrates; Egg White; Fibrinolysin; Globulins; Histamine; Hormones; Humans; Hypersensitivity; Kinins; Mast Cells; Muramidase; Neuraminic Acids; Ovalbumin; Pancreas; Rats; Serotonin; Thyroid Gland

Whether breast milk (BM) can protect against allergy has been studied extensively, with conflicting results. Variations in mothers' BM composition may explain some of the conflicting results. Our aim was to assess the impact of maternal allergy and probiotic intervention on BM food antibodies, transforming growth factor (TGF)-β(2) and interleukin (IL)-10 and their impact on allergy development in children until the ages of 2 and 5.. We measured total IgA, IgA antibodies to cow's milk (CM), casein, β-lactoglobulin and ovalbumin (OVA), TGF-β(2) and IL-10 in 364 colostrum samples and 321 BM samples taken at 3 months from mothers participating in a prospective study evaluating the allergy-preventive effect of probiotics in a cohort with an increased risk for allergy.. CM, casein and OVA antibodies, TGF-β(2) and IL-10 were detectable in most samples. Maternal allergy was associated with raised levels of IgA to casein (p = 0.04) and lower levels of TGF-β(2) (p = 0.006) in mature BM. Probiotic supplementation was associated with increased IL-10 (p = 0.046) and decreased casein IgA antibodies (p = 0.027) in mature BM. High OVA IgA antibodies in colostrum were associated with the development of atopy by the age of 2, while low levels in mature BM were a significant risk factor for the development of eczema by the age of 2. TGF-β(2) levels in BM constituted a risk for development of allergy by the age of 2.. The immunologic composition of BM was only slightly affected by maternal atopy and could be altered by probiotic supplementation. Small effects of BM components on allergy development in children were evident.

Topics: Animals; Caseins; Cattle; Child, Preschool; Colostrum; Cytokines; Double-Blind Method; Eczema; Female; Food; Humans; Hypersensitivity; Hypersensitivity, Immediate; Immunity, Maternally-Acquired; Immunoglobulin A; Infant; Infant, Newborn; Interleukin-10; Lactoglobulins; Male; Milk; Milk, Human; Ovalbumin; Pregnancy; Pregnancy Complications; Probiotics; Prospective Studies; Risk Factors; Transforming Growth Factor beta2

A reduction in microbial burden during infancy when allergen-specific memory is evolving has become a prominent explanation for the allergy epidemic.. We sought to determine whether probiotic dietary supplementation in the first 6 months of life could modify allergen- and vaccine-specific immune responses.. Two hundred and thirty-one pregnant women with a history of allergic disease and positive allergen skin prick test (SPT) were recruited into a randomized-controlled trial. The infants received either a probiotic (3 x 10(9)Lactobacillus acidophilus LAVRI-A1; Probiomics) or placebo (maltodextrin alone) daily for the first 6 months of life, given independent of feeding methods. One hundred and seventy-eight children completed the study; blood samples were available from 60 children in the placebo group and 58 children in the probiotic group. Infant cytokine (IL-5, IL-6, IL-10, IL-13, TNF-alpha or TGF-beta) responses to tetanus toxoid (TT), house dust mite (HDM), ovalbumin (OVA), beta-lactoglobulin (BLG), Staphylococcus enterotoxin B (SEB) and phytohaemaglutinin (PHA) were measured at 6 months of age.. Children who received the probiotics showed reduced production of IL-5 and TGF-beta in response to polyclonal (SEB) stimulation (P=0.044 and 0.015, respectively). They also demonstrated significantly lower IL-10 responses to TT vaccine antigen compared with the placebo group (P=0.03), and this was not due to any differences in vaccination. However, there were no significant effects of probiotics on either Type 1 (Th1) or Type 2 (Th2) T helper cell responses to allergens or other stimuli. The only other effects observed were for reduced TNF-alpha and IL-10 responsiveness to HDM allergens in children receiving probiotics (P=0.046 and 0.014, respectively).. In summary, although we did not see any consistent effects on allergen-specific responses, our study suggests that probiotics may have immunomodulatory effects on vaccine responses. The significance and clinical relevance of this need to be determined in further studies.

Topics: Allergens; Antigens, Dermatophagoides; Cells, Cultured; Dietary Supplements; Enterotoxins; Humans; Hypersensitivity; Infant; Infant, Newborn; Interleukin-10; Interleukin-5; Lactobacillus acidophilus; Lactoglobulins; Leukocytes, Mononuclear; Neutrophils; Ovalbumin; Phytohemagglutinins; Probiotics; Tetanus Toxoid; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha; Vaccines

Maternal avoidance of egg intake has been recommended to treat egg allergy in breastfed infants.. To determine if the concentration of ovalbumin (OVA) in human milk is directly related to the quantity and form of egg consumed by breastfeeding mothers.. Randomized, blinded, cross-over, intervention trial. Breastfeeding women (n = 41) attended four clinic days between 11 and 14 weeks of lactation and on each day were randomly allocated to receive a test breakfast, identical except for the egg content (no egg, one raw egg, half a cooked egg or one cooked egg). Breast milk samples were collected at two hourly intervals for 8 h and their OVA concentration measured by ELISA.. There was a direct, dose-response between the amount of cooked egg ingested and the peak OVA concentration (no egg 0.05 ng/mL [95% confidence interval (CI), 0.01-0.11], half a cooked egg 2.24 ng/mL [95% CI, 0.57-3.91], one cooked egg 3.16 ng/mL [95% CI, 1.41-4.91], n = 41, P<0.05) as well as the total OVA excretion (no egg 0.18 ng/mL/h [95% CI, 0.04-0.39], half a cooked egg 4.93 ng/mL/h [95% CI, 1.40-8.46], one cooked egg 9.14 ng/mL/h [95% CI, 4.25-14.03], n = 41, P<0.05). The peak concentration and total OVA excretion in response to one raw egg did not differ from ingesting half a cooked egg. There was no detectable OVA in the breast milk of 24% (10/41) women up to 8 h after any egg challenge.. OVA was detected in the breast milk of lactating women up to 8 h after a controlled intake of egg. A dose-response correlation was indicated. As excretion of OVA in human milk appears to be a normal phenomenon, further studies need to determine the threshold of OVA excretion that leads to symptoms in egg-allergic breastfed infants.

Topics: Adult; Animals; Breast Feeding; Chickens; Cross-Over Studies; Diet; Double-Blind Method; Egg Hypersensitivity; Eggs; Female; Food Handling; Humans; Hypersensitivity; Milk, Human; Ovalbumin

The value of allergen elimination diets during pregnancy for primary prevention of infant allergy has been questioned. However, dietary compliance may influence effectiveness.. To monitor egg intake during a randomized controlled trial of egg avoidance throughout pregnancy and lactation by serial measurements of serum ovalbumin (OVA) IgG concentration in conjunction with dietary diary record and also, to analyse specific IgG concentrations at birth in relation to infant allergic outcome.. Pregnant women, with personal or partner atopy, were randomized to complete dietary egg exclusion or an unmodified healthy diet before 20 weeks gestation. The infants were evaluated for atopy at 6 months of age. Serum food-specific IgG concentrations were determined by ELISA in maternal samples collected at study recruitment and during labour, and in infant samples at birth (umbilical cord).. Serum-specific IgG to OVA, but not the unrelated allergen, cow's milk beta-lactoglobulin, decreased over pregnancy in egg-avoiding women only (P<0.001). Cord OVA IgG concentration correlated with maternal IgG at delivery (r=0.944; P<0.001), and for infants born to atopic women, cord concentration was higher than that of their mother's (P<0.001). Infants with the lowest and highest cord IgG concentrations were the least likely, and those with mid-range concentrations were the most likely, to be atopic by 6 months of age (P=0.008).. Serum OVA IgG concentration reflects egg consumption, thereby indicating dietary allergen doses to which the developing immune system might be exposed. Trans-placental maternal IgG must be considered among early life factors that regulate infant atopic programming.

Topics: Adult; Animals; Chi-Square Distribution; Diet; Diet Records; Eggs; Enzyme-Linked Immunosorbent Assay; Female; Fetal Blood; Humans; Hypersensitivity; Immunoglobulin G; Infant; Infant, Newborn; Lactation; Ovalbumin; Patient Compliance; Pregnancy; Pregnancy Trimester, Second; Pregnancy Trimester, Third; Prospective Studies

The ingestion of food antigens usually results in the induction of oral tolerance, but the clinical and immunologic consequences of brief exposure to cow's milk proteins during the neonatal period are not well-documented. The aim of this work was to study immunoglobulin (Ig)E and IgG responses to cow's milk proteins and ovalbumin after exposure during the first three days of life in infants who were otherwise exclusively breast-fed. A group of 129 infants was randomly assigned at birth to one of three feeding regimens: human milk (HM), cow's milk formula (CMF), or a casein hydrolysate formula (CHF), during the first three days of life. They were then all exclusively breast-fed for a varying period of time and followed for two years. Serum IgG and IgE antibodies to cow's milk proteins and ovalbumin (OVA) were analyzed in blood samples obtained at birth, at 4 days and at 2, 4, 8, 12 and 24 months of age. The levels of IgG antibodies to beta-lactoglobulin (IgG-BLG) and bovine serum albumin (IgG-BSA) were higher in the CMF and the HM groups than in the CHF group for up to two years. This was particularly obvious for IgG-BLG in infants who started weaning before two months. The levels of IgG antibodies to casein (IgG-CAS) were higher in the CMF group, as compared with the CHF group at 8 and 12 months. The levels of IgG antibodies to OVA were similar in all three feeding groups. The levels of IgE antibodies to CAS or OVA were similar in the three feeding groups. Exposure to cow's milk during the first three days of life stimulated IgG antibody production to cow's milk proteins and this was still obvious at 2 years of age, while feeding with a casein hydrolysate during the first three days of life was associated with low levels of IgG antibodies to cow's milk proteins.

Topics: Animals; Antibody Specificity; Bottle Feeding; Caseins; Cattle; Child, Preschool; Female; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Infant; Infant, Newborn; Lactoglobulins; Longitudinal Studies; Male; Milk Proteins; Ovalbumin; Prevalence; Protein Hydrolysates; Serum Albumin, Bovine; Time Factors

Angelica decursiva Franchet & Savatier is a traditional medicinal plant used to treat asthma, cough, headache, pyrexia and thick phlegm in China, Japan and Korea. A. decursiva contains many types of coumarins, which can exert several pharmacological activities including anti-inflammatory and antioxidant properties for treating various diseases such as pneumonitis, atopic dermatitis, diabetes, and Alzheimer's disease.. In this study, we analyzed the components of A. decursiva ethanol extract (ADE) by high performance liquid chromatography (HPLC) and investigated the therapeutic effects of ADE against allergic asthma using lipopolysaccharide (LPS) stimulated RAW264.7 cells and an ovalbumin (OVA)-exposed allergic asthma model. To elucidate the mechanism of action of ADE, we examined the protein expression through network pharmacological analysis.. To establish asthma model, the mice were sensitized on day 0 and 14 via intraperitoneal injection of OVA with aluminum hydroxide. The mice were inhaled with OVA using an ultrasonic nebulizer on day 21, 22 and 23. ADE (50 and 100 mg/kg) was administered to mice by oral gave form day 18-23. On day 24, airway hyperresponsiveness (AHR) was measured using flexivent. On day 25, the mice were sacrificed and collected bronchoalveolar lavage fluids (BALF), serum and lung tissue. In LPS-stimulated RAW264.7 cell, nitric oxide and cytokines were measured. Additionally, expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-κB were detected using double-immunofluorescence.. We detected the five coumarin components which included nodakenin, umbelliferon, (-)-marmesin (=nodakenetin), bergapten, and decursin, in ADE by high performance liquid chromatography. Treatment with ADE decreased the production of nitric oxide, interleukin (IL)-6 and tumor necrosis factor (TNF)-α in LPS-stimulated RAW264.7 cells accompanied by the enhanced expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-κB. In the asthma model, the administration of ADE reduced inflammatory cell count and airway hyperresponsiveness in OVA-exposed animals with decreased levels of IL-4, IL-13, and OVA-specific immunoglobulin E. These results were accompanied by the reduction of pulmonary inflammation and mucus secretion. Furthermore, ADE administration inhibited the expression of NF-κB and matrix metalloproteinase (MMP)-9 in OVA-exposed animals, which was consistent with the results of network pharmacological analysis.. This study demonstrated that ADE effectively attenuated allergic inflammation induced by OVA inhalation through the enhancement of Nrf2 expression and suppression of NF-κB expression. Therefore, ADE may be a potential therapeutic agent for controlling asthma.

Topics: Angelica; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hypersensitivity; Interleukin-6; Lipopolysaccharides; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; NF-kappa B; Nitric Oxide; Ovalbumin; Pneumonia

Eclipta prostrata (L.) L. is a medicinal plant used by many ethnic groups in Brazil to treat respiratory diseases, hepatitis and the bites of venomous animals. A methanolic extract of E. prostrata (MEEP), the major components of which are wedelolactone (WED) and demethylwedelolactone (DMW), exhibited anti-inflammatory activity in acute asthma models but the effects on lung inflammation and the mechanisms of action of MEEP in a chronic asthma model are not known.. To study the effects of MEEP in vivo using a chronic ovalbumin (OVA)-induced allergic asthma model in mice.. The identities of WED and DMW in MEEP were confirmed and the concentrations determined by liquid chromatography and tandem mass spectrometry. Male Balb/c mice were sensitized and challenged with OVA and experimental animals were treated with MEEP (100, 250 or 500 mg/kg) while control animals were treated with dexamethasone (2 mg/kg) or normal saline. Bronchial hyperresponsiveness, total and differential cell counts in bronchoalveolar lavage (BAL), and the production of Th2 cytokines in lung homogenates were assessed. Lung inflammation and mucus production were evaluated by histological analysis while nuclear factor kappa-B (NF-κB) activation was assessed immunohistochemically.. Concentrations of WED and DMW in MEEP were 5.12% and 1.04%, respectively. Treatments with MEEP (250 or 500 mg/kg) significantly decreased bronchial hyperresponsiveness, reduced total cell and eosinophil counts in BAL and IL-4 concentrations in lung homogenate, and inhibited NF-κB activation. Treatment with MEEP at 500 mg/kg reduced the level of IL-5 in lung homogenates but did not decrease IL-13 concentration or mucus production.. MEEP attenuated bronchial hyperresponsiveness and decreased lung and airway inflammation in a chronic asthma model in mice. The mechanism of action involves inhibition of NF-κB activation, most likely associated with the presence of the coumestans WED and DMW. These results support the ethnopharmacological evidence for the use of E. prostrata against asthma and other respiratory inflammatory diseases.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eclipta; Hypersensitivity; Inflammation; Lung; Methanol; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

To date, few studies have been conducted on the relationship between postbiotics and air pollution, and there is limited knowledge if postbiotic and probiotic have synergistic effects. Therefore, we created a PM-induced lung inflammation mice model and demonstrated the effect of probiotic, postbiotic, and their combination treatment on attenuation of PM2.5-induced lung damage and allergic response. The mice were intratracheally given PM2.5 triggering conditions of acute lung damage and allergic response. Our results showed that individual treatment of probiotic and postbiotic reduced body weight loss by 47.1% and 48.9%, but the results did not show any effect on polarizing IFN-γ/IL-4 ratio. In addition, PM2.5-induced overactive expression of IgE treated by probiotic and postbiotic was reduced by 33.2% and 30.4%, respectively. While combination treatment of probiotic and postbiotic exerted a synergistic effect, especially considerably on improving IgE reduction by 57.1%, body weight loss by 78.3%, and IFN-γ/IL-4 ratio boost by 87.5%. To sum up the above functionality, these research findings may help establish a novel platform for postbiotic application, formulation, and mechanistic selection with regard to PM2.5-induced lung injury. PRACTICAL APPLICATION: Allergic inflammation caused by PM2.5 is not like common allergens (ex. Pollens, ovalbumin, dust mites), which simply skewing Th1/Th2 polarization to Th2. Thus using probiotics screened by Th1-skewing criteria might not be the best choice to treat on PM2.5-induced symptoms. This research proposed a combination of probiotics and postbiotics on modulating immunity homeostasis, and consequently attenuating complications of PM2.5-induced lung damage. These research findings may help establish a novel platform for postbiotic application, formulation and mechanistic selection with regard to PM2.5-induced lung injury.

Topics: Animals; Cytokines; Hypersensitivity; Immunoglobulin E; Interleukin-4; Lung; Lung Injury; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Pneumonia; Probiotics; Weight Loss

Translationally controlled tumor protein (TCTP), a highly conserved protein present in most eukaryotes, is involved in numerous biological processes. Only the dimeric form of TCTP (dTCTP) formed during inflammatory conditions exhibits cytokine-like activity. Therefore, dTCTP is considered as a therapeutic target for allergic diseases. Because monomeric TCTP (mTCTP) and dTCTP share a high topological similarity, we hypothesized that small molecules interacting with mTCTP would also bind to dTCTP and interfere with dTCTP-based cellular processes. In this study, nine compounds listed in the literature as interacting with mTCTP were investigated for their ability to suppress the activity of extracellular dTCTP in bronchial epithelial cells. It was found that one of the nine, meclizine, a piperazine-derivative antihistamine, significantly reduced IL-8 release and suppressed the NF-κB pathway. The direct interaction of meclizine with dTCTP was confirmed by surface plasmon resonance (SPR). Also, we found that meclizine can attenuate ovalbumin (OVA)-induced airway inflammation in mice. Therefore, meclizine might be a potential anti-allergic drug as an inhibitor for dTCTP.

Topics: Animals; Biomarkers, Tumor; Disease Models, Animal; Histamine Antagonists; Hypersensitivity; Meclizine; Mice; Mice, Inbred BALB C; Ovalbumin; Piperazine; Tumor Protein, Translationally-Controlled 1

The plasticizer di- (2-ethylhexyl) phthalate (DEHP) is considered a risk factor for allergic diseases and has attracted public attention for its adverse effects on health. However, respiratory adverse effects after DEHP exposure in food allergies have rarely been reported. MiRNAs are considered to be key regulators in the complex interrelationships between the host and microbiome and may be a potential factor involved in DEHP-induced pulmonary toxicity. To investigate the adverse effects of DEHP on the lung during sensitization, we established an ovalbumin (OVA)-sensitized mouse model exposed to DEHP and performed 16S rDNA gene sequencing, miRNA sequencing, and correlation analysis. Our results showed that DEHP aggravated the immune disorder in OVA-sensitized mice, which was mainly characterized by an increase in the proportion of Th2 lymphocytes, and further enhanced OVA-induced airway inflammation without promoting pulmonary fibrosis. Compared with the OVA group, DEHP interfered with the lung microbial community, making Proteobacteria the dominant phylum, while Bacteroidetes were significantly reduced. Differentially expressed miRNAs were enriched in the PI3K/AKT pathway, which was closely related to immune function and airway inflammation. The expression of miR-146b-5p was elevated in the DEHP group, which was positively correlated with the proportion of Th2 cells and significantly negatively correlated with the abundance of Bacteroidetes. The results indicate that DEHP may interfere with the expression of miR-146b-5p, affect the composition of the lung microbiota, induce an imbalance in T cells, and lead to immune disorders and airway inflammation. The current study uses multi-omics to reveal the potential link between the plasticizer DEHP and allergic diseases and provides new insights into the ecotoxicology of environmental exposures to DEHP.

Topics: Animals; Diethylhexyl Phthalate; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Multiomics; Ovalbumin; Phosphatidylinositol 3-Kinases; Plasticizers

Asthma is a chronic airway disease. Allergic reactions and T helper (h)2 immune response play a key role in asthma occurrence. Cell therapy can control inflammation and remodeling responses in allergic asthma, and cytokines can change this effect. Therefore, in this study, the effect of treated cell therapy with IL-2 to control allergic asthma was studied. Bone marrow cells were extracted and co-cultured with IL-2 and the cells were used via intra-tracheal administration in allergic asthma mice. Levels of IL-4, IL-5, IL-13, Leukotriene B4 and C4, and remodeling factors were measured. At least, a histopathology test of lung tissue was done. Type2 cytokines, leukotrienes, remodeling factors, mucus secretion, goblet cell hyperplasia, peri-bronchial and peri-vascular inflammation were significantly (p˂0.05) decreased by treating with bone marrow-derived mononuclear cells (BMDMCs) and IL-2-BMDMCs. Treatment with IL-2-BMDMCs could significantly decrease IL-13, transforming growth factor (TGF)-β, HP levels, and mucus secretion (p˂0.05) compared to BMDMCs treatment. In this study, BMDMCs and IL-2-BMDMCs therapy could decrease inflammation, allergic, and remodeling factors in allergic asthma. Cell therapy with BMDMCs had a strong and notable effect on the control of allergic asthma pathophysiology when co-cultured and used with IL-2.

Topics: Animals; Asthma; Bone Marrow; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Inflammation; Interleukin-13; Interleukin-2; Leukocytes, Mononuclear; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta

Topics: Animals; Cell Degranulation; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Interleukin-13; Interleukin-4; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphatidylinositol 3-Kinases

Topics: Administration, Topical; Animals; Cytokines; Gardenia; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin

Dibutyl phthalate (DBP), used as a plasticizer, is of wide concern as an environmental pollutant since it has certain immunotoxicity. Although there is growing evidence supporting a link between DBP exposure and allergic airway inflammation, there is less information concerned with whether the ferroptosis pathway is involved in DBP-aggravated allergic asthma in ovalbumin (OVA)-sensitized mice. This study aimed to investigate the role and underlying mechanisms of ferroptosis in DBP-exposed allergic asthmatic mice. Balb/c mice were orally exposed to 40 mg/kg

Topics: Animals; Asthma; Dibutyl Phthalate; Ferroptosis; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Reactive Oxygen Species

The currently observed high prevalence of allergic diseases has been associated with changes in microbial exposure in industrialized countries. Defined bacterial components represent a new strategy for modulating the allergic immune response. We show that intranasal administration of exopolysaccharide (EPS) isolated from Lacticaseibacillus (L.) rhamnosus LOCK900 induces TGF-β1, IgA, and regulatory FoxP3

Topics: Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hypersensitivity; Immunoglobulin A; Inflammation; Lacticaseibacillus; Lacticaseibacillus rhamnosus; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Allergies are increasing worldwide. The presence of atopic diseases in the mother propagates the onset of allergic diseases in the offspring with a considerably stronger penetrance than atopic diseases of the father. Such observation challenges genetic predispositions as the sole cause of allergic diseases. Epidemiological studies suggest that caregiver stress in the perinatal period may predispose offspring to asthma. Only one group has studied the link between prenatal stress and neonatal asthma susceptibility in a murine model.. We aimed to study if the neonatal increased risk of developing allergic lung inflammation persists after puberty and if there are sex differences in susceptibility.. Pregnant BALB/c mice were subjected to a single restraint stress exposure at day 15 of gestation. Pups were separated by gender and subjected to a well-known sub-optimal asthma model after puberty.. Adult mice born to stressed dams were more susceptible to developing allergic pulmonary inflammation since an increase in the number of eosinophils in bronchoalveolar lavage (BAL), a greater peribronchial and perivascular infiltrate, a higher proportion of mucus-producing cells, and increased IL-4 and IL-5 levels in BAL were detected compared to control mice. These effects were more profound in females than males. Moreover, only females from stressed dams showed an increase in IgE levels.. Increased litter susceptibility to develop allergic lung inflammation induced by maternal stress persists after puberty and is more potent in females than in male mice.

Topics: Animals; Asthma; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Pregnancy

Exposure to environmental pollutants, including benzo[a]pyrene (BaP), has been implicated in allergic diseases and intestinal microbiota homeostasis, but the environment-microbiota-immunity triangular relationship and to what extent BaP-induced remodeling of the gut microbiota contributes to intestinal allergic inflammation remain to be established.. We investigated the impact of BaP on intestinal allergic inflammation and examined the relationship between this effect and gut microbiota dysbiosis. We explored the potential ability of intestinal bacteria to degrade BaP and alleviate cytotoxicity as a detoxification strategy to counteract the effects of BaP exposure.. We combined microbiome shotgun metagenomics with animal histological and intestinal allergic inflammatory responses to assess the effects of BaP (. BaP exposure impacted the taxonomic composition and the functional potential of the gut microbiota and aggravated antigen-induced intestinal allergic inflammatory responses. The level of inflammatory cytokines correlated with the abundance of specific bacterial taxa, including. Using allergic female mice as a model, we investigated the relationship between BaP, microbiota, and host immune reactions, highlighting the role of gut bacteria in BaP-aggravated allergic reactions. Our findings offer novel insight toward establishing the causal relationship between BaP exposure and the occurrence of allergic disorders. Identifying gut bacteria that degrade BaP may provide new strategies for ameliorating BaP cytotoxicity. https://doi.org/10.1289/EHP11874.

Topics: Animals; Bacteria; Caco-2 Cells; Female; Gastrointestinal Microbiome; Humans; Hypersensitivity; Inflammation; Mice; Ovalbumin

Docosahexaenoic acid (DHA) and arachidonic acid (AA) on oral tolerance (OT) development in allergy-prone infants is less known.. We aim to determine the effects of early life DHA supplementation (1% of total fat, from novel canola oil), along with AA, on OT toward ovalbumin (ova, egg protein) in allergy-prone BALB/c pups at 6-wk.. Breastfeeding dams (n ≥ 10/diet) were fed DHA+AA (1% DHA, 1% AA wt/wt of total fat) or control (0% DHA, 0% AA) suckling period diet (SPD) during which pups consumed dam's milk. At 3-wk, pups from each SPD group were assigned to either the control or DHA+AA weaning diet. For OT, pups from each diet group were either orally fed ova or placebo daily from 21-25 d. Systemic immunization to ova was induced through intraperitoneal injections before euthanizing 6-wk pups. Ova-specific immunoglobulin (ova-Ig) and splenocytes ex-vivo cytokine response to different stimuli were analyzed using a 3-factor analysis of variance.. OT-induced suppression was seen in ova-stimulated splenocyte ex-vivo response, where ova-tolerized pups showed significantly lower total immunoglobulin (Ig)G, IgG1, interleukin (IL)-2 and IL-6 production than sucrose (placebo) pups. DHA+AA SPD was associated with 3 times lower plasma concentrations of ova-IgE (P = 0.03) than controls. DHA+AA weaning diet resulted in lower T helper type-2 cytokines (IL-4 and IL-6) with ova stimulation than controls, which may benefit OT. DHA+AA SPD resulted in significantly higher T cell cytokine response [IL-2, interferon-gamma, (IFNγ) and IL-1β] to anti-CD3/CD28 stimulation than controls. The splenocytes stimulated with lipopolysaccharide produced lower inflammatory cytokines (IFNγ, tumor necrosis factor-alpha, IL-6, and C-X-C motif ligand 1), which may be because of lower CD11b+CD68+ splenocytes proportion in pups from DHA+AA SPD than control (all P < 0.05).. DHA and AA in early life may influence OT in allergy-prone BALB/c mouse offspring, as they effectively promote T helper type-1 immune responses.

Topics: Animals; Arachidonic Acid; Cytokines; Docosahexaenoic Acids; Hypersensitivity; Immune Tolerance; Immunoglobulin G; Interleukin-6; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes

There is now sufficient evidence to support an inverse association between helminth infection and secreted products with allergic/autoimmune disorders. Accordingly, several experimental studies have shown that Echinococcus granulosus infection and hydatid cyst compounds are able to suppress immune responses in allergic airway inflammation. This is the first study on effects of somatic antigens of E. granulosus on chronic allergic airway inflammation in BALB/c mice. Mice in OVA group were intraperitoneally (IP) sensitized with OVA/Alum. Subsequently, were challenged by nebulizing of OVA 1%. The treatment groups received somatic antigens of protoscoleces on the specified days. Mice in PBS group were received PBS in both sensitization and challenge. The effects of somatic products on development of chronic allergic airway inflammation were evaluated by examining histopathological changes, the recruitment of inflammatory cells in the bronchoalveolar lavage, cytokines production in the homogenized lung tissue, and total antioxidant capacity in serum. Our findings show that the co-administration of somatic antigens of protoscoleces simultaneously with the development of asthma intensifies allergic airway inflammation. The identification of effective components involved in exacerbation of allergic airway inflammation manifestations will be a crucial approach to understanding the mechanism of these interactions.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Disease Progression; Echinococcosis; Echinococcus granulosus; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is a chronic inflammatory disease of the airways with a high prevalence. Asthma has a complex pathophysiology and about 5-10% of patients are not fully responsive to the currently available treatments. The aim of this study is to investigate the involvement of NF-κB in the effects of fenofibrate on a mouse model of allergic asthma.. A total of 49 BALB/c mice were randomly distributed into 7 groups (n = 7). Allergic asthma model was created by administering i.p. injections of ovalbumin on days 0, 14 and 21, followed by provocation with inhaled ovalbumin on days 28, 29 and 30. Fenofibrate was orally given in 3 different doses; 1, 10 and 30 mg/kg through days 21-30 of the experiment. On day 31, pulmonary function test using whole body plethysmography was performed. The mice were sacrificed 24 h later. Blood samples were obtained, and serum of each sample was separated for IgE determination. Bronchoalveolar lavage fluid (BALF) and lung tissues were collected to measure IL-5 and IL-13 levels. Nuclear extracts of lung tissues were employed to assess nuclear factor kappa B (NF-κB) p65 binding activity.. Enhanced Pause (Penh) values were significantly increased in ovalbumin-sensitized and challenged mice (p < 0.01). Administration of fenofibrate (10 and 30 mg/kg) resulted in improved pulmonary function as shown by significantly lower Penh values (p < 0.01). Interleukin (IL) - 5 and IL-13 levels in BALF and lung tissues and immunoglobulin E (IgE) levels in serum were significantly elevated in the allergic mice. IL-5 levels in the lung tissues of mice treated with 1 mg/kg fenofibrate (FEN1) group were significantly reduced (p < 0.01). BALF and lung tissue IL-5 and IL-13 levels in mice treated with 10 and 30 mg/kg fenofibrate, FEN10 and FEN30, respectively, were significantly diminished when compared to the ovalbumin-treated (OVA) group, whereas treatment with 1 mg/kg fenofibrate resulted in insignificant changes. IgE levels in the serum of FEN30 group mice have shown a prominent reduction (p < 0.01). NF-κB p65 binding activity was higher in mice sensitized and challenged with ovalbumin (p < 0.01). NF-κB p65 binding activity was significantly reduced in allergic mice treated with 30 mg/kg (p < 0.01) fenofibrate.. In this study, we showed that administration of 10 and 30 mg/kg fenofibrate effectively attenuated airway hyperresponsiveness and inflammation in a mouse model of allergic asthma, possibly through inhibition of NF-κB binding activity.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Fenofibrate; Hypersensitivity; Immunoglobulin E; Interleukin-13; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Previous studies have reported a correlation between the dysregulation of intestinal microbiota and the occurrence of asthma. This study aimed to investigate the effect of probiotic Lactobacillus rhamnosus 76 (LR76) on ovalbumin (OVA)-allergic mice and the mechanism of LR76 affecting mucus secretion in asthma. OVA-allergic mice were supplemented with LR76, and 16HBE cells induced by interleukin-13 (IL-13) were treated with LR76 supernatant (LR76-s) to observe the effect of LR76. In OVA-sensitized mice, LR76 alleviated the inflammatory cell infiltration in lung tissue and reduced the inflammatory cell counts of BALF. The expression level of mRNA, including Il4, Il5, Il13, Il25, Tgfb1, Il10, and Ifng, was decreased in the lung tissue of mice in the LR76 group compared with the OVA group. MUC5AC expression was down-regulated, while SCGB1A1 was up-regulated in the lung tissue of OVA-allergic mice after being supplemented with LR76 and in 16HBE cells induced by IL-13 after incubating with LR76-s. LR76 and LR76-s down-regulated the expression of proteins, including STAT6, p-STAT6, and SPDEF, and mRNA of STAT6 and SPDEF. In conclusion, LR76 alleviated airway inflammation and Th2 response in OVA-allergic mice and improved the mucus secretion of mouse lung tissue and 16HBE cells in the asthma model by down-regulating STAT6/SPDEF pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Hypersensitivity; Inflammation; Interleukin-13; Lacticaseibacillus rhamnosus; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; RNA, Messenger; Transcription Factors

Asthma is a chronic inflammatory lung disease that causes respiratory difficulties. Black ginseng extract (BGE) has preventative effects on respiratory inflammatory diseases such as asthma. However, the pharmacological mechanisms behind the anti-asthmatic activity of BGE remain unknown. To investigate the anti-asthmatic mechanism of BGE, phorbol 12-myristate 13-acetate plus ionomycin (PMA/Iono)-stimulated mouse EL4 cells and ovalbumin (OVA)-induced mice with allergic airway inflammation were used. Immune cells (eosinophils/macrophages), interleukin (IL)-4, -5, -13, and serum immunoglobulin E (IgE) levels were measured using an enzyme-linked immunosorbent assay. Inflammatory cell recruitment and mucus secretion in the lung tissue were estimated. Protein expression was analyzed via Western blotting, including that of inducible nitric oxide synthase (iNOS) and the activation of protein kinase C theta (PKCθ) and its downstream signaling molecules. BGE decreased T helper (Th)2 cytokines, serum IgE, mucus secretion, and iNOS expression in mice with allergic airway inflammation, thereby providing a protective effect. Moreover, BGE and its major ginsenosides inhibited the production of Th2 cytokines in PMA/Iono-stimulated EL4 cells. In EL4 cells, these outcomes were accompanied by the inactivation of PKCθ and its downstream transcription factors, such as nuclear factor of activated T cells (NFAT), nuclear factor kappa B (NF-κB), activator of transcription 6 (STAT6), and GATA binding protein 3 (GATA3), which are involved in allergic airway inflammation. BGE also inhibited the activation of PKCθ and the abovementioned transcriptional factors in the lung tissue of mice with allergic airway inflammation. These results highlight the potential of BGE as a useful therapeutic and preventative agent for allergic airway inflammatory diseases such as allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Panax; Signal Transduction

Secretion of translationally controlled tumor protein (TCTP) was found in body fluids during the late phase of allergic reactions, implicating TCTP in allergic diseases. Furthermore, blocking TCTP has been shown to be helpful in treating asthma and allergies in animal models. The objectives of this study were to produce anti-TCTP monoclonal antibodies (mAbs), test their ability to inhibit the cytokine-like function of dimeric TCTP (dTCTP) in vitro and to assess their therapeutic effects in a murine model of ovalbumin (OVA)-induced airway inflammation. We first verified the inhibitory effects of 4 anti-TCTP mAbs on dTCTP-induced secretion of IL-8 in BEAS-2B cells. To investigate the anti-inflammatory effect of anti-TCTP mAbs on allergic airway inflammation, we treated OVA-sensitized mice with anti-TCTP mAbs before OVA challenge. The changes in bronchoalveolar lavage fluid (BALF) cells, IL-4, IL-5, and IL-13 levels in both BALF and lung homogenates, plasma levels of OVA-specific IgE, and lung tissues were analyzed. We found that JEW-M449 anti-TCTP mAb bound to the flexible loop of TCTP and significantly inhibited dTCTP-induced IL-8 release, making it the most effective inhibitor in our study. We also found that treatment with JEW-M449 significantly reduced the infiltration of inflammatory cells and suppressed the OVA-induced upregulation of type 2 cytokines in both BALF and lung homogenates in a dose-dependent manner. In addition, JEW-M449 significantly attenuated the degree of goblet cell hyperplasia and mucus secretion. Our results demonstrate that specific targeting of the flexible loop of TCTP is a potent strategy for treating airway inflammatory diseases.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Inflammation; Interleukin-8; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Tumor Protein, Translationally-Controlled 1

Allergen immunotherapy (AIT) is the only curative treatment for allergic diseases. However, AIT has many disadvantages related to efficiency, safety, long-term duration, and patient compliance. Dendritic cells (DCs) have an important role in antigen-specific tolerance induction; thus, DC-targeting strategies to treat allergies such as glutaraldehyde crosslinked antigen to mannoprotein (MAN) have been established. However, glutaraldehyde crosslinking may reduce the antigen presentation efficiency of DCs. To overcome this, we developed a MAN-coated ovalbumin (OVA) nanoparticle (MDO), which uses intermolecular disulfide bond to crosslink OVA and MAN. MDO effectively targeted DCs resulting in tolerogenic DCs, and promoted higher antigen presentation efficiency by DCs compared with OVA or glutaraldehyde crosslinked nanoparticles. In vitro and in vivo experiments showed that DCs exposed to MDO induced T

Topics: Allergens; Animals; Dendritic Cells; Desensitization, Immunologic; Glutaral; Humans; Hypersensitivity; Immunotherapy; Mannans; Mice; Nanoparticles; Ovalbumin

The concept of innate and adaptive effector cells that are repleted by maturing inert progenitor cell populations is changing. Mast cells develop from rare mast cell progenitors populating peripheral tissues at homeostatic conditions, or as a result of induced recruitment during inflammatory conditions.. Because FcεRI-expressing mast cell progenitors are the dominating mast cell type during acute allergic lung inflammation in vivo, we hypothesized that they are activated by IgE cross-linking.. Mouse peritoneal and human peripheral blood cells were sensitized and stimulated with antigen, or stimulated with anti-IgE, and the mast cell progenitor population analyzed for signs of activation by flow cytometry. Isolated peritoneal mast cell progenitors were studied before and after anti-IgE stimulation at single-cell level by time-lapse fluorescence microscopy. Lung mast cell progenitors were analyzed for their ability to produce IL-13 by intracellular flow cytometry in a mouse model of ovalbumin-induced allergic airway inflammation.. Sensitized mouse peritoneal mast cell progenitors demonstrate increased levels of phosphorylation of tyrosines on intracellular proteins (total tyrosine phosphorylation), and spleen tyrosine kinase (Syk) phosphorylation after antigen exposure. Anti-IgE induced cell surface-associated lysomal-associated membrane protein-1 (LAMP-1) in naive mast cell progenitors, and prompted loss of fluorescence signal and altered morphology of isolated cells loaded with lysotracker. In human mast cell progenitors, anti-IgE increased total tyrosine phosphorylation, cell surface-associated LAMP-1, and CD63. Lung mast cell progenitors from mice with ovalbumin-induced allergic airway inflammation produce IL-13.. Mast cell progenitors become activated by IgE cross-linking and may contribute to the pathology associated with acute allergic airway inflammation.

Topics: Animals; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-13; Mast Cells; Mice; Ovalbumin; Receptors, IgE; Tyrosine

Mechanisms linking ingested pollutants to increased incidence of allergy are poorly understood. We report that mice exposed to low doses of cadmium develop higher IgE responses following oral allergen sensitization and more severe allergic symptoms upon allergen challenge. The environmentally relevant doses of this pollutant also induced oxidative/inflammatory responses in the gut of SPF, but not germ-free mice. Interestingly, the increased IgE responses correlated with stimulation of the vitamin D

Topics: 25-Hydroxyvitamin D3 1-alpha-Hydroxylase; Allergens; Animals; Cadmium; Disease Models, Animal; Environmental Pollutants; Humans; Hypersensitivity; Immunization; Immunoglobulin E; Intestines; Mice; Mice, Inbred C57BL; Ovalbumin; Signal Transduction; Vitamin D; Vitamin D3 24-Hydroxylase

Although exposure to ambient particulate matter (PM) is linked to asthma, the health effects of co-existing vapor-phase organic pollutants (vapor) and their combined effects with PM on this disease are poorly understood. We used a murine asthma model to test the hypothesis that exposure to vapor would enhance allergic sensitization and this effect would be further strengthened by co-existing PM. We found that vapor and PM each individually exerted adjuvant effects on OVA sensitization. Co-exposure to vapor and PM during sensitization further enhanced allergic lung inflammation and OVA-specific antibody production which was accompanied by pulmonary cytokine/chemokine milieu that favored T-helper 2 immunity (i.e. increased IL-4, downregulation of Il12a and Ifng, and upregulation of Ccl11 and Ccl8). TNFα, IL-6, Ccl12, Cxcl1 and detoxification/antioxidant enzyme responses in the lung were pollutant-dependent. Inhibition of lipopolysaccharide-induced IL-12 secretion from primary antigen-presenting dendritic cells correlated positively with vapor's oxidant potential. In conclusion, concurrent exposure to vapor and PM led to significantly exaggerated adjuvant effects on allergic lung inflammation which were more potent than that of each pollutant type alone. These findings suggest that the effects of multi-component air pollution on asthma may be significantly underestimated if research only focuses on a single air pollutant (e.g., PM).

Topics: Animals; Asthma; Cytokines; Down-Regulation; Drug Interactions; Female; Gene Expression Regulation; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Particulate Matter; RNA, Messenger; Th2 Cells; Up-Regulation; Volatile Organic Compounds

Topics: Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Antioxidants; Cell Degranulation; Colitis; Enteritis; Hypersensitivity; Interleukin-10; Male; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Resveratrol

Mounting evidence suggests that nematode infection can protect against disorders of immune dysregulation. Administration of live parasites or their excretory/secretory (ES) products has shown therapeutic effects across a wide range of animal models for immune disorders, including asthma. Human clinical trials of live parasite ingestion for the treatment of immune disorders have produced promising results, yet concerns persist regarding the ingestion of pathogenic organisms and the immunogenicity of protein components. Despite extensive efforts to define the active components of ES products, no small molecules with immune regulatory activity have been identified from nematodes. Here we show that an evolutionarily conserved family of nematode pheromones called ascarosides strongly modulates the pulmonary immune response and reduces asthma severity in mice. Screening the inhibitory effects of ascarosides produced by animal-parasitic nematodes on the development of asthma in an ovalbumin (OVA) murine model, we found that administration of nanogram quantities of ascr#7 prevented the development of lung eosinophilia, goblet cell metaplasia, and airway hyperreactivity. Ascr#7 suppressed the production of IL-33 from lung epithelial cells and reduced the number of memory-type pathogenic Th2 cells and ILC2s in the lung, both key drivers of the pathology of asthma. Our findings suggest that the mammalian immune system recognizes ascarosides as an evolutionarily conserved molecular signature of parasitic nematodes. The identification of a nematode-produced small molecule underlying the well-documented immunomodulatory effects of ES products may enable the development of treatment strategies for allergic diseases.

Topics: Animals; Asthma; Disease Models, Animal; Host-Pathogen Interactions; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Nematoda; Ovalbumin; Small Molecule Libraries; Trachea

Asthma is a common chronic airway inflammatory disease, lacking effective therapeutic approaches. Magnesium isoglycyrrhizinate (MgIG) is an anti-inflammatory drug for treating chronic inflammation. However, it is still ambiguous whether MgIG can function in allergy induced asthma. In this study, we investigated the anti-inflammation effect of MgIG in mice with allergy induced asthma and explored the underlying mechanisms.. Mouse asthma model was established with ovalbumin (OVA) sensitization and challenge. Subsequently, mice sensitized with OVA were randomly assigned into fourgroups: asthma model group (MDL), dexamethasone group (DXM), MgIG group (MgIG), and normal mice were used as normal control (CON). The mice in MgIG, MDL were given 0.2 mg/mL MgIG solution by atomization inhalation for 30 min before 1% (w/v) OVA challenge. At the completion of model establishment and drug treatment, cells in bronchoalveolar lavage fluid were classified, inflammatory factors in serum were determined, histopathological analysis was performed by H&E staining, and expression of MUC5AC, NLRP3, and cleaved caspase-1 in the lung tissue was also determined by immunohistochemistry and western blotting, respectively.. In comparison to MDL group, MgIG treatment could significantly inhibit the recruitment of white blood cells, neutrophils, and eosinophils in BALF, reduced the production of IL-6, TNF-α, and IgE in serum, and reduced mucus secretion and the infiltration of inflammatory cells. Also, an increase of NLRP3 and Caspase-1 protein levels were suppressed by MgIG treatment.. Our study findings support that nebulizer inhalation of MgIG as an effective therapy in treating the allergy induced asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Caspases; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Saponins; Triterpenes

Peptide-based immunotherapy (PIT) was introduced as an attractive approach in allergen-specific immunotherapy (AIT). However, PIT clinical trials have shown variable results, and immune response to peptides is not precisely predictable. On the other hand, induction of antigen-specific tolerance may be augmented when allergens are combined with the regulatory T cell epitope (Tregitope). This study aimed to evaluate the therapeutic administration of a plasmid DNA encoding Tregitope and ovalbumin (OVA) immunodominant epitope in the murine model of allergy.. Following the induction of allergic rhinitis by ovalbumin, vaccinated group received three doses of recombinant plasmid containing Signal peptide-Tregitope-OVA T cell epitope. After the final OVA challenge, clinical symptoms, histopathological changes, OVA-specific IgE level, and cytokine secretion pattern of spleen cells were examined.. Our data are showing that AIT with the recombinant DNA vaccine significantly suppressed airway inflammation; reduced eosinophilic infiltration in the nasal mucosa; decreased expression level of IL-4 and IL-17 in spleen cells, while IFN-γ, IL-10, and TGF-β expression were increased. Moreover, OVA-specific IgE levels were also decreased.. These results suggest that Tregitope-immunodominant T cell epitope fusion can act as a safe and effective approach in DNA-based allergen-specific immunotherapy.

Topics: Allergens; Animals; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Epitopes, T-Lymphocyte; Hypersensitivity; Immunodominant Epitopes; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Peptides; Plasmids

To clarify the detailed molecular mechanisms underlying the development of asthma, we assessed the potential immune effects of prenatal osteoprotegerin (OPG) inhibition in the pathogenesis of asthma. The effects of OPG deficiency on the development of asthma were evaluated using an ovalbumin-induced asthma model in OPG knockout mice. Histological analysis demonstrated that OPG was mainly detected in airway epithelial cells in wild type mice. After ovalbumin sensitization and challenge, accumulation of inflammatory cells, gene expression of T helper 2-related cytokines and mucus hypersecretion in lung tissues were inhibited by OPG deficiency. Importantly, the serum level of IgE was not increased in OPG KO mice after ovalbumin sensitization and challenge. Based on these findings, OPG knockout mice were protected against methacholine-induced airway hyperresponsiveness. OPG expression is thought to be essential for induction of the allergic immune response in asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Osteoprotegerin; Ovalbumin

Renifolin F is a prenylated chalcone isolated from

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Chalcone; Cytokines; Disease Models, Animal; Hypersensitivity; Immunity, Innate; Inflammation; Lung; Lymphocytes; Mice; MicroRNAs; Ovalbumin

Maternal improper nutrition has been reported to trigger respiratory disorders in offspring. Here, we characterized the effects of high-fat environment in the fetal period on mice and human cord blood CD4

Topics: Animals; Asthma; CD4-Positive T-Lymphocytes; Cell Differentiation; Cytokines; Diet, High-Fat; Disease Models, Animal; Hypersensitivity; Mice; Ovalbumin

The prevalence of allergic diseases is on the rise, yet the environmental factors that contribute to this increase are still being elucidated. Laundry detergent (LD) that contains cytotoxic ingredients including microbial enzymes continuously comes into contact with the skin starting in infancy. An impaired skin barrier has been suggested as a route of allergic sensitization. We hypothesized that exposure of skin to LD damages the skin barrier resulting in systemic sensitization to allergens that enter through the impaired skin barrier. Mouse skin samples exposed in vitro to microbial proteases or LD exhibited physical damage, which was more pronounced in neonatal skin as compared to adult skin. Exposure of the skin to microbial proteases in vitro resulted in an increase in the levels of interleukin (IL)-33 and thymic stromal lymphopoietin (TSLP). BALB/c wild type mice epicutaneously exposed to LD and ovalbumin (OVA) showed an increase in levels of transepidermal water loss, serum OVA-specific immunoglobulin (Ig) G1 and IgE antibodies, and a local increase of Il33, Tslp, Il4 and Il13 compared with LD or OVA alone. Following intranasal challenge with OVA, mice epicutaneously exposed to LD showed an increase in allergen-induced esophageal eosinophilia compared with LD or OVA alone. Collectively, these results suggest that LD may be an important factor that impairs the skin barrier and leads to allergen sensitization in early life, and therefore may have a role in the increase in allergic disease.

Topics: Allergens; Animals; Dermatitis, Atopic; Detergents; Eosinophilia; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Peptide Hydrolases

In our clinical work, some patients with type I hypersensitivity could be detected protein in their urine. This study focused on the early renal injury in patients with type I hypersensitivity.. From 43 type I hypersensitivity patients with proteinuria, 10 patients were randomly selected for mass spectrometry analysis of 24-h urine together with 5 healthy volunteers. Mice were vaccinated with Dermatophagoides farina (Der f) and ovalbumin (OVA) were used as antigen to establish the type I hypersensitivity animal models.. The urine protein of hypersensitivity patients was significantly increased in the alpha-1-microglobulin/ bikunin precursor (Protein AMBP) (t = 3.140, P = 0.008), retinol binding protein 4 (RBP4) (t = 2.426, P = 0.031), kininogen-1 (t = 2.501, P = 0.027), and transferrin appeared only in patients' urine. After immunizing mice with antigens, significant increases of the total serum immunoglobulin E (IgE) were observed in both Der f (86.92 ± 36.01 U/mL, t = 5.231, P = 0.0004) and OVA group (34.65 ± 24.72 U/mL, t = 2.891, P = 0.0161) compared with the negative control group (2.68 ± 0.47 U/mL). Meanwhile, definite eosinophil infiltration around the impaired renal tubules as well as the bronchus in Der f mice were observed, and urine protein appeared. After stopping the allergen stimulation, proteinuria disappeared. Instead, when the mice were treated with the antigen again, proteinuria reappeared.. Our findings suggest that renal tubular damage in patients with type I hypersensitivity is reversible, and proteinuria disappears with allergy symptoms remission.

Topics: Allergens; Animals; Humans; Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Kidney; Mice; Ovalbumin; Proteinuria; Retinol-Binding Proteins, Plasma

When the drug induces the organism to produce a type Ⅰ allergic reaction, the combination of IgE and mast cells results in the degranulation of the mast cells. Release of vasoactive substances, increase in vascular permeability, and exudation of intravascular substances outside the blood vessels. Based on this pathophysiological mechanism, a mouse model that can objectively and quantitatively assess the allergic response to the injection has been established. ICR mice were sensitised by intraperitoneal injection of different doses of OVA once every two days for three times. 14 days after the last sensitization, a combination OVA solution of 4 times the sensitizing dose and Evans blue were injected intravenously into mice for the challenge. Compared with the normal group, OVA 0.625/2.5, 1.25/5, 2.5/10, 5/20 mg·kg~(-1) sensitized and challenged can induce allergic reactions mainly manifested by blue staining of the auricle in mice. Direct injection of OVA intravenously did not cause an auricular blue colouration reaction in mice. The passive cutaneous anaphylaxis reaction in mice was conducted with the aforementioned OVA-sensitized mouse serum, and there were obvious blue spots on the mouse's back. In addition, the content of anti-OVA-IgE in 5 mg·kg~(-1) OVA-sensitized mice was significantly increased. Ears and lungs of mice sensitized to OVA showed evident exudation inflammation. Significantly elevated inflammatory factors(VEGF and IL-10) were also detected in the serum of OVA-sensitized mice. The equivalent dose of OVA caused obvious allergic reactions in both guinea pigs and mice. Compared with nude mice, ICR and BALB/c mice are more sensitive to OVA sensitization. Injections of selected TCMI did not induce type Ⅰ allergic reactions in mice and guinea pigs, but there was a risk of inducing pseu-doallergic reactions in mice. The model is problematic and may well reflect the sensitization effect of allergens. It obtains the benefits of simple operation, accuracy, low cost, easy extension, and high repeatability. It is suitable for predicting and researching for IgE-dependent type Ⅰ allergic reactions.

Topics: Allergens; Animals; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Mice, Nude; Ovalbumin

Ocular allergy leads to acute and chronic inflammation that may deteriorate the conjunctiva and other ocular tissue. The conjunctiva is covered with abundant lymphatic vessels but how the conjunctival lymphatic system patriciates in the development of allergic eye disease (AED) remains to be elucidated.. By using ovalbumin (OVA)+pertussis toxin (PTX) as a sensitizer followed by daily OVA challenges, we induced optimized AED manifestations in mice. We show that conjunctival lymphatics underwent significant expansion after 28 days of chronic OVA challenge, and this process can be prevented by inducible genetic ablation of lymphatic Vegfr3. Through transcriptomic profile analysis in combination with histopathological examinations, we found that pro-lymphangiogenic VEGFR3 signal promoted allergy-induced activation of T helper 2 (Th2) type immune responses, including antigen presentation, and Th2 cells, B cells and mast cell-related pathways in the conjunctiva, thereby critically contributing to the immunoglobulin E (IgE) production and AED manifestations. As a result, ocular allergy can be alleviated by genetic inhibition of lymphatic Vegfr3. Interestingly, pro-lymphangiogenic VEGFR3 signal did not appear to affect the obstruction of meibomian glands (MGs) or the activation of Th17 type and neutrophil pathways that are associated with AED.. These data reveal the key role of pro-lymphangiogenic VEGFR3 signaling in the development of AED and provide experimental evidence that VEGFR3 inhibition may be useful in treating ocular allergy in patients.

Topics: Animals; Disease Models, Animal; Eye Diseases; Hypersensitivity; Lymphangiogenesis; Mice; Ovalbumin; Vascular Endothelial Growth Factor Receptor-3

Anemoside B4 (AB4) is a natural triterpenoid abundant in the roots of Pulsatilla chinensis. Although various biological activities have been widely attributed to AB4, few studies have focused on its antiallergic effects. In this study the inhibitory effects of AB4 on immunoglobulin E (IgE)-mediated allergic responses were investigated, both in vitro and in vivo, and the mechanism of its effects. IgE-mediated passive cutaneous anaphylaxis was used to elucidate the antiallergic effects of AB4 in vivo. The degranulation assay, calcium imaging, and cytokine and chemokine release in the laboratory of allergic disease 2 (LAD2) cell line were used to evaluate the antiallergic effect of AB4 in vitro. Pathological staining was performed to analyze angiectasis. Western blot analysis was performed to investigate the downstream signaling pathways. AB4 dose-dependently attenuated ovalbumin/IgE-induced paw swelling in mice, and reduced the serum concentrations of tumor necrosis factor-alpha and C-C motif chemokine 2. In addition, AB4 suppressed IgE-mediated LAD2 cell degranulation, calcium influx, and PLC/IP3 and JAK/STAT3 phosphorylation. Our results suggest that AB4 inhibits allergic reactions through the PLC/IP3 and JAK/STAT3 pathways.

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Calcium; Cell Degranulation; Congenital Disorders of Glycosylation; Cytokines; Hypersensitivity; Immunoglobulin E; Mast Cells; Mice; Ovalbumin; Passive Cutaneous Anaphylaxis; Saponins; Triterpenes; Tumor Necrosis Factor-alpha

Innovations in oral immunotherapy have greatly advanced the therapeutic control of allergies. However, these therapeutic effects suffer from the fact that the amount of antigen delivered to antigen-presenting cells is limited given the formulations that are currently available. We recently designed a cell-penetrating albumin and found that this modified albumin enters cells via the induction of macropinocytosis. Herein, we report on a novel system for delivering antigens based on cell-penetrating albumin-inducible macropinocytosis that allows larger amounts of antigens to be delivered to antigen-presenting cells. A treatment with cell-penetrating albumin significantly increased the permeability of ovalbumin (45 kDa) or dextran (2000 kDa) on monolayers derived from human oral squamous carcinoma cells. Flow cytometric analyses showed that the cell-penetrating albumin treatment resulted in a significant elevation in the amount of dextran that was delivered to two types of antigen-presenting cells. Finally, mice that had been sensitized by Japanese cedar pollen extract (JCPE) and cell-penetrating albumin showed a decline in the frequency of nose-rubbing against a subsequent intranasal administration of JCPE. These findings suggest that the sublingual administration of cell-penetrating albumin efficiently delivers antigens to antigen-presenting cells via the induction of macropinocytosis, resulting in an enhancement in the therapeutic effect of sublingual immunotherapy.

Topics: Administration, Sublingual; Allergens; Animals; Antigens; Dextrans; Humans; Hypersensitivity; Mice; Ovalbumin

Infants with respiratory syncytial virus (RSV)-associated bronchiolitis are at increased risk of childhood asthma. Recent studies demonstrated that certain infections induce innate immune memory (also termed trained immunity), especially in macrophages, to respond more strongly to future stimuli with broad specificity, involving in human inflammatory diseases. Metabolic reprogramming increases the capacity of the innate immune cells to respond to a secondary stimulation, is a crucial step for the induction of trained immunity. We hypothesize that specific metabolic reprogramming of lung trained macrophages induced by neonatal respiratory infection is crucial for childhood allergic asthma.. To address the role of metabolic reprogramming in lung trained macrophages induced by respiratory virus infection in allergic asthma.. Neonatal mice were infected and sensitized by the natural rodent pathogen Pneumonia virus of mice (PVM), a mouse equivalent strain of human RSV, combined with ovalbumin (OVA). Lung CD11b. Proline metabolism reprogramming of trained macrophages induced by early respiratory infection combined with allergen sensitization contributes to development of allergic asthma in childhood. Proline metabolism could be a well target for prevention of allergic asthma in childhood.

Topics: Allergens; Animals; Asthma; Humans; Hypersensitivity; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Proline; Respiratory Syncytial Virus Infections; Respiratory Tract Infections

A new intervention was investigated for the induction of oral tolerance (OT) of OVA using narirutin by

Topics: Animals; Cytokines; Disease Models, Animal; Hypersensitivity; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; Receptors, CCR7; STAT5 Transcription Factor

Sulforaphane has been investigated in human pathologies and preclinical models of airway diseases. To provide further mechanistic insights, we explored L-sulforaphane (LSF) in the ovalbumin (OVA)-induced chronic allergic airways murine model, with key hallmarks of asthma. Histological analysis indicated that LSF prevented or reversed OVA-induced epithelial thickening, collagen deposition, goblet cell metaplasia, and inflammation. Well-known antioxidant and anti-inflammatory mechanisms contribute to the beneficial effects of LSF. Fourier transform infrared microspectroscopy revealed altered composition of macromolecules, following OVA sensitization, which were restored by LSF. RNA sequencing in human peripheral blood mononuclear cells highlighted the anti-inflammatory signature of LSF. Findings indicated that LSF may alter gene expression via an epigenetic mechanism which involves regulation of protein acetylation status. LSF resulted in histone and α-tubulin hyperacetylation in vivo, and cellular and enzymatic assays indicated decreased expression and modest histone deacetylase (HDAC) inhibition activity, in comparison with the well-known pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA). Molecular modeling confirmed interaction of LSF and LSF metabolites with the catalytic domain of metal-dependent HDAC enzymes. More generally, this study confirmed known mechanisms and identified potential epigenetic pathways accounting for the protective effects and provide support for the potential clinical utility of LSF in allergic airways disease.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Epigenesis, Genetic; Humans; Hypersensitivity; Leukocytes, Mononuclear; Mice; Ovalbumin

Chronic inflammation in the airway passage leads to the clinical syndrome of pediatric asthma. Allergic reactions caused by bacterial, viral, and fungal infection lead to the immune dis-balance which primes T helper cells (Th2), a specific cluster of differentiation 4 (CD4) T cell differentiation. This favors the Th2-specific response by activating the inter-leukin 4/interleukin 13 (IL-4/IL-13) cytokine signaling and further activates the secretion of immunoglobulin E (IgE). IL-13 develops bronchial asthma by elevating bronchial hyperresponsiveness and enables production of immunoglobulin M (IgM) and IgE. The present study aims to target IL-13 signaling using molecular docking and understanding molecular dynamic simulation (MDS) to propose a compelling candidate to treat asthma. We developed a library of available allergic drugs (n=20) and checked the binding affinity against IL-13 protein (3BPN.pdb) through molecular docking and confirmed the best pose binding energy of -3.84 and -3.71 for epinephrine and guaifenesin, respectively. Studying the interaction of hydrogen bonds and Van der Walls, it is estimated that electrostatic energy is sufficient to interact with the active site of the IL-13 and has shown to inhibit inflammatory signaling. These computational results confirm epinephrine and guaifenesin as potential ligands showing potential inhibitory activity for IL-13 signaling. This study also suggests the designing of a new ligand and screening of a large cohort of drugs, in the future, to predict the exact mechanism to control the critical feature of asthma.

Topics: Animals; Asthma; Child; Cytokines; Disease Models, Animal; Epinephrine; Guaifenesin; Humans; Hypersensitivity; Immunoglobulin E; Interleukin-13; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; Th2 Cells

Severe neutrophilic asthma is often characterized by persistent airway inflammation and irreversible airway remodeling, which are overstimulated by the high-mobility group box protein 1 (HMGB1). Although wogonin, an O-methylated flavone, has been widely used to treat inflammatory and allergic diseases, its therapeutic effects and potential mechanisms on severe neutrophilic asthma remain elusive.. To evaluate whether wogonin alleviates airway neutrophilia through inducing neutrophil apoptosis and attenuates airway smooth muscle cells (ASMCs) proliferation and migration.. The effect of wogonin on reducing neutrophilic airway inflammation, including neutrophil infiltration and inflammatory mediators, was examined in a mouse model of severe neutrophilic asthma sensitized with ovalbumin and lipopolysaccharide. Also, the effect of wogonin on inducing human neutrophil apoptosis was manifested using cellular morphology, flow cytometry, and caspase inhibition assays. Furthermore, the effect of wogonin on inhibiting HMGB1-mediated ASMCs proliferation and migration was determined.. Wogonin reduced the frequency of neutrophils and inhibited the production of multiple inflammatory mediators, including ovalbumin-specific IgE, tumor necrosis factor-α, interleukin-6, and HMGB1, in bronchoalveolar lavage fluid and lung tissues of the neutrophilic asthmatic mouse model. These data strongly support a significantly suppressed neutrophilic airway inflammation, functionally consistent to the relieved airway hyperresponsiveness by wogonin in vivo. Wogonin induced human neutrophil apoptosis in a dose-dependent manner by activating caspase-8 and caspase-3 in vitro. Wogonin pretreatment abolished HMGB1-induced ASMCs proliferation and migration, which can be explained by the inhibition of phosphorylation in the mitogen-activated protein kinase (MAPK) /Akt singling pathways.. Our findings demonstrate that wogonin augments caspase-dependent apoptosis in neutrophils to alleviate neutrophilic inflammatory responses and regulates intracellular signaling to inhibit HMGB1-mediated ASMCs activation, providing a promising therapeutic agent for severe neutrophilic asthma.

Topics: Animals; Apoptosis; Asthma; Cell Proliferation; HMGB1 Protein; Humans; Hypersensitivity; Inflammation; Inflammation Mediators; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Muscle, Smooth; Ovalbumin; Proto-Oncogene Proteins c-akt

The circadian clock is closely associated with inflammatory reactions. Increased inflammatory cytokine levels have been detected in the airways of nocturnal asthma. However, the mechanisms that contribute to the nocturnal increase in inflammatory responses and the relationship with circadian clock remain unknown.. Inflammatory cytokine levels were measured in asthma patients with and without nocturnal symptoms. Allergic airway disease was induced in mice by ovalbumin (OVA), and different periods of light/dark cycles were used to induce circadian rhythm disorders. Serum shock was used to stimulate the rhythmic expression in human bronchial epidermal cells (16HBE). The expression and oscillation of circadian clock genes and inflammatory cytokines in 16HBE cells subjected to brain and muscle ARNT-like protein-1 (BMAL1) and Forkhead Box A2 (FOXA2) knockdown and treatment with a FOXA2 overexpression plasmid were assessed.. Serum IL-6 was found to be significantly higher in asthmatic patients with nocturnal symptoms than those without nocturnal symptoms. The OVA-induced asthma model with a circadian rhythm disorder and 16HBE cells treated with serum shock showed an increase in IL-6 levels and a negative correlation with BMAL1 and FOXA2. The knockdown of BMAL1 resulted in a lower correlation between IL-6 and other rhythm clock genes. Furthermore, knockdown of the BMAL1 and FOXA2 in 16HBE cells reduced the expression and rhythmic fluctuations of IL-6.. Our findings suggest that there are increased IL-6 levels in nocturnal asthma resulting from inhibition of the BMAL1/FOXA2 signalling pathway in airway epithelial cells.

Topics: Animals; Asthma; Cytokines; Hepatocyte Nuclear Factor 3-beta; Humans; Hypersensitivity; Interleukin-6; Mice; Ovalbumin

Global air pollution and diesel exhaust particles (DEPs) generated by intratracheal instillation aggravate asthma. In this study, we evaluated the effect of probiotics via tracheal- or oral-route administration on allergies or asthma. We continuously perfused rats daily, using the oral and tracheal routes, with approximately 10

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Rats

Titanium dioxide nanoparticles (TiO

Topics: Animals; Apoptosis; Asthma; bcl-2-Associated X Protein; Bronchoalveolar Lavage Fluid; Carrier Proteins; Caspase 3; Cell Count; Cell Line; Chemical Phenomena; Cytokines; Disease Models, Animal; Gene Expression Regulation; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Inflammation Mediators; Lung; MAP Kinase Kinase Kinase 5; Mice; Mucus; Nanoparticles; Ovalbumin; Respiratory Hypersensitivity; RNA, Messenger; Thioredoxins; Titanium; Up-Regulation

Allergic asthma is well known as a common respiratory disorder comprising an allergic inflammatory nature and excessive immune characteristic.

Topics: Adenosine; Allergens; Animals; Asthma; Disease Models, Animal; DNA Methylation; Epigenesis, Genetic; Epigenome; Female; Gene Expression Profiling; Humans; Hypersensitivity; Immunity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction

CpG-oligodeoxynucleotides (CpG-ODNs) constitute an attractive alternative for asthma treatment. However, very little evidence is available from studies on the oral administration of CpG-ODNs in animals. Previously, we developed acid-resistant particles (named ODNcap) as an oral delivery device for ODNs. Here, we showed that free feeding of an ODNcap-containing feed prophylactically attenuates allergic airway inflammation, hyperresponsiveness, and goblet cell hyperplasia in an ovalbumin-induced asthma model. Using transcriptomics-driven approaches, we demonstrated that injury of pulmonary vein cardiomyocytes accompanies allergen inhalation challenge, but is inhibited by ODNcap feeding. We also showed the participation of an airway antimicrobial peptide (Reg3γ) and fecal microbiota in the ODNcap-mediated effects. Collectively, our findings suggest that daily oral ingestion of ODNcap may provide preventive effects on allergic bronchopulmonary insults

Topics: Administration, Oral; Animals; Antimicrobial Peptides; Bronchial Hyperreactivity; Female; Gastrointestinal Microbiome; Hypersensitivity; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; Pancreatitis-Associated Proteins; Pneumonia

Fine-tuning of immune receptor signaling is critical for the development and functioning of immune cells. Moreover, GM-CSF receptor (GM-CSFR) signaling plays an essential role in the development of certain myeloid lineage cells, including alveolar macrophages (AMs). However, the significance of fine-tuning of GM-CSFR signaling in AMs and its relevance in allergic inflammation have not been reported.. Our aim was to explore whether phosphatase Ssu72, originally identified as a regulator of RNA polymerase II activity, regulates AM development and allergic airway inflammation by regulating GM-CSF signaling.. To address these issues, we generated LysM-CreSsu72. Following GM-CSF stimulation, Ssu72 directly bound to the GM-CSFR β-chain in AMs, preventing phosphorylation. Consistently, mature Ssu72-deficient AMs showed higher phosphorylation of the GM-CSFR β-chain and downstream molecules, which resulted in greater dysregulation of cell cycle, cell death, cell turnover, mitochondria-related metabolism, and LPS responsiveness in AMs than in mature wild-type AMs. The dysregulation was restored by using a Janus kinase 2 inhibitor, which reduced GM-CSFR β-chain phosphorylation. LysM-CreSsu72. Our results demonstrate that Ssu72 fine-tunes GM-CSFR signaling by both binding to and reducing phosphorylation of GM-CSFR β-chain, thereby regulating the development, maturation, and mitochondrial functions of AMs and allergic airway inflammation.

Topics: Animals; Antigens, Dermatophagoides; CD11c Antigen; Cell Differentiation; Cells, Cultured; Disease Models, Animal; Humans; Hypersensitivity; Macrophages, Alveolar; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Phosphoprotein Phosphatases; Pyroglyphidae; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; Respiratory Hypersensitivity; Signal Transduction

Allergic asthma is a chronic inflammatory lung disease characterized by a Th2-type immune response pattern. The development of nonspecific immunotherapy is one of the primary goals for the control of this disease.. In this study, we evaluated the therapeutic effects of Lactococcus lactis-producing mycobacterial heat shock protein 65 (LLHsp65) in an ovalbumin (OVA)-induced allergic asthma model. OVA-challenged BALB/c mice were orally administrated with LLHsp65 for 10 consecutive days. The results demonstrate that LLhsp65 attenuates critical features of allergic inflammation, like airway hyperresponsiveness and mucus production. Likewise, the treatment decreases the pulmonary eosinophilia and the serum level of OVA-specific IgE. In addition to deviating immune responses towards Th1-cytokine profile, increase regulatory T cells, and cytokine levels, such as IL-6 and IL-10.. Our results reveal that the mucosal immunotherapy of LLHsp65 significantly reduces the overall burden of airway allergic inflammation, suggesting a promising therapeutic strategy for allergic asthma treatment.. This research reveals new perspectives on nonspecific immunotherapy based on the delivery of recombinant proteins by lactic acid bacteria to treat of allergic disorders.

Topics: Administration, Oral; Animals; Asthma; Bacterial Proteins; Bronchoalveolar Lavage Fluid; Chaperonin 60; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Inflammation; Lactococcus lactis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

Bronchial asthma (BA) is a chronic disease of the airways. The great majority of BA exacerbations are associated with respiratory viral infections. Recent findings point out a possible role of proinflammatory cytokine interleukin-33 (IL-33) in the development of atopic diseases. Although, little is known about the role of IL-33 in virus-induced BA exacerbations.. We used mouse models of RSV (respiratory syncytial virus)-induced inflammation exacerbation in OVA-sensitized mice and RSV infection alone in adult animals to characterize expression of il33 in the mouse lungs. Moreover, we studied the influence of il33 knockdown with intranasally administrated siRNA on the development of RSV-induced inflammation exacerbation. In addition, we evaluated the expression of IL33 in the ex vivo stimulated PBMCs from allergic asthma patients and healthy subjects with and without confirmed acute respiratory viral infection.. Using mouse models, we found that infection with RSV drives enhanced il33 mRNA expression in the mouse lung. Treatment with anti-il33 siRNA diminishes airway inflammation in the lungs (we found a decrease in the number of inflammatory cells in the lungs and in the severity of histopathological alterations) of mice with RSV-induced inflammation exacerbation, but do not influence viral load. Elevated level of the IL33 mRNA was detected in ex vivo stimulated blood lymphocytes of allergic asthmatics infected with respiratory viruses. RSV and rhinovirus were the most detected viruses in volunteers with symptoms of respiratory infection.. The present study provides additional evidence of the crucial role of the IL-33 in pathogenesis of RSV infection and virus-induced allergic bronchial asthma exacerbations.

Topics: Adolescent; Adult; Aged; Animals; Asthma; Disease Models, Animal; Female; Humans; Hypersensitivity; Inflammation; Interleukin-33; Leukocytes, Mononuclear; Lung; Male; Mice; Mice, Inbred BALB C; Middle Aged; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human; Respiratory Syncytial Viruses; RNA, Messenger; RNA, Small Interfering; Up-Regulation; Young Adult

Polymorphonuclear neutrophils (PMN) are one fraction of the major inflammatory cells in allergic asthma (asthma, in short); the role of PMN in the asthma pathogenesis is not fully understood yet. This study aims to investigate the effects of specific Ag-guiding exosomes on suppressing the neutrophil-dominant airway inflammation. In this study, BALB/c mice were immunized with ovalbumin plus complete Freund adjuvant to induce an asthma model featured with neutrophil-dominant lung inflammation. The Ag specific PMN (sPMN)-targeting exosomes (tExo), that were exosomes carrying a complex of specific Ag/anti-CD64 Ab and Fas ligand, were constructed to be used to alleviate neutrophilic asthma in mice. We found that sPMNs were the major cellular component in bronchoalveolar lavage fluid (BALF) in asthma mice, while less than 3% PMNs in naive control mice. The sPMNs expressed higher levels of CD64, which formed complexes with Ag-specific IgG (sIgG). The sIgG/CD64 complex-carrying PMNs could be activated upon exposing to specific Ags. Exposure to tExos induced Ag-specific PMNs apoptosis. Administration of tExos efficiently suppressed experimental asthma. We conclude that a fraction of sPMN was identified in the airway of asthma mice. The sPMNs could be activated upon exposure to specific Ags. tExos could induce sPMNs apoptosis, that show the translational potential in the treatment of asthma.

Topics: Animals; Antibodies; Antigens; Apoptosis; Asthma; Bronchoalveolar Lavage Fluid; Drug Carriers; Exosomes; Freund's Adjuvant; Hypersensitivity; Immunoglobulin G; Lung; Mice; Mice, Inbred BALB C; Nanoparticles; Neutrophils; Ovalbumin; Pneumonia; Receptors, IgG; Vaccines

Growing evidence shows that enhancer of zeste homolog 2 (EZH2) plays a role in various physiological functions and cancer pathogenesis. However, its contribution to allergic diseases remains controversial. We sought to investigate the role of EZH2 in the pathogenesis of allergic airway inflammation.. 3-Deazaneplanocin A (DZNep), an indirect inhibitor of EZH2, was administered via intraperitoneal injection in an ovalbumin (OVA)-induced murine model of allergic airway inflammation. The expression of EZH2 in the allergic airway tissues was examined by immunohistochemistry (IHC) and western blot. The inflammatory cell infiltration and the goblet cell hyperplasia in the murine nose and lung were detected by hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining. Levels of cytokines, including IL-4, IFN-γ, IL-6, and IL-10, were evaluated in the bronchoalveolar lavage fluid (BALF) using Enzyme-linked immune sorbent assay (ELISA).. EZH2 expression was inhibited by DZNep treatment (P < 0.05). The administration of DZNep significantly inhibited the inflammatory cell infiltration (P < 0.0001) and goblet cell hyperplasia (P < 0.001). Moreover, it suppressed the secretion of IL-4 (P < 0.0001) and IL-6 (P < 0.01) in the BALF.. Our findings demonstrate that DZNep attenuates allergic airway inflammation and could be a new therapeutic option for allergic rhinitis and asthma.

Topics: Adenosine; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Traditionally, the roots of Angelica reflexa B.Y.Lee (AR) have been used to treat cough, phlegm, neuralgia, and arthralgia in Northeast Asia.. The anti-asthmatic effect of AR root extract (ARE) was determined using a murine airway allergic inflammation model and the primary T cell polarization assay.. To evaluate the anti-asthmatic effect of ARE, inflammatory cell infiltration was determined histologically and inflammatory mediators were measured in bronchoalveolar lavage fluid (BALF). Furthermore, the effects of AREs on Th2 cell differentiation and activation were determined by western blotting and flow cytometry.. Asthmatic phenotypes were alleviated by ARE treatment, which reduced mucus production, inflammatory cell infiltration (especially eosinophilia), and type 2 cytokine levels in BALF. ARE administration to mice reduced the number of activated Th2 (CD4. Our findings indicate that the anti-asthmatic effect of AREs is mediated by the reduction in Th2 cell activation by regulating IRF4.

Topics: Angelica; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; GATA3 Transcription Factor; Hypersensitivity; Interferon Regulatory Factors; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; Plant Roots; Pneumonia; Pulmonary Eosinophilia; RAW 264.7 Cells; Th2 Cells

Lung nociceptor neurons amplify immune cell activity and mucus metaplasia in response to an inhaled allergen challenge in sensitized mice.. We sought to identify the cellular mechanisms by which these sensory neurons are activated subsequent to allergen exposure.. We used calcium microscopy and electrophysiologic recording to assess whether vagal neurons directly respond to the model allergen ovalbumin (OVA). Next, we generated the first nociceptor-specific FcεR1γ knockdown (TRPV1. Lung-innervating jugular nodose complex ganglion neurons express the high-affinity IgE receptor FcεR1, the levels of which increase in OVA-sensitized mice. FcεR1γ-expressing vagal nociceptor neurons respond directly to OVA complexed with IgE with depolarization, action potential firing, calcium influx, and neuropeptide release. Activation of vagal neurons by IgE-allergen immune complexes, through the release of substance P from their peripheral terminals, directly amplifies T. Allergen sensitization triggers a feedforward inflammatory loop between IgE-producing plasma cells, FcεR1-expressing vagal sensory neurons, and T

Topics: Allergens; Animals; Calcium; Disease Models, Animal; Disease Susceptibility; Gene Expression; Genetic Predisposition to Disease; Hypersensitivity; Mice; Mice, Knockout; Neurons; Nociceptors; Ovalbumin; Receptors, IgE; Respiratory Mucosa; Substance P; Vagus Nerve

RGFP966 is a selective inhibitor of histone deacetylase 3 (HDAC3) playing crucial roles in triggering allergic and inflammatory responses. Whereas, its role in allergic rhinitis (AR) remains uncertain. This study sought to illustrate the role and mechanism of HDAC3 inhibitor RGFP966 on allergic and inflammatory responses in murine AR. RGFP966 administration was applied on murine AR. HE staining, PAS staining, toluidine blue staining, immunohistochemistry staining and real-time PCR methods were used to assess eosinophils, goblet cells, mast cells, HDAC3 positive cells and mRNA levels in nasal tissues of mice. HDAC3 activities in nasal tissues were quantified with HDAC3 Activity Assay Kit. We collected blood and nasal lavage fluid (NLF) of mice for assaying IgE, inflammatory cytokines and inflammatory cells. Results indicated that RGFP966 intervention attenuated sneezing, nose rubbing, IgE, inflammatory cytokines, eosinophils, goblet cells, mast cells, inflammatory cells, HDAC3 levles and activities in RGFP966 treated mice. In conclusion, RGFP966 might reduce HDAC3 expression and HDAC3 activities, and then eosinophils and mast cells recruitment, goblet cells proliferation and inflammatory cytokines levels are decreased, resulting in the alleviation of allergic and inflammatory responses in AR mice.

Topics: Acrylamides; Allergens; Animals; Anti-Inflammatory Agents; Disease Models, Animal; Female; Histone Deacetylases; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Mice; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Phenylenediamines

Topics: Allergens; Animals; Antigens, CD; CD83 Antigen; Cell Differentiation; Disease Models, Animal; Epithelial Cells; Exosomes; Hypersensitivity; Immune Tolerance; Immunoglobulins; Lymphocyte Activation; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Respiratory Hypersensitivity; Respiratory Mucosa; T-Lymphocytes, Regulatory

Allergic airway inflammation is heterogeneous with variability in immune phenotypes observed across asthmatic patients. Inflammation has been thought to directly contribute to airway remodeling in asthma, but clinical data suggest that neutralizing type 2 cytokines does not necessarily alter disease pathogenesis. Here, we utilized C57BL/6 and BALB/c mice to investigate the development of allergic airway inflammation and remodeling. Exposure to an allergen cocktail for up to 8 weeks led to type 2 and type 17 inflammation, characterized by airway eosinophilia and neutrophilia and increased expression of chitinase-like proteins in both C57BL/6 and BALB/c mice. However, BALB/c mice developed much greater inflammatory responses than C57BL/6 mice, effects possibly explained by a failure to induce pathways that regulate and maintain T-cell activation in C57BL/6 mice, as shown by whole lung RNA transcript analysis. Allergen administration resulted in a similar degree of airway remodeling between mouse strains but with differences in collagen subtype composition. Increased collagen III was observed around the airways of C57BL/6 but not BALB/c mice while allergen-induced loss of basement membrane collagen IV was only observed in BALB/c mice. This study highlights a model of type 2/type 17 airway inflammation in mice whereby development of airway remodeling can occur in both BALB/c and C57BL/6 mice despite differences in immune response dynamics between strains. Importantly, compositional changes in the extracellular matrix between genetic strains of mice may help us better understand the relationships between lung function, remodeling and airway inflammation.

Topics: Airway Remodeling; Allergens; Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin

Eosinophils are terminally differentiated cells derived from hematopoietic stem cells (HSCs) in the bone marrow. Several studies have confirmed the effective roles of eosinophils in asthmatic airway pathogenesis. However, their regulatory functions have not been well elucidated. Here, increased C-C chemokine ligand 6 (CCL6) in asthmatic mice and the human orthologs CCL15 and CCL23 that are highly expressed in asthma patients are described, which are mainly derived from eosinophils. Using Ccl6 knockout mice, further studies revealed CCL6-dependent allergic airway inflammation and committed eosinophilia in the bone marrow following ovalbumin (OVA) challenge and identified a CCL6-CCR1 regulatory axis in hematopoietic stem cells (HSCs). Eosinophil differentiation and airway inflammation were remarkably decreased by the specific CCR1 antagonist BX471. Thus, the study identifies that the CCL6-CCR1 axis is involved in the crosstalk between eosinophils and HSCs during the development of allergic airway inflammation, which also reveals a potential therapeutic strategy for targeting G protein-coupled receptors (GPCRs) for future clinical treatment of asthma.

Topics: Adolescent; Adult; Aged; Animals; Asthma; Bone Marrow; Cell Differentiation; Chemokines, CC; Eosinophils; Female; Healthy Volunteers; Hematopoietic Stem Cells; Humans; Hypersensitivity; Inflammation; Lung; Macrophage Inflammatory Proteins; Male; Mice; Mice, Knockout; Middle Aged; Ovalbumin; Phenylurea Compounds; Piperidines; Receptors, CCR1; Signal Transduction; Young Adult

Asthma is a common respiratory disease currently affecting more than 300 million worldwide and is characterized by airway inflammation, hyperreactivity, and remodeling. It is a heterogeneous disease consisting of corticosteroid-sensitive T-helper cell type 2-driven eosinophilic and corticosteroid-resistant, T-helper cell type 17-driven neutrophilic phenotypes. One pathway recently described to regulate asthma pathogenesis is cholesterol trafficking. Scavenger receptors, in particular SR-BI (scavenger receptor class B type I), are known to direct cellular cholesterol uptake and efflux. We recently defined SR-BI functions in pulmonary host defense; however, the function of SR-BI in asthma pathogenesis is unknown. To elucidate the role of SR-BI in allergic asthma, SR-BI-sufficient (SR-BI

Topics: Adrenal Insufficiency; Animals; Asthma; CD36 Antigens; Hypersensitivity; Inflammation; Interleukin-17; Lung; Male; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Pyroglyphidae; Th17 Cells

Inhibition of allergic airway diseases (AAD) by immunomodulation of the adaptive immune system through restoration of the enteric dysbiosis is an emerging therapeutic strategy. Patients with allergic rhinitis (n = 6) and healthy controls (n = 6) were enrolled, and gut microbiome composition analysis was performed by 16S rDNA sequencing. We also established an ovalbumin (OVA)-induced allergic airway inflammation murine model. Dysbiosis of the gut flora was observed in both AAD patients and the mice, with the decrease of the biodiversity and the quantity of the Bacteroidetes phylum. Oral application of

Topics: Allergens; Animals; Bacteroides thetaiotaomicron; Cells, Cultured; Cytokines; Gastrointestinal Microbiome; Humans; Hypersensitivity; Immunomodulation; Inducible T-Cell Co-Stimulator Protein; Inflammation; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Respiratory Hypersensitivity; Respiratory System; T-Lymphocytes, Regulatory; Th2 Cells

Asthma is an allergic chronic respiratory disease that affects more than 300 million people around the world. Dysbiosis of intestinal commensal microbiota influences the development of asthma. Dectin-1 (gene symbol:

Topics: Animals; Bacteria; Bronchoalveolar Lavage Fluid; Gastrointestinal Microbiome; Hypersensitivity; Intestines; Lactobacillus; Lectins, C-Type; Lung; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Pneumonia; RNA, Ribosomal, 16S; T-Lymphocytes, Regulatory

Microglia play key roles in synaptic pruning, which primarily occurs from the postnatal period to adolescence. Synaptic pruning is essential for normal brain development and its impairment is implicated in neuropsychiatric developmental diseases such as autism spectrum disorders (ASD). Recent epidemiological surveys reported a strong link between ASD and atopic/allergic diseases. However, few studies have experimentally investigated the relationship between allergy and ASD-like manifestations, particularly in the early postnatal period, when allergic disorders occur frequently. Therefore, we aimed to characterize how allergic inflammation in the early postnatal period influences microglia and behavior using mouse models of short- and long-term airway allergy. Male mice were immunized by an intraperitoneal injection of aluminum hydroxide and ovalbumin (OVA) or phosphate-buffered saline (control) on postnatal days (P) 3, 7, and 11, followed by intranasal challenge with OVA or phosphate-buffered saline solution twice a week until P30 or P70. In the hippocampus, Iba-1-positive areas, the size of Iba-1-positive microglial cell bodies, and the ramification index of microglia by Sholl analysis were significantly smaller in the OVA group than in the control group on P30 and P70, although Iba-1-positive microglia numbers did not differ significantly between the two groups. In Iba-1-positive cells, postsynaptic density protein 95 (PSD95)-occupied areas and CD68-occupied areas were significantly decreased on P30 and P70, respectively, in the OVA group compared with the control group. Immunoblotting using hippocampal tissues demonstrated that amounts of PSD95, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor 2, and N-methyl-D-aspartate (NMDA) receptor 2B were significantly increased in the OVA group compared with the control group on P70, and a similar increasing trend for PSD95 was observed on P30. Neurogenesis was not significantly different between the two groups on P30 or P70 by doublecortin immunohistochemistry. The social preference index was significantly lower in the three chamber test and the number of buried marbles was significantly higher in the OVA group than in the control group on P70 but not on P30, whereas locomotion and anxiety were not different between the two groups. Compared with the control group, serum basal corticosterone levels were significantly elevated and hippocampal glucocorticoid receptor (GR) amounts and nuclear GR

Topics: Animals; Autistic Disorder; Disease Models, Animal; Hypersensitivity; Inflammation; Male; Mice; Microglia; Ovalbumin

Asthma is a chronic inflammatory disease, characterized by airway inflammation, hyperresponsiveness, and bronchial smooth muscle contraction. Qingfei Xiaoyan Wan (QFXYW), a traditional Chinese formula, has been shown to exert anti-asthma effects and immune response in multiple diseases.. In this study, we evaluated the therapeutic mechanism of QFXYW in the suppression of allergic asthma by integrating of transcriptomics and system pharmacology.. BALB/c mice were sensitized with ovalbumin (OVA) to establish the allergic asthma model, and its success was confirmed with behavioral observations. Lung histopathological analysis, inflammatory pathology scores, transcription factors were used to evaluate the effects of QFXYW on allergic asthma. The therapeutic mechanism of QFXYW in treating allergic asthma through integrated transcriptomics and system pharmacology was then determined: hub genes were screened out by topological analysis and functional enrichment analysis were performed to identify key signaling pathway. Subsequently, quantitative RP-PCR and protein array were performed to detect the mRNA of hub genes and to predict the key pathway in OVA-induced allergic asthma, respectively.. Our results demonstrated that QFXYW could significantly attenuate inflammatory cell infiltration, mucus secretion, and epithelial damage. The transcriptomics analysis found the six hub genes with the highest values- CXCL10, CXCL2, CXCL1, IL-6, CCL-5, and CCL-4 were screened out. Functional enrichment analysis showed that the differentially expressed genes (DEGs) were mainly enriched in the inflammatory response and cytokine signaling pathway. Moreover, the quantitative RT-PCR verification experiment found the CXCL2 and CXCL1 were significantly suppressed after treatment with QFXYW. The results of protein array showed that QFXYW inhibited the multi-cytokines of OVA-induced allergic asthma via cytokine signaling pathway.. QFXYW may have mediated OVA-induced allergic asthma mainly through the hub genes CXCL2, CXCL1, and the cytokine signaling pathway. This finding will offer a novel strategy to explore effective and safe mechanism of Traditional Chinese Medicine (TCM) formula to treat allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Drugs, Chinese Herbal; Female; Gene Expression Regulation; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Transcriptome

Topics: Allergens; Animals; Anti-Allergic Agents; Asthma; Dendritic Cells; Disease Models, Animal; Gastrointestinal Microbiome; Humans; Hypersensitivity; Intestinal Mucosa; Lactobacillus; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; T-Lymphocytes, Regulatory

This study is to determine the role and mechanism of cryptotanshinone (CTS) in allergic airway inflammation. Asthma induced by OVA was established in BALB/c mice. We found increased airway hyperresponsiveness (AHR), increased inflammatory cell infiltration, elevated levels of TNF-α, interleukin-1β (IL-1β), IL-4, IL-5, IL-6 and IL-13, decreased interferon gamma (IFN-γ) in lung tissue, increased content of total immunoglobulin E (IgE), OVA specific IgE, Eotaxin, ICAM-1, VCAM-1, nuclear factor-kappaB (NF-κB) and phosphorylation of p38 MAPK in lung tissue. However, the administration of CTS significantly decreased AHR in asthmatic mice, reduced inflammation around the bronchioles and inflammatory cells around airway, regulated cytokine production, reduced the total IgE and OVA-specific IgE levels, and inhibited NF-κB activation and p38 MAPK phosphorylation. In vitro experiments in 16 HBE cells revealed that CTS attenuated CAM-1 and IL-6 expression. These results indicate that CTS alleviates allergic airway inflammation by modulating p38 MAPK phosphorylation and NF-κB activation.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Cytokines; Drugs, Chinese Herbal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation

1-nitropyrene (1-NP) is widespread in the environment, as a typical nitrated polycyclic aromatic hydrocarbon. The purpose of this research was to explore the effects of gestational 1-NP exposure on susceptibility of allergic asthma in offspring. Maternal mice were exposed to 1-NP (100 μg kg

Topics: Animals; Asthma; Disease Models, Animal; Environmental Pollutants; Female; Goblet Cells; Humans; Hypersensitivity; Lung; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Pyrenes

Allergic asthma is a chronic inflammatory airway disease driven predominantly by a T. We investigated potential therapeutic effects of selective inhibition of this pathway in mice with established allergic airway disease. We further investigated whether IL-4Rα disruption in systemically sensitized mice can prevent the onset of the disease.. Inducible deletion of IL-4Rα demonstrated therapeutic effects, on established allergic airway disease, and prevented the development of ovalbumin-induced airway hyperreactivity, eosinophilia, and goblet cell metaplasia in allergen-sensitized mice. Interestingly, IL-4Rα knockdown after allergic sensitization did not induce T. Abrogation of IL-4Rα signaling after allergic sensitization would have significant therapeutic benefit for T

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Scrophularia buergeriana Miq. (Scrophulariaceae) (SB) has been used as an oriental medicine for the treatment of inflammatory diseases, such as neuritis and pharyngolaryngitis.. We explored the therapeutic effects of S. buergeriana ethanol extract (SBE) on airway inflammation in ovalbumin (OVA)-induced asthmatic mice and lipopolysaccharide (LPS)-stimulated RAW264.7 cells.. Mice were intraperitoneally injected with OVA on days 0 and 14 to elevate the immune response. On days 21 to 23, the mice were challenged with OVA solution and SBE (20 and 40 mg/kg) was administered daily by oral gavage from days 18 to 23. RAW264.7 cells were pretreated with SBE 1 h before LPS stimulation.. SBE administration effectively suppressed inflammatory cell infiltration, the expression of interleukin (IL)-5, IL-13, and IL-17, immunoglobulin E, and airway hyperresponsiveness in an OVA-induced allergic asthma model. A reduction in histological alterations, including airway inflammation and mucus hypersecretion, was observed. These effects of SBE were accompanied by a decrease in matrix metalloproteinase-9 (MMP-9) expression and nuclear factor kappa B (NF-κB) phosphorylation. These responses were observed in LPS-stimulated RAW264.7 cells. SBE treatment reduced the mRNA expression of tumor necrosis factor (TNF)-α, IL-6, and MMP-9, and NF-κB phosphorylation, in LPS-stimulated RAW264.7 cells.. Our results indicated that SBE effectively attenuated airway inflammation in an OVA-induced allergic asthma model. These properties of SBE were thought to be involved in the suppression of NF-κB phosphorylation, suggesting that the material has the potential to regulate the development of allergic asthma.

Topics: Animals; Asthma; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lipopolysaccharides; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phosphorylation; Plant Extracts; RAW 264.7 Cells; Scrophularia

Translationally controlled tumor protein (TCTP), also called histamine releasing factor, is an evolutionarily conserved multifunctional protein in eukaryotes. We previously reported that extracellular TCTP acquires its cytokine-like function following dimerization. This study aims to identify the functional domain involved in the cytokine-like function of dimerized TCTP (dTCTP). We performed X-ray crystallographic studies and a deletion mutant of dTCTP which lacks the flexible loop domain. Synthetic peptides corresponding to TCTP domains and antibodies developed against them were examined for the anti-allergic effect. In an OVA-induced airway inflammation mouse model, inhibitory effect of synthetic peptides was evaluated. dTCTP was mediated by dimers between Cys172s of TCTP monomers. Synthetic peptides corresponding to the flexible loop and helix 2 domain of TCTP, and antibodies against them inhibited dTCTP-induced IL-8 release. In particular, the TCTP mutant lacking the flexible loop domain decreased the inflammatory cytokine activity of dTCTP. We conclude that the flexible loop and helix 2 domain of TCTP are the functional domains of dTCTP. They may have the potential to be therapeutic targets in the suppression of allergic reactions induced by dTCTP.

Topics: Animals; Anti-Allergic Agents; Biomarkers, Tumor; Crystallography, X-Ray; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Peptide Fragments; Protein Domains; Protein Multimerization; Signal Transduction; Tumor Protein, Translationally-Controlled 1

MiR-146a has been shown to negatively regulate innate immune, inflammatory response and antiviral pathway, however, its role in the tolerogenic responses remains largely unknown. This study aimed to investigate the role of miR-146a in the OVA-induced allergic inflammation of dendritic cells (DCs).. Bone marrow-derived DCs (BMDCs) were treated with OVA (100 µg/ml) for 24 h. MiR-146a expressions were assessed by quantitative RT-PCR. BMDCs were transfected with miR-146a mimics or inhibitor. Cell surface markers were analyzed by flow cytometry. Cytokine levels were determined by ELISA assay. Mixed lymphocyte culture assay was adopted to assess CD4 + T-cell differentiation. The 3' UTR luciferase reporter assay was utilized to determine the miRNA target sequence.. OVA treatment significantly up-regulated miR-146a in BMDCs in a dose- and time-dependent manner. In the OVA-treated DCs, overexpression of miR-146a (mimics transfection) down-regulated the surface markers (CD80, CD86) and increased production of anti-inflammatory cytokines TGF-β1 and IL-10 but decreased pro-inflammatory cytokine IL-12. MiR-146a overexpression promoted immature DC to induce regulatory T cells (Treg) differentiation. By contrast, transfection of miR-146a inhibitor into DC exhibited the opposite trends. Notch1 was a direct target of miR-146a, and Notch1 knock-down induced similar effects as miR-146a mimics transfection in BMDCs. Moreover, the effect of miR-146a inhibitor on OVA-induced DC was attenuated by Notch1 knock-down.. miRNA-146a promoted tolerogenic properties of DCs, at least partially, through targeting Notch1 signaling.

Topics: Allergens; Animals; Cell Differentiation; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Humans; Hypersensitivity; Immune Tolerance; Inflammation; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Receptor, Notch1; RNA, Small Interfering; Signal Transduction; T-Lymphocytes, Regulatory

The use of antibiotics during pregnancy is associated with increased allergic asthma risk in the offspring, and given that approximately 25% of pregnant women are prescribed antibiotics, it is important to understand the mechanisms contributing to this phenomenon. Currently, there are no studies that directly test this association experimentally. Our objective was to develop a mouse model in which antibiotic treatment during pregnancy results in increased offspring asthma susceptibility.. Pregnant mice were treated daily from gestation day 8-17 with an oral solution of the antibiotic vancomycin, and three concentrations were tested. At weaning, offspring were subjected to an adjuvant-free experimental asthma protocol using ovalbumin as an allergen. The composition of the gut microbiome was determined in mothers and offspring with samples collected from five different time points; short-chain fatty acids were also analyzed in allergic offspring.. We found that maternal antibiotic treatment during pregnancy was associated with increased offspring asthma severity in a dose-dependent manner. Furthermore, maternal vancomycin treatment during pregnancy caused marked changes in the gut microbiome composition in both mothers and pups at several different time points. The increased asthma severity and intestinal microbiome changes in pups were also associated with significantly decreased cecal short-chain fatty acid concentrations.. Consistent with the "Developmental Origins Hypothesis," our results confirm that exposure to antibiotics during pregnancy shapes the neonatal intestinal environment and increases offspring allergic lung inflammation.

Topics: Animals; Anti-Bacterial Agents; Asthma; Female; Humans; Hypersensitivity; Mice; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects

Atopic dermatitis skin lesions demonstrate increased expression of IL-25 by keratinocytes and increased numbers of type 2 innate lymphoid cells (ILC2s) that express high levels of IL-25 receptor (IL-25R). IL-13 is expressed in atopic dermatitis skin lesions and plays an important role in pathogenesis of the disease.. Our aim was to determine the role of IL-25 and ILC2s in a mouse model of antigen-driven allergic skin inflammation.. Wild-type mice; mice that express an Il13-driven enhanced green fluorescent protein; and mice that lack IL-25R, IL-25 in keratinocytes, or IL-13 or IL-25R in ILC2s were subjected to acute or chronic epicutaneous sensitization with ovalbumin. Sensitized skin was examined by histology for epidermal thickening. Cellular infiltrates were analyzed for surface markers and intracellular expression of enhanced green fluorescent protein by flow cytometry. Gene expression was quantitated by RT quantitative PCR.. In both acute and chronic antigen-driven allergic skin inflammation, signaling by keratinocyte-derived IL-25 in ILC2s is important for epidermal hyperplasia, dermal infiltration by CD4. ILC2 activation by IL-25 is essential for IL-13 expression at sites of allergic skin inflammation.

Topics: Allergens; Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Dermatitis, Atopic; Female; Gene Expression; Green Fluorescent Proteins; Hypersensitivity; Inflammation; Interleukin-13; Interleukins; Keratinocytes; Lymphocytes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Skin; Th2 Cells

Topics: Administration, Oral; Allergens; Animals; CD11c Antigen; Clostridiales; Dendritic Cells; Disease Models, Animal; Female; Gram-Positive Bacterial Infections; Humans; Hypersensitivity; Immunity, Cellular; Immunosuppression Therapy; Interleukin-2 Receptor alpha Subunit; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Airway remodeling happens in childhood asthma, in parallel with, but not necessarily subsequent to, airway inflammation. The differentiation of airway epithelial cells into myofibroblasts via epithelial-mesenchymal-transition (EMT) is one of the mechanisms underlying airway remodeling. This study aimed at identifying novel molecules involved in pediatric asthma-associated airway remodeling. Asthma model was established by challenging C57BL/6 mouse pups with ovalbumin (OVA). We found that the expression of Cadherin 11 (CDH11), a type II cadherin, was increased by OVA treatments in the airway epithelium. Our earlier microarray data suggested miRNA-451a-5p (miRNA-451a) as a potential regulator of CDH11. In contrast to CDH11, miRNA-451a expression decreased in the asthmatic lung. MiRNA-451a was then packaged into a lentivirus vector and systematically given to the asthmatic pups. Our data indicated that OVA-induced infiltration of inflammatory cells, including eosnophils, neutrophils, macrophages and lymphocytes, was reduced by miRNA-451a over-expression. EMT was initiated in asthmatic mice as demonstrated by increased alpha-smooth muscle actin (α-SMA) positive cells present in airway epithelium, which was inhibited by miRNA-451a. CDH11 elevation in vivo was also inhibited by miRNA-451a. Dual-Luciferase analysis further showed CDH11 as a novel valid target of miRNA-451a. Additionally, in vitro, EMT was triggered in human 16HBE airway epithelial cells by pro-fibrotic transforming growth factor β (TGF-β). Corresponding to the anti-EMT effects observed in vivo, miRNA-451a also inhibited TGF-β-induced collagen deposition in cultured airway epithelial cells by targeting in CDH11. In summary, our study demonstrates that the deregulated miRNA-451a-CDH11 axis contributes to airway remodeling in childhood asthma.

Topics: Airway Remodeling; Allergens; Animals; Animals, Newborn; Asthma; Cadherins; Cells, Cultured; Disease Models, Animal; Humans; Hypersensitivity; Male; Mice; Mice, Inbred C57BL; MicroRNAs; Ovalbumin; Respiratory Mucosa; Signal Transduction; Transforming Growth Factor beta

Prenatal particle exposure has been shown to increase allergic responses in offspring. Carbon nanotubes (CNTs) possess immunomodulatory properties, but it is unknown whether maternal exposure to CNTs interferes with offspring immune development. Here, C57Bl/6J female mice were intratracheally instilled with 67 of μg multiwalled CNTs on the day prior to mating. After weaning, tolerance and allergy responses were assessed in the offspring. Offspring of CNT-exposed (CNT offspring) and of sham-exposed dams (CTRL offspring) were intranasally exposed to ovalbumin (OVA) once weekly for 5 weeks to induce airway mucosal tolerance. Subsequent OVA sensitization and aerosol inhalation caused low or no OVA-specific IgE production and no inflammation. However, the CNT offspring presented with significantly lower OVA-specific IgG1 levels than CTRL offspring. In other groups of 5-week-old offspring, low-dose sensitization with OVA and subsequent OVA aerosol inhalation led to significantly lower OVA-specific IgG1 production in CNT compared to CTRL offspring. OVA-specific IgE and airway inflammation were non-significantly reduced in CNT offspring. The immunomodulatory effects of pre-gestational exposure to multiwalled CNTs were unexpected, but very consistent. The observations of suppressed antigen-specific IgG1 production may be of importance for infection or vaccination responses and warrant further investigation.

Topics: Animals; Antibody Formation; Antigens; Female; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin G; Inflammation; Maternal Exposure; Mice; Mice, Inbred BALB C; Nanotubes, Carbon; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects

We previously reported attenuation of serum OVA-specific IgE levels and of lymphocyte-derived IL-4, both nominal markers of allergic immunity, following injection of a combination of homologous (mouse) polyclonal anti-idiotypic immunoglobulin (Ig) and immune Ig in BALB/c mice. We predicted this might generalize to other species and using heterologous mixtures of Igs. This was assessed in mice using OVA sensitization in the presence of human Igs as a source of both anti-idiotype Ig and immune Ig and in dogs with peanut butter-induced allergic responses.. Eight-week-old BALB/c mice received OVA immunization and 5 weekly injections of immune Ig or anti-idiotype Ig from either homologous (mouse) or heterologous (human) sources. Five-month-old Beagles received weekly topical exposure (on the abdomen) to peanut butter and treatment with pooled dog Ig and dog antirabies immune Ig, or a combination of human IMIG and human anti-Tet. All mice/dogs thereafter received a final allergen challenge, and serum IgG, IgE, and allergen-induced IL-2/IL-4 and IL-31 production in 72 hr cultures was measured.. In mice attenuation of OVA-induced allergy (IgE-specific Ig and OVA-induced IL-4) was seen using both mouse and human Ig mixtures, without effect on OVA serum IgG or OVA-induced IL-2. Attenuation of concanavalin A- (ConA-) induced IL-4 : IL-2 production and of peanut butter-induced IL-4 and IL-31 was seen in dogs receiving combinations of both heterologous and homologous immune Igs and anti-idiotype Igs, with no decline in IL-2 production. Allergen-specific IgE/IgG was not detectable in dog serum, but there was a trend to lower total serum IgE levels (and decreased IgE : IgG ratios).. Homologous and heterologous combinations of polyclonal IMIG and immune Ig attenuate allergic responses in mice and dogs. This treatment protocol represents a novel approach which can be adapted for allergic desensitization in veterinary and human use.

Topics: Allergens; Animals; Antibodies, Anti-Idiotypic; Arachis; Desensitization, Immunologic; Dogs; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Peanut Oil

Angiotensin-II (Ang-II) receptor plays a role in allergic airway inflammation; however, the underlying mechanism and role of macrophages need better understanding. In the present study, angiotensin-II infusion (1 μg/kg/min) in ovalbumin-induced airway inflammation mice model significantly decreased immune cell infiltration, goblet cell hyperplasia, and eosinophil numbers in lungs. Ang-II infusion increased M1 and decreased M2 macrophage population in bronchoalveolar lavage fluid and respective macrophage markers in lung macrophages. Similarly, in vitro Ang-II treatment in murine bone marrow-derived macrophages (BMDMs) induced M1 and reduced M2 macrophage phenotype with enhanced bactericidal activity. Mechanistically, Ang-II inhibits Let-7c and miR-99a expression in BMDMs and in vivo as well. Lentiviral overexpression of Let-7c and miR-99a miRNAs in BMDMs abrogated Ang-II-induced M1 phenotype activation and promoted M2 phenotype, which is governed by targeting TNFα by miR-99a. In lung macrophages, ovalbumin-induced TNFα inhibition was rescued after Ang-II treatment. In BMDMs, knockdown of TNFα abrogated Ang-II-induced M2 to M1 macrophage phenotype switch and associated bactericidal activity. Ang-II affects mature miRNA formation by enhancing Lin28B levels in macrophages in vivo and in vitro. Furthermore, Lin28B knockdown prevented Ang-II-mediated inhibition of mature Let-7c/miR-99a miRNA formation, M2 to M1 macrophage phenotype switch, and increased bactericidal activity. Therefore, present study suggests a role of Lin28B in Ang-II-induced Let-7c/miR-99a miRNA formation that consequently affects TNFα production, M1 phenotype activation, and allergic airway inflammation. Graphical Abstract Ovalbumin inhibits LIN28B expression thereby fails to inhibit premature to mature Let-7c/miR-99a miRNA formation. Mature miR-99a miRNA that inhibits TNFα consequently promotes M2 polarization and allergic airway inflammation. While Ang-II induces Lin28B, which inhibits Let-7c/miR-99a miRNA processing and mature miRNA formation, this results in increased TNFα levels that lead to M1 polarization and allergic airway inflammation inhibition.

Topics: Angiotensin II; Animals; Cell Polarity; Cells, Cultured; HEK293 Cells; Humans; Hypersensitivity; Macrophages, Alveolar; Mice; Mice, Inbred C57BL; MicroRNAs; Ovalbumin; Pneumonia; RNA-Binding Proteins

Allergic asthma is one of the most common chronic diseases of the airways, however it still remains underdiagnosed and hence undertreated. Therefore, an allergic asthma rat model would be useful to be applied in future therapeutic strategy studies. The aim of the present study was to develop an objective model of allergic asthma in atopic rats that allows the induction and quantification of anaphylactic shock with quantitative variables. Female Brown Norway rats were intraperitoneally sensitized with ovalbumin (OVA), alum and

Topics: Administration, Intranasal; Allergens; Animals; Asthma; Body Temperature; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Epithelial Cells; Female; Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Leukotrienes; Lung; Ovalbumin; Rats

Numerous studies indicate that toll-like receptor 2 (TLR2) led to divergent effects in asthma. The occurrence of autophagy in asthma pathogenesis is still incompletely understood. Here, we aimed to investigate the role of TLR2 and the underlying mechanisms in allergic airway inflammation and autophagy activation.. C57BL/6 and TLR2 knockout (TLR2. TLR2 expression was increased upon OVA challenge, and TLR2 deficiency was associated with decreased allergic airway inflammation. Meanwhile, TLR2 deficiency weakened autophagy activation. Moreover, inhibition of c-Jun N-terminal kinase (JNK) by SP600125 also suppressed OVA-induced allergic airway inflammation and autophagy activation. Interestingly, treating TLR2. TLR2 might contribute to the maintenance of allergic airway inflammation through JNK signaling pathway accompanying with autophagy activation. These findings may provide a novel signal target for prevention of allergic airway inflammation.

Topics: Animals; Autophagy; Disease Models, Animal; Goblet Cells; Hypersensitivity; Immunoglobulin E; Lung; MAP Kinase Signaling System; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Pneumonia; Proto-Oncogene Proteins c-akt; Toll-Like Receptor 2

Sublingual immunotherapy (SLIT) was introduced to deliver allergens in an effective and non-invasive route, which can be considered as an alternative for allergen-specific subcutaneous immunotherapy (SCIT). On the other hand, the use of gold nanoparticles (AuNPs) in allergen delivery has beneficial effects on sublingual immunotherapy. In addition, the molecular targeting agents like aptamers (Apt), have been widely applied for targeted drug delivery. Therefore, the current study aimed to evaluate the effects of dendritic cells (DCs)-specific Aptamer-modified AuNPs coated with ovalbumin (OVA) on the improvement of the SLIT outcome in the mouse model of allergy.. AuNPs with approximately 15 nm diameter were prepared by citrate reduction of HAuCl4. Afterward, Apt-modified AuNP complex was prepared and OVA was then loaded onto this complex. Following sensitization of Balb/c mice to OVA, SLIT was performed with Apt-AuNPs containing 5 µg OVA twice a week for a 2-month period. Allergen-specific IgE in serum, as well as cytokines secretion of spleen cells, were analyzed using ELISA. Also, nasopharyngeal lavage Fluid (NALF) was collected for total and eosinophil counts. Moreover, the lungs were removed for histopathological examination.. SLIT with Apt-modified AuNPs complex containing 5 μg OVA, decreased the IgE levels compared to the other groups. Also, IL-4 production has significantly decreased in spleen cells, while TGF-β and IFN-γ have significantly increased. The assessment of NALF in the group treated by this complex showed a decrease in total cell as well as in eosinophil count. Also, the examination of lung tissues revealed that, in the group treated by this complex, inflammation and perivascular infiltration were lesser than the other groups, which were observed in only one vessel of tissue.. It was shown that, Sublingual immunotherapy with DC specific Apt-modified AuNPs containing 5 μg OVA can improve the Th1 and Treg immunomodulatory responses.

Topics: Allergens; Animals; Aptamers, Nucleotide; Biocompatible Materials; Cytokines; Dendritic Cells; Disease Models, Animal; Drug Carriers; Eosinophils; Female; Gold; Hypersensitivity; Immunoglobulin E; Lung; Metal Nanoparticles; Mice, Inbred BALB C; Nasal Lavage Fluid; Ovalbumin; Spleen; Sublingual Immunotherapy; T-Lymphocytes, Regulatory; Th1 Cells

The enhanced type 2 helper (Th2) immune response is responsible for the pathogenesis of allergic asthma. To suppress the enhanced Th2 immune response, activation of the Th1 immune response has been an alternative strategy for anti-asthma therapy. In this context, effective Th1-inducing adjuvants that inhibit the development of allergic asthma but do not flare the side effects of the primary agent are required in clinical treatment and preventive medicine.. In this study, we aimed to determine the regulation of the Th2 type immune response in asthma by a novel immunostimulatory oligodeoxynucleotide (ODN) derived from Cryptococcus neoformans, termed ODN112, which contains a cytosine-guanine (CG) sequence but not canonical CpG motifs.. Using an ovalbumin-induced asthma mouse model, we assessed the effect of ODN112 on prototypical asthma-related features in the lung and on the Th1/Th2 profile in the lymph nodes and lung of mice treated with ODN112 during sensitization.. ODN112 treatment attenuated asthma features in mice. In the bronchial lymph nodes of the lungs and in the spleen, ODN112 increased interferon-γ production and attenuated Th2 recall responses. In dendritic cells (DCs) after allergen sensitization, ODN112 enhanced cluster of differentiation (CD) 40 and CD80 expression but did not alter CD86 expression. Interleukin-12p40 production from DCs was also increased in a Th2-polarizing condition. Our results suggest that ODN112 is a potential Th1-inducing adjuvant during Th2 cell differentiation in the sensitization phase.

Topics: Allergens; Animals; Asthma; Cell Differentiation; CpG Islands; Cryptococcus neoformans; Dendritic Cells; Disease Models, Animal; Female; Humans; Hypersensitivity; Mice; Mice, Inbred C57BL; Oligodeoxyribonucleotides; Ovalbumin; Th1-Th2 Balance; Th2 Cells; Toll-Like Receptor 9

Asthma is a serious global health problem, severely affecting the lives of sufferers and their families. An exceptionally hygienic home and reduced microbial exposure can aggravate the incidence of childhood asthma.. Specific-pathogen-free BALB/c mice were pre-treated with bacterial lysate (BL; 1 mg/kg) as a high microbial load maternal mouse model, and then, the offspring mice were established as an allergic airway disease (AAD) model. The expression levels of TLR2, TLR4, and HDAC9 in the mother's intestine and the offspring's lungs were detected. Relevant indicators of regulatory T cells (Tregs) were identified in the mother and offspring mice. The changes in the expression of Th1-, Th2-, Th9-, and Th17-related cytokines in the offspring mice were evaluated among different pre-treated groups.. After augmenting the mothers' intestinal microbiota through oral BL gavage, the expression of TLR2 and TLR4 in the colon mucosa and colon lymphoid tissues was enhanced and that of HDAC9 in the colon mucosa was decreased, and the proportion of spleen Tregs was increased. The offspring showed similar changes in the AAD model compared with the offspring of the control-group mothers: TLR2 and TLR4 expression in the lungs and the proportion of spleen Tregs increased, HDAC9 expression in the lungs decreased, and AAD-induced airway pathologic characteristics were reversed; additionally, Th1/Th2 and Th9 imbalances were rectified.. This study presents a new framework for the prevention of childhood asthma, elucidating the mechanism of regulating the mother's intestinal microbiome to protect the offspring's early asthma via animal experiments.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Humans; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Microbiota; Ovalbumin; Th2 Cells

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Ovalbumin; Oxidative Stress; Plant Exudates; Plant Stems; Sambucus nigra; Spleen; Th2 Cells

Subcutaneous allergen-specific immunotherapy (SCIT) qualifies as a promising approach for the permanent cure of IgE-mediated airway allergies, which can often manifest into allergic rhinitis and other allergic respiratory diseases. SCIT entails repeated administration of a high allergen dose into the subcutaneous (sc) region using a hypodermic needle for many (3-5) years, which is inconvenient and painful and reduces patient compliance. To overcome these limitations, we hypothesized that microneedles (MNs), which are minimally invasive and painless, could provide a novel approach for allergen desensitization by depositing the allergen into the superficial layers of the skin. To test this hypothesis, we compared MNs and SCIT for allergen desensitization in a mouse model of ovalbumin (Ova)-induced airway allergy. Mice were first made allergic to Ova and then treated with MNs coated with Ova (with or without CpG as an adjuvant) or via SCIT-Ova + alum (subcutaneous Ova + alum injections) for comparison. Treatment with coated MNs significantly induced Ova-specific serum IgG antibodies in a manner comparable to SCIT-Ova + alum-treated group. To test the efficacy against allergen challenge, treated mice were challenged with Ova via the nasal route. Coated MNs with Ova and CpG (MN-Ova + CpG) considerably suppressed the airway inflammation in allergic mice, evidenced by downregulation of proinflammatory cytokines (IL-5 and IL-13), upregulation of anti-inflammatory cytokine IL-10, and activation of Ova-specific immune response in bronchoalveolar (BAL) fluid. The therapeutic capacity of MN-based allergy treatment was further validated by the reduction in eosinophil and mast cell infiltration in the lung tissues of mice treated with MN-Ova + CpG, and low deposition of mucus inside their lung bronchioles. Overall, coated MNs ameliorated the symptoms of airway allergy in mice similar to SCIT and could provide a novel means of painless allergen-specific immunotherapy.

Topics: Adjuvants, Immunologic; Administration, Cutaneous; Allergens; Alum Compounds; Animals; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunoglobulin G; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Needles; Oligodeoxyribonucleotides; Ovalbumin

Numerous data obtained by different research laboratories indicate that specific IgE production is triggered independently of specific IgG or IgA ones and so it is not linked to fully matured germinal centers formation in the secondary lymphoid organs. The aim of this study was to clarify whether specific IgE production is triggered by low antigen doses administrated in tertiary tissues enriched by lymphoid structures.. Ovalbumin (OVA) in different doses (100 ng to 10 μg) was administrated three times a week for 4-5 weeks intraperitoneally (i.p.) or subcutaneously (s.c.) to female BALB/c mice in the wither region which is enriched in fat-associated lymphoid clusters or in the foot pad region not containing them.. OVA-specific IgE was predominantly induced by low but not high antigen doses and only after immunization into the withers. IgE isotype switching was triggered exclusively in the withers adipose tissue but not in the regional lymph nodes while mature IgE expressing cells were observed both in the withers and lymph nodes. Anti-proliferative genotoxic stress inducing drugs shifted the balance from IgG1 towards IgE production.. Tertiary lymphoid structures possess unique environment where B-cell antibody isotype switching to IgE predominantly occurs. This phenomenon is partially explained by hampered proliferation of B-cells in these structures.

Topics: Allergens; Animals; B-Lymphocytes; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Female; Humans; Hypersensitivity; Immunoglobulin Class Switching; Immunoglobulin E; Immunoglobulin G; Lymphoid Tissue; Mice; Ovalbumin; Tertiary Lymphoid Structures

Asthma is a chronic respiratory disease orchestrated by immune and structural cells. Identification of novel therapeutic strategies are needed for asthma due to the limitations of existing therapies. We have validated the anti-inflammatory, anti-asthmatic and immunomodulatory therapeutic properties of herbal decoction, Divya-Swasari-Kwath (DSK) using mouse model of ovalbumin (OVA) induced allergic asthma.. HPLC analysis identified the presence of Rutin, Glycyrrchzin, Gallic acid, Cinnamic acid, Chlorogenic acid, Caffeic acid and Piperine as bioactive herbal metabolites in DSK. Therapeutic treatment with herbal decoction DSK significantly alleviated the pathological features of allergic asthma including inflammatory cell accumulation in Broncho-Alveolar Lavage (BAL) fluids, specifically lymphocytes and eosinophils, lung inflammation, oxidative stress, airway remodelling, and pro-inflammatory cytokine levels. H&E analysis of lung tissue sections identified attenuated inflammatory cell infiltration and thickening of bronchial epithelium by DSK. PAS staining and MT staining identified decrease in OVA-induced mucus hyper secretion and peri-bronchial collagen deposition respectively, upon DSK treatment. Treatment with DSK increased the mRNA expression of antioxidative defence gene Nrf-2 and its downstream target genes HO-1 and NQO-1. In the same line, biochemical analysis for the markers of oxidative/antioxidant system confirmed the restoration of activity of Catalase, GPx, SOD and EPO and the levels of GSH, GSSG, MDA and Nitrite in whole lungs. In line with PAS staining, DSK treatment decreased the OVA-induced expression of Muc5AC and Muc5B genes. DSK treatment reduced the steady state mRNA expression levels of IL-6, IL-1β, TNF-α, IL-4, -5, -33, IFN-γ in whole lung; and IL-6, TNF-α and IL-1β protein levels in BALF.. Collectively, our results suggest that herbal decoction DSK is effective in protecting against allergic airway inflammation and remodelling by regulating anti-oxidant mechanisms. We postulate that DSK could be the potential therapeutic option for allergic asthma management.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunologic Factors; Lung; Male; Medicine, Ayurvedic; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin; Oxidative Stress; Plant Preparations; Pneumonia

Airway dendritic cells (DCs) are recognized as important factors in the mechanisms of allergic inflammatory diseases. Suppressor of cytokine signaling 3 (SOCS3) is involved in regulating the functions of T cells and macrophages, but the roles of SOCS3-expressing DCs in the pathogeneses of allergic inflammatory diseases are still controversial. We compared the effects of adoptively transferred SOCS3

Topics: Adoptive Transfer; Animals; Asthma; Bone Marrow Cells; Cell Movement; Cell Proliferation; Chemotaxis; Cytokines; Dendritic Cells; Hypersensitivity; Inflammation; Injections, Intraperitoneal; Lipopolysaccharides; Lung; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Signal Transduction; STAT Transcription Factors; Suppressor of Cytokine Signaling 3 Protein; T-Lymphocytes

Asthma, one of the significant public health problems, is triggered by certain inflammatory processes in the airways that are not addressed propitiously by current therapies. Though pieces of evidence on allergic asthma mitigation by the anti-inflammatory bioflavonoid chrysin (CHR) are accumulating, poor bioavailability, and low solubility curtail drug development. To overcome these shortcomings, CHR loaded nanoparticle (CHR-NP) was formulated, and its salutary effect in preclinical murine allergic asthma model via the peroral route was evaluated. The spherical nanosized particles showed slow, sustained release in vitro. Moreover, CHR-NP dramatically reduced the serum IgE, ovalbumin (OVA)-induced lung histological alteration, as well as Th2 (T-helper 2) cytokines in the bronchoalveolar lavage fluid (BALF). It also suppressed the elevated serum pro-inflammatory cytokines and their upstream TLR/NF-κB/NLRP3 pathway activation in lung superior to CHR and almost identical to dexamethasone (DEX). Thus this study suggests the potentiality of CHR-NP in ameliorating allergic asthma progression.

Topics: A549 Cells; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Flavonoids; Humans; Hypersensitivity; Immunoglobulin E; Inflammation Mediators; Lung; Male; Mice; Mice, Inbred BALB C; Microscopy, Atomic Force; Microscopy, Electron, Transmission; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Polylactic Acid-Polyglycolic Acid Copolymer; Toll-Like Receptors

Topics: Animals; Anti-Inflammatory Agents; Asthma; Dexamethasone; Disease Models, Animal; Drug Carriers; Hydrogen Peroxide; Hypersensitivity; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin

In allergic bronchial asthma, an increased smooth muscle contractility of the airways is one of the causes of the airway hyperresponsiveness (AHR). Increasing evidence also suggests a possible involvement of microRNAs (miRNAs) in airway diseases, including asthma, although their roles in function and pathology largely unknown. The current study aimed to determine the role of a miRNA, miR-140-3p, in the control of protein expression of CD38, which is believed to regulate the contraction of smooth muscles, including the airways. In bronchial smooth muscles (BSMs) of the mice that were actively sensitized and repeatedly challenged with ovalbumin antigen, an upregulation of CD38 protein concurrently with a significant reduction of miR-140-3p was observed. In cultured human BSM cells (hBSMCs), transfection with a synthetic miR-140-3p inhibitor caused an increase in CD38 protein, indicating that its basal protein expression is regulated by endogenous miR-140-3p. Treatment of the hBSMCs with interleukin-13 (IL-13), an asthma-related cytokine, caused both an upregulation of CD38 protein and a downregulation of miR-140-3p. Transfection of the hBSMCs with miR-140-3p mimic inhibited the CD38 protein upregulation induced by IL-13. On the other hand, neither a CD38 product cyclic ADP-ribose (cADPR) nor its antagonist 8-bromo-cADPR had an effect on the BSM contraction even in the antigen-challenged mice. Taken together, the current findings suggest that the downregulation of miR-140-3p induced by IL-13 might cause an upregulation of CD38 protein in BSM cells of the disease, although functional and pathological roles of the upregulated CD38 are still unclear.

Topics: ADP-ribosyl Cyclase 1; Animals; Asthma; Bronchi; Cell Line; Disease Models, Animal; Gene Expression Regulation; Humans; Hypersensitivity; Interleukin-13; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin

Homeostatic mucosal immune responses are fine-tuned by naturally evolved interactions with native microbes, and integrating these relationships into experimental models can provide new insights into human diseases. Here, we leverage a murine-adapted airway microbe, Bordetella pseudohinzii (Bph), to investigate how chronic colonization impacts mucosal immunity and the development of allergic airway inflammation (AAI). Colonization with Bph induces the differentiation of interleukin-17A (IL-17A)-secreting T-helper cells that aid in controlling bacterial abundance. Bph colonization protects from AAI and is associated with increased production of secretory leukocyte protease inhibitor (SLPI), an antimicrobial peptide with anti-inflammatory properties. These findings are additionally supported by clinical data showing that higher levels of upper respiratory SLPI correlate both with greater asthma control and the presence of Haemophilus, a bacterial genus associated with AAI. We propose that SLPI could be used as a biomarker of beneficial host-commensal relationships in the airway.

Topics: A549 Cells; Adolescent; Adult; Animals; Antigens; Bordetella; Child; Colony Count, Microbial; Disease Models, Animal; Host Microbial Interactions; Humans; Hypersensitivity; Immunity; Inflammation; Lung; Mice, Inbred C57BL; Microbiota; Ovalbumin; Secretory Leukocyte Peptidase Inhibitor; Th17 Cells; Transcriptome; Young Adult

The superantigen

Topics: Animals; Biomarkers; Cytokines; Disease Models, Animal; Enterotoxins; Female; Hypersensitivity; Immunization; Immunoglobulin E; Immunomodulation; Leukocytes; Lymphocyte Activation; Lymphocytes; Mice; Ovalbumin; Staphylococcal Infections; Staphylococcus aureus; Superantigens

While sublingual immunotherapy (SLIT) is known as an allergen-specific treatment for type-1 allergies, how it controls allergic pathogenesis remains unclear. Here, we show the prerequisite role of conventional dendritic cells in submandibular lymph nodes (ManLNs) in the effectiveness of SLIT for the treatment of allergic disorders in mice. Deficiency of conventional dendritic cells or CD4

Topics: Animals; Antibody Specificity; CD4 Antigens; Dendritic Cells; Forkhead Transcription Factors; Hypersensitivity; Immunity, Cellular; Immunization; Immunoglobulins; Immunotherapy; Lymph Nodes; Mice; Ovalbumin; T-Lymphocytes, Regulatory

We established a mouse model of allergic asthma by sensitizing with chicken ovalbumin. The volatile oils and decoctions from raw, wine- and vinegar-steamed Schisandra chinensis fruits were intragastrically administrated to the mice. Atomization, serum IgE, IL-2, IL-4 and IFN-γ in the lung homogenates and pathological sections were evaluated to compare the effect of these volatile oils and decoctions on allergic asthma in mice. The results showed that all Schisandra volatile oils could significantly suppress allergic asthma in mice. Raw Schisandra volatile oil was most effective followed by volatile oils extracted from wine-steamed and vinegar-steamed Schisandra. The decoctions had no significant impact. Our findings demonstrated that volatile oil was the active ingredient in Schisandra, and raw Schisandra could be used to prevent cough and asthma.

Topics: Animals; Asthma; Cyclooctanes; Fruit; Hypersensitivity; Lignans; Male; Mice; Oils, Volatile; Ovalbumin; Plant Extracts; Polycyclic Compounds; Schisandra

The hygiene hypothesis suggests a link between parasitic infections and immune disorders, such as allergic diseases. We previously showed that infection with. Mice were intranasally treated with TLA either i) prior to sensitization, ii) during sensitization and challenge, or iii) after sensitization with ovalbumin (OVA). Recruitment of inflammatory cells to the lung, cytokine levels in restimulated lung and spleen cell cultures as well as levels of OVA-specific antibodies in serum were measured. In parallel, the effect of native TLA, heat-inactivated (hiTLA) or deglycosylated TLA (dgTLA) on sensitized splenocytes was evaluated. When applied together with OVA i) during systemic sensitization and local challenge or ii) exclusively during local challenge, TLA reduced infiltration of eosinophils into the lung, OVA-specific type 2 cytokines in restimulated lung cell cultures, and partially, type 2 cytokines in restimulated spleen cell cultures in comparison to allergic controls. No beneficial effect was observed when TLA was applied prior to the start of sensitization. Analysis of epitope sugars on TLA indicated a high abundance of mannose, fucose, N-acetylglucosamine, and N-acetylgalactosamine. Deglycosylation of TLA, but not heat-inactivation, abolished the potential of TLA to reduce type 2 responses. We showed that mucosal application of TLA reduced the development of experimental allergy in mice. The beneficial effects depended on the timing of the application in relation to the time point of sensitization. Not only co-application, but also therapy in sensitized/allergic animals with native TLA reduced local allergic responses. Furthermore, we show that TLA is highly glycosylated and glycoconjugates seem to play a role in anti-allergic effects. In summary, given the powerful modulatory effect that TLA exhibits, understanding its exact mechanisms of action may lead to the development of novel immunomodulators in clinical application.

Topics: Allergens; Animals; Antigens, Protozoan; Carbohydrates; Cell Line; Chlorocebus aethiops; Cytokines; Female; Hypersensitivity; Immunologic Factors; Lung; Lung Diseases; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Mucosa; Spleen; Toxoplasma; Toxoplasmosis; Vero Cells

Topics: Animals; Anti-Allergic Agents; Antrodia; Benzodioxoles; Cells, Cultured; Cytokines; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Hypersensitivity; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Th2 Cells

Helminths and their products can shape immune responses by modulating immune cells, which are dysfunctional in inflammatory diseases such as asthma. We previously identified SJMHE1, a small molecule peptide from the HSP60 protein of Schistosoma japonicum. SJMHE1 can inhibit delayed-type hypersensitivity and collagen-induced arthritis in mice. In the present study, we evaluated this peptide's potential intervention effect and mechanism on ovalbumin-induced asthma in mice. SJMHE1 treatment suppressed airway inflammation in allergic mice, decreased the infiltrating inflammatory cells in the lungs and bronchoalveolar lavage fluid, modulated the production of pro-inflammatory and anti-inflammatory cytokines in the splenocytes and lungs of allergic mice, reduced the percentage of Th2 cells and increased the proportion of Th1 and regulatory T cells (Tregs). At the same time, Foxp3 and T-bet expression increased, and GATA3 and RORγt decreased in the lungs of allergic mice. We proved that SJMHE1 can interrupt the development of asthma by diminishing airway inflammation in mice. The down-regulation of Th2 response and the up-regulation of Th1 and Tregs response may contribute to the protection induced by SJMHE1 in allergic mice. SJMHE1 can serve as a novel therapy for asthma and other allergic or inflammatory diseases.

Topics: Animals; Asthma; Cytokines; Forkhead Transcription Factors; GATA3 Transcription Factor; Gene Expression Regulation; Hypersensitivity; Inflammation; Lung; Lymphocyte Subsets; Mice; Mice, Inbred BALB C; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Peptides; RNA, Messenger; Schistosoma japonicum; Spleen; T-Box Domain Proteins

Although the bulk of research into the biology of serotonin 5-HT. An 18-week ovalbumin challenge period was performed to generate persistent, chronic asthma in BALB/c mice. Four once daily intranasal treatments of (R)-DOI were administered one week after allergen cessation, with respiratory parameters being measured by whole-body plethysmography (WBP). Cytokine and chemokine levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) in homogenized lung tissue, bronchoalveolar (BALF) fluid was analyzed for chemokine modulation by multiplex assays, and Periodic Acid-Schiff and Masson's Trichrome staining was performed to determine goblet cell infiltration and overall changes to lung morphology.. 5-HT. Overall, these data provide support for the therapeutic potential of (R)-DOI and 5-HT

Topics: Airway Remodeling; Amphetamines; Animals; Asthma; Chronic Disease; Female; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Receptors, Serotonin, 5-HT2; Serotonin 5-HT2 Receptor Agonists

Allergic asthma is a chronic inflammatory disease and involves many cells and cellular components. Cataract is a condition that affects the transparency of the lens, which the opacity of the lens caused by any innate or acquired factor degrades its transparency or changes in color. During the establishment of asthma model of rats with chicken ovalbumin nebulization, it was found that asthmatic rats were more likely to have monocular or binocular cataract symptoms than normal rats. Considering that they are all induced by immune imbalance, inflammation, etc., there may be some correlation in the mechanism, and many clues showed that both diseases are associated with activation of the PI3K-AKT-mTOR signaling pathway. Therefore, we hypothesized that the PI3K-AKT-mTOR signaling pathway produces inflammatory or immune imbalance based on allergy leading to cataract.

Topics: Animals; Asthma; Cataract; Disease Models, Animal; Hypersensitivity; Inflammation; Male; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; TOR Serine-Threonine Kinases

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Immunophenotyping; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Nickel; Ovalbumin; Particle Size

Treatment of patients with cat allergy with peptides derived from Fel d 1 (the major cat allergen) ameliorated symptoms of cat allergy in phase 2 clinical trials.. We sought to demonstrate that the tolerance induced by Fel d 1 peptide immunotherapy can be exploited to reduce allergic responses to a second allergen, ovalbumin (OVA), in mice sensitized dually to OVA and Fel d 1.. Induction of tolerance to OVA was achieved through simultaneous exposure to both allergens after peptide treatment. Functional tolerance to each allergen was assessed in a model of allergic airways disease in which treated mice were protected from eosinophilia, goblet cell hyperplasia, and T. Suppression of allergic responses to cat allergen challenge was associated with significant increases in numbers of CD4. These observations suggest that immune tolerance induced by peptide immunotherapy can be used experimentally to treat an allergic response to another allergen and that the molecular mechanisms underlying induction of tolerance to a treatment-specific allergen and a bystander allergen might be different.

Topics: Allergens; Animals; B-Lymphocytes; Bystander Effect; Cytokines; Desensitization, Immunologic; Female; Glycoproteins; Hypersensitivity; Immune Tolerance; Lung; Mice, Inbred BALB C; Ovalbumin; Peptides; T-Lymphocytes

Serum IL-22 levels are increased in patients with atopic dermatitis, which commonly precedes asthma in the atopic march. Epicutaneous sensitization in mice results in T. We sought to determine the role of IL-22 in antigen-driven airway allergic inflammation after inhalation challenge in epicutaneously sensitized mice.. Wild-type (WT) and Il22. Epicutaneous sensitization preferentially elicited an IL-22 response compared with intraperitoneal immunization. Intranasal challenge of mice epicutaneously sensitized with OVA elicited in the lungs Il22 mRNA expression, IL-22 production, and accumulation of CD3. Epicutaneous sensitization promotes generation of antigen-specific IL-22-producing T cells that promote airway inflammation and AHR after antigen challenge, suggesting that IL-22 plays an important role in the atopic march.

Topics: Allergens; Animals; Asthma; Dermatitis, Atopic; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Inflammation; Interleukin-22; Interleukins; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Respiratory Hypersensitivity; Skin; Th2 Cells

Severe, chronic eye allergy is an understudied, vision-threatening condition. Treatments remain limited. We used a mouse model of severe allergic eye disease (AED) to determine whether topical application of the pro-resolution mediator Resolvin D1 (RvD1) terminates the response. AED was induced by injection of ovalbumin (OVA) followed by topical challenge of OVA daily. RvD1 was applied topically prior to OVA. Clinical symptoms were scored. Eye washes were assayed for MUC5AC. After 7 days, eyes were removed and the number of goblet cells, T helper cell responses and presence of immune cells in draining lymph nodes and conjunctiva determined. Topical RvD1 treatment significantly reduced symptoms of AED. RvD1 did not alter the systemic type 2 immune response in the lymph nodes. AED increased the total amount of goblet cell mucin secretion, but not the number of goblet cells. RvD1 prevented this increase, but did not alter goblet cell number. Absolute numbers of CD4 + T cells, total CD11b + myeloid cells, eosinophils, neutrophils, and monocytes, but not macrophages increased in AED versus RvD1-treated mice. We conclude that topical application of RvD1 reduced the ocular allergic response by local actions in conjunctival immune response and a decrease in goblet cell mucin secretion.

Topics: Allergens; Animals; Cells, Cultured; Chronic Disease; Disease Models, Animal; Docosahexaenoic Acids; Eye Diseases; Goblet Cells; Humans; Hypersensitivity; Immunity, Cellular; Mice; Mice, Inbred C57BL; Mucin 5AC; Ovalbumin

Mast cell-mediated allergic diseases are a significant global health problem. Nitric oxide (NO) produced by acute type 1 allergies greatly suppresses hepatic cytochrome P450 (CYP) metabolism. A recent in vitro study demonstrated that repeated FcεRI-mediated activation intrinsically modulates mast cell function. We investigated the effect of ovalbumin (OVA) challenges on CYP activity and NO production under real immune responses.. After repeated sensitization with OVA once a week, serum nitrate plus nitrite (NOx) and total plasma immunoglobulin E concentrations were measured using commercially available kits. Hepatic microsomal CYP-specific activities and protein expression were determined using typical substrates and by western blot, respectively. In the liver, the levels of inducible NO synthase (iNOS), F4/80, and c-kit mRNA were determined by real-time polymerase chain reaction. Hepatic total NOS activity was measured using a colorimetric assay kit.. When mice received multiple OVA challenges, the 11th sensitization elevated NOx concentrations in serum and suppressed the activities of five major CYPs without altering protein expression levels. After the 7th, 11th, and 15th sensitizations, F4/80-positive Kupffer cell and hepatic c-kit-dependent mast cell mRNA levels were similar to those of the control. The 7th and 11th sensitizations increased hepatic iNOS mRNA expression to 15-fold and threefold above control levels, respectively, but did not enhance the total NOS activity in the liver.. Multiple OVA challenges, unlike acute sensitization, greatly reduced serum NOx levels. The challenge-suppressed hepatic CYP metabolism was likely related to the increased serum NOx. Serum NOx may be an endogenous marker for CYP metabolism inhibition in type 1 allergic diseases.

Topics: Animals; Cytochrome P-450 Enzyme System; Female; Hypersensitivity; Immunoglobulin E; Liver; Mast Cells; Mice, Inbred ICR; Nitric Oxide; Nitric Oxide Synthase Type II; Ovalbumin; Real-Time Polymerase Chain Reaction

Topics: Adaptive Immunity; Allergens; Animals; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Humans; Hypersensitivity; Immunoglobulin E; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; OX40 Ligand; Peptides; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; STAT6 Transcription Factor; Th2 Cells

Rush desensitization can provide short-term tolerance to individuals who are allergic to certain medications in instances where other therapeutic interventions are limited. While rush desensitization (DS) is typically successful in preventing adverse type I hypersensitivity reactions, the mechanism of allergic protection remains unknown. Given the rise in prevalence of individuals displaying multiple allergies, understanding the impact of rush DS on "bystander" allergens, or additional allergens to which an individual is sensitized, could help inform clinical recommendations.. To evaluate the effect of rush DS on bystander sensitization.. We used a murine model of rush DS, whereby BALB/c mice were sensitized to ovalbumin (OVA) and desensitized through repeated intraperitoneal injections of OVA. Using a local anaphylaxis assay, we measured ear swelling by Evans blue extravasation following intradermal challenge. In studies to measure the impact on bystander antigens, a modified protocol was used in which mice were dually sensitized to OVA and Keyhole limpet hemocyanin (KLH), and densensitized to either OVA or KLH prior to allergic challenge.. The immunological effects of rush DS were independent of changes in Th1 and Th2 cytokine production and circulating OVA-IgE levels. Instead, rush DS resulted in subclinical degranulation of mast cells prior to challenge. In our dual sensitization model, rush DS with a single antigen conferred protection against allergic challenge to a secondary antigen. Bystander protection required prior sensitization, as DS with an irrelevant antigen did not impact allergic responsiveness.. We reveal that a key mechanism of rush DS protection against allergic responsiveness may be the subclinical degranulation of mast cells. Therefore, performing rush DS to a single antigen to which one is IgE-sensitized may be sufficient to desensitize to multiple allergens. Future studies could lead to streamlined protocols of rush DS for patients with multiple allergies.

Topics: Allergens; Anaphylaxis; Animals; Antigens; Biomarkers; Cell Degranulation; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin E; Mast Cells; Mice; Ovalbumin; T-Lymphocyte Subsets

A reverse targeting drug delivery based on antigen-modified nanoparticles provided an innovative strategy for effectively alleviating or inhibiting immune response. In this study, a dual fluorescent reverse targeting drug delivery system based on curcumin-loaded ovalbumin nanoparticles is developed for allergy treatment. The self-crosslinked ovalbumin nanoparticles achieved the double function of reverse targeting and sustained delivery carriers to maximize the anti-allergy of curcumin. Using a murine model of ovalbumin-induced allergy, this drug delivery system suppressed antigen-specific IgG1 and IgE production, inhibited CD4

Topics: Animals; Curcumin; Drug Delivery Systems; Hypersensitivity; Mice; Nanoparticles; Ovalbumin

Clinical evidence gathered in Chinese communities suggested that acupoint sticking therapy could be an alternative treatment for asthma-related diseases. However, its underlying mechanism is still poorly understood.. In this study, we aimed to investigate the mechanism of the anti-inflammatory effect of acupoint sticking application with 'Treatment of Winter Disease in Summer' (TWDS) prescription by using metabolomics.. Allergic asthma in guinea pig was sensitized and challenged by ovalbumin (OVA). Histopathological evaluation of the lung tissue was performed by hematoxylin and eosin (H&E) staining and Masson's trichrome staining. The levels of Th2 cytokine and IgE level in serum were measured using enzyme-linked immunoassay (ELISA). The mRNA expression levels of IL-4, IL-5, IL-13 and orosomucoid-like 3 (ORMDL3) were measured using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Proteins of NF-κB signaling pathway were measured using western blot. The serum metabolomics profiles were obtained by using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS).. The overall results confirmed that AST with TWDS prescription had a significant protective effect against OVA-induced allergic asthma in guinea pig. This treatment not only attenuated airway inflammation and collagen deposition in the airway, but also decreased the levels of IL-4, IL-5, IL-13 and IgE in serum. In addition, metabolomics results indicated that metabolisms of phospholipid, sphingolipid, purine, amino acid and level of epinephrine were restored back to the normal control level. Moreover, results of the gene expression of ORMDL3 in lung tissues indicated that AST using TWDS could alter the sphingolipid metabolism. Further western blotting analysis also showed that its anti-inflammatory mechanism was by decreasing the phosphorylation of p65 and IκB.. The study demonstrated that metabolomics provides a better understanding of the actions of TWDS acupoint sticking therapy on OVA-induced allergic asthma.

Topics: Acupuncture Therapy; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Drugs, Chinese Herbal; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Lung; Male; Membrane Proteins; Metabolomics; NF-kappa B; Ovalbumin; Signal Transduction

micro-RNAs (miRNAs) are non-coding RNAs which play important role in human diseases. Dysregulated miRNAs have been identified in asthma patient while their precise roles in asthma are not well elucidated. We compared the expression level of total 11 miRNAs between PMA/A23187-treated and control HMC-1 mast cells. We determined the effect of miR-20a on inflammation by overexpressing miR-20a mimic or its antagonist. We further predicted histone deacetylase 4 (HDAC4) as potential target of miR-20a and explored the effects of miR-20a on HDAC4 expression and histone modification. miR-20a was down-regulated in PMA/A23187-treated HMC-1 cells. miR-20a inhibited expression of pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β) and Interferon gamma (IFN-γ) while promoted Interleukin 10 (IL-10) production. miR-20a targeted HDAC4 and suppressed its expression, which contributed to epigenetically regulation of IL-10 expression by miR-20a. hsa-miR-20a-5p attenuates allergic inflammation in HMC-1 cells by targeting HDAC4.

Topics: Base Sequence; Calcimycin; Cell Line; Cytokines; Down-Regulation; Epigenesis, Genetic; Histone Deacetylases; Humans; Hypersensitivity; Inflammation; Inflammation Mediators; MicroRNAs; Ovalbumin; Repressor Proteins; Tetradecanoylphorbol Acetate

The activation of peroxisome proliferator-activated receptor γ (PPAR-γ) has been shown to attenuate allergic airway inflammation (AAI). To gain better understanding of mechanisms underlying this effect, the impact of rosiglitazone (RSG), a PPAR-γ agonist, on CD4

Topics: Animals; Asthma; CD4 Lymphocyte Count; Disease Models, Animal; Forkhead Transcription Factors; Gene Expression; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; Rosiglitazone; T-Lymphocytes, Regulatory; Treatment Outcome

Caffeic amides are derivatives of caffeic acid, which have antioxidant and anti-inflammatory properties, and high in vivo stability. The therapeutic effect of caffeic amides on allergic diseases, and especially on the maturation of bone marrow-derived dendritic cells (BM-DCs), remains unclear. In this study, we investigated the therapeutic potential of caffeic amides on allergic diseases by evaluating the maturation of DCs and evaluated their potential in inducing the differentiation of T. BM-DCs isolated from BALB/c mice were treated with different caffeic amide derivatives for 48 h and the expression of surface markers was analyzed by flow cytometry. The differentiation of CD4. Our results showed that among the six caffeic amides tested herein, only 36 M significantly inhibited the antigen-induced maturation of DCs associated with the expression of CD80, CD86, and major histocompatibility complex II (VC ovalbumin (OVA)+ thymic stromal lymphopoietin (TSLP) vs. 36 M OVA + TSLP). Additionally, the isolation and co-culture of antigen-specific CD4. Among the six caffeic amides tested herein, 36 M (N-octyl caffeamide) might possess therapeutic potential for allergic diseases.

Topics: Allergens; Amides; Animals; Anti-Allergic Agents; Bone Marrow Cells; Caffeic Acids; Cell Differentiation; Cytokines; Dendritic Cells; Disease Models, Animal; Female; Flow Cytometry; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Thymic Stromal Lymphopoietin

Despite a broad cell-type tropism, cytomegalovirus (CMV) is an evidentially pulmonary pathogen. Predilection for the lungs is of medical relevance in immunocompromised recipients of hematopoietic cell transplantation, in whom interstitial CMV pneumonia is a frequent and, if left untreated, fatal clinical manifestation of human CMV infection. A conceivable contribution of CMV to airway diseases of other etiology is an issue that so far attracted little medical attention. As the route of primary CMV infection upon host-to-host transmission in early childhood involves airway mucosa, coincidence of CMV airway infection and exposure to airborne environmental antigens is almost unavoidable. For investigating possible consequences of such a coincidence, we established a mouse model of airway co-exposure to CMV and ovalbumin (OVA) representing a protein antigen of an inherently low allergenic potential. Accordingly, intratracheal OVA exposure alone failed to sensitize for allergic airway disease (AAD) upon OVA aerosol challenge. In contrast, airway infection at the time of OVA sensitization predisposed for AAD that was characterized by airway inflammation, IgE secretion, thickening of airway epithelia, and goblet cell hyperplasia. This AAD histopathology was associated with a T helper type 2 (Th2) transcription profile in the lungs, including IL-4, IL-5, IL-9, and IL-25, known inducers of Th2-driven AAD. These symptoms were all prevented by a pre-challenge depletion of CD4+ T cells, but not of CD8+ T cells. As to the underlying mechanism, murine CMV activated migratory CD11b+ as well as CD103+ conventional dendritic cells (cDCs), which have been associated with Th2 cytokine-driven AAD and with antigen cross-presentation, respectively. This resulted in an enhanced OVA uptake and recruitment of the OVA-laden cDCs selectively to the draining tracheal lymph nodes for antigen presentation. We thus propose that CMV, through activation of migratory cDCs in the airway mucosa, can enhance the allergenic potential of otherwise poorly allergenic environmental protein antigens.

Topics: Allergens; Animals; Antigen Presentation; CD11 Antigens; Cytomegalovirus; Dendritic Cells; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Lung; Lung Diseases; Mice; Mice, Inbred C57BL; Ovalbumin; Th2 Cells; Virus Activation

Asthma is a chronic disease involving inflamed airways, which were previously demonstrated, can be modulated by the mesenchymal stem cells derived from induced pluripotent stem cells (iPSC-MSCs). However, the long-term effects of iPSC-MSCs in inflamed airways are still unidentified. This study investigated the long-term effects and potential mechanisms involved in the immunomodulatory effects of iPSC-MSCs in the chronic mouse asthma model.. Both human iPSC-MSCs and bone marrow (BM)-MSCs were transplanted into the long-term ovalbumin-induced mice before sensitization phase or during the challenge phase. Airway hyper-respnsiveness measurement, immunohistochemistry and ELISA were employed to assess the effects of MSCs. In addition, Smad2/3 levels were assessed by western blot analysis to investigate the possible mechanism involved.. The systemic administration of human iPSC-MSCs before the challenge protected the mice from the characters of the chronic allergic airway inflammation, in particular improving the airway remodeling and preventing fibrosis. In addition, the TGF-β1/Smad pathway was identified involved in the immunomodulatory effects of iPSC-MSCs on chronic allergic airway inflammation.. The study demonstrated that iPSC-MSCs are capable of preventing chronic allergic airway inflammation over a prolonged period, which further proved the iPSC-MSC therapeutic potential for allergic airway inflammation in a clinical scenario.

Topics: Animals; Bronchoalveolar Lavage Fluid; Chronic Disease; Cytokines; Disease Models, Animal; Female; Humans; Hypersensitivity; Induced Pluripotent Stem Cells; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Signal Transduction; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1

Spatial and temporal control of gene expression using cre/loxP technology is a major methodological advance for biomedical research. The ability to alter gene expression after an in vivo disease model has been established and allows researchers the opportunity to examine the impact of that gene on the perpetuation of a disease, a mechanistic insight that is arguably more therapeutically relevant than developmental mechanisms.We used the cre/LoxP technology in mice to show that β-arrestin-2, a gene previously shown to be required for the development of the asthma phenotype, is also required for the perpetuation of, at least, the airway hyperresponsiveness characteristic of that phenotype. Here we describe stepwise methods for the activation of the cre-loxP technology and induction of murine allergic inflammatory airway disease. We comment on the unanticipated problems encountered, which we speculate were a result of interactions between the allergen and β-arrestin-2 gene (Arrb2) regulation and the effect of tamoxifen on the asthma phenotype. The issues encountered here may be generally applicable to cre/loxP utilization in inflammatory disease models, especially if the gene of interest is associated with the inflammatory cascade.

Topics: Animals; Asthma; beta-Arrestin 2; Disease Models, Animal; Hypersensitivity; Inflammation; Mice, Knockout; Molecular Biology; Ovalbumin

We previously developed a test for detecting naturally occurring protein-induced skin sensitization based on the markers and criteria of the human cell-line activation test (h-CLAT) and showed that the h-CLAT was useful for assessing the allergenic potency of proteins. However, test proteins were contaminated with varying amounts of lipopolysaccharide (LPS), which might have contributed to the stimulation of CD86 and CD54 expression. In this study, we developed a method to exclude the effects of LPS in the assessment of skin sensitization by naturally occurring proteins. We tested two inhibitors [the caspase-1 inhibitor acetyl-Tyr-Val-Ala-Asp-chloromethylketone (Ac-YVAD-cmk; hereafter referred to as YVAD), which can mitigate the LPS-induced increases in CD54 expression, and polymyxin B (PMB), which suppresses the effect of LPS by binding to its lipid moiety (i.e., the toxic component of LPS)]. After a 24 hr exposure, YVAD and PMB reduced LPS-induced CD86 and CD54 expression. In particular, the effect of PMB was dependent upon pre-incubation time and temperature, with the most potent effect observed following pre-incubation at 37°C for 24 hr. Moreover, only pre-incubation with cell-culture medium (CCM) at 37°C for 24 hr showed an inhibitory effect similar to that of PMB, with this result possibly caused by components of CCM binding to LPS. Similar effects were observed in the presence of ovalbumin (with 1070 EU/mg LPS) and ovomucoid, and lysozyme (with 2.82 and 0.234 EU/mg LPS, respectively) in CCM. These results indicated that PMB and CCM effectively eliminated the effects of LPS during assessment of protein allergenicity, thereby allowing a more accurate evaluation of the potential of proteins to induce skin sensitization.

Topics: Amino Acid Chloromethyl Ketones; B7-2 Antigen; Culture Media; Gene Expression; Humans; Hypersensitivity; Immunization; Intercellular Adhesion Molecule-1; Lipopolysaccharides; Muramidase; Ovalbumin; Ovomucin; Polymyxin B; Protein Binding; Proteins; Skin; Skin Tests; Temperature; THP-1 Cells; Time Factors

Topics: Animals; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Inflammation; Intracellular Signaling Peptides and Proteins; Lipopolysaccharides; Mice; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; RAW 264.7 Cells; Teichoic Acids; Toll-Like Receptor 2

Ziziphora clinopodioides has been frequently used as an anti asthmatic plant in traditional medication. Recent work explores the anti-asthmatic activity of Z. clinopodioides in allergen-induced asthmatic mice. Intraperitoneal sensitization followed by intranasal challenge were given with ovalbumin (allergen) to develop allergic asthma. Investigational groups of animals were administered with drug methylprednisolone (MP) (15 mg/kg body weight), n-hexane fraction, ethylacetate fraction, and methanolic extract of Z. clinopodioides extract (500 mg/kg b.w.) for successive 07 days. Hematoxyline and eosin (H&E) and periodic acid-Schiff (PAS) stains were used to evaluate histopathological parameters on lung tissues. As an index of lungs tissues edema, wet/dry weight ratio of lungs was determined. Evaluation of expression levels of AQP1, AQP5, IL4, and IL5 was conducted by using RT-PCR. The data exhibited that both Z. clinopodioides and MP attenuated differential and total leukocyte counts in hematological examination i.e. in BALF and blood. Treatment with Z. clinopodioides also caused suppression of inflammatory cell infiltration and expression levels of IL4 and IL5, the later could have caused attenuation of pulmonary inflammation. The study also found decline in lung wet/dry ratio and goblet cellh hyperplasia in treated groups which indicates amelioration of lung edema. Treatment with Z. clinopodioides significantly increased the expression levels of aquaporin-1 and -5, which could have led to reduction in lung edema. The treatment with MP showed comparable results to Z. clinopodioides. Current investigation revealed that Z. clinopodioides possessed anti-asthmatic property which might be accredited to upregulagted AQP1 and AQP5 levels and downregulated IL4 and IL5 levels.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Aquaporins; Asthma; Cytokines; Disease Models, Animal; Down-Regulation; Female; Hypersensitivity; Inflammation; Mentha; Methylprednisolone; Mice; Ovalbumin; Plant Extracts; Pulmonary Edema; Up-Regulation

To investigate the effects of exposure to diethyl phthalate(DEHP) during pregnancy and lactation in respiratory allergy of offspring Wistar rats.. To establish the maternal DEHP exposure model: 36 healthy 2-month-old female Wistar rats were divided into three different dose groups. From GD0, each group of pregnant mice were given different concentrations of DEHP(0, 30, 300 mg/(kg·d)) until to the newborn weaning(PND21). After birth, one of offspring was selected from each cage in different dose groups. At PND21 and PND28 the offspring were sensitized by intraperitoneal injection(i. p. ) OVA and continuous nasal sensitization at PND32, PND33, PND34. Bronchoalveolar lavage fluid(BALF) and lung tissue were collected at PND35 with non-sensitized groups. ELISA detected the secretion of Th2 cytokine interleukin(IL)-4, IL-5 and IL-13 in BALF, and the pathological changes of allergic inflammation in lung tissues were observed by HE and PAS staining. Expression of epithelium-derived factor IL-33 detected by immunohistochemistry.. In BALF, compared with the control group, the total number of inflammatory cells and eosinophils in the DEHP0+OVA group increased to(131. 500±25. 548)×10~4, (32. 000±10. 079)×10~4(P<0. 05); The total number of inflammatory cells and eosinophils in the DEHP30+OVA group increased to(156. 167±17. 994)×10~4, (16. 331±6. 667)×10~4(P<0. 05); and the total number of inflammatory cells and eosinophils in the DEHP300+OVA group increased to(172. 167±19. 994)×10~4, (55. 000±17. 018)×10~4(P<0. 05). After adding different doses of DEHP and OVA, compared with the control group, The expression levels of IL-4, IL-5, IL-13 in DEHP0+OVA group increased to(38. 401±6. 594) pg/mL(P>0. 05), (30. 026±2. 756) pg/mL(P<0. 05), (13. 806±4. 355) pg/mL(P<0. 05); The expression levels of IL-4, IL-5 and IL-13 in DEHP30+OVA group increased to(57. 733±7. 293) pg/mL(P<0. 05) and(31. 544±1. 043) pg/mL(P>0. 05), (18. 068±1. 497) pg/mL(P<0. 05); The expression levels of IL-4, IL-5 and IL-13 in DEHP300+OVA group increased to(54. 943±6. 049) pg/mL(P>0. 05) and(32. 377±3. 739)pg/mL(P>0. 05), (20. 168±0. 939) pg/mL(P<0. 05), respectively. The tissue section of all the DEHP+saline groups could be observed that no obvious allergic inflammatory reaction, meanwhile all the DEHP+OVA groups had a relatively obvious allergic reaction and were severe with the increase of DEHP dose. Immunohistochemistry showed no significant increase in the expression of IL-33 in the DEHP+saline groups, while the expression of IL-33 in the DEHP+OVA groups increased with a certain dose response.. Exposure to DEHP during pregnancy and lactation will aggravate the sensitization reaction of offspring, the possible mechanism that DEHP increase the Th2 type of immune response may be the overexpression of IL-33 in the epithelial cells.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Humans; Hypersensitivity; Interleukin-33; Lactation; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalic Acids; Pregnancy; Rats; Rats, Wistar

Corticosteroids commonly prescribed in asthma show several side-effects. Relatively non-toxic andrographolide (AG) has an anti-asthmatic potential. But its poor bioavailability and short plasma half-life constrain its efficacy. To overcome them, we encapsulated AG in nanoparticle (AGNP) and evaluated AGNP for anti-asthmatic efficacy on murine asthma model by oral/pulmonary delivery. AGNP had 5.47% drug loading with a sustained drug release in vitro. Plasma and lung pharmacokinetic data showed predominantly improved AG-bioavailability upon AGNP administered orally/by pulmonary route. Cell numbers, IL-4, IL-5, and IL-13 levels in broncho-alveolar lavage fluid and serum IgE content were reduced significantly after administration of AGNP compared to free-AG treatment. AGNP-mediated suppression of NF-κβ was predominantly more compared to free-AG. Further, pulmonary route showed better therapeutic performance. In conclusion, AGNP effectively controlled mild and severe asthma and the pulmonary administration of AGNP was more efficacious than the oral route.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Diterpenes; Drug Liberation; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Male; Mice, Inbred C57BL; Nanoparticles; NF-kappa B; Ovalbumin; Particle Size; Rats, Sprague-Dawley; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Tissue Distribution

Topics: Animals; Antigens, Plant; B-Lymphocytes; B7-2 Antigen; CD40 Antigens; Dendritic Cells; Gene Silencing; Hypersensitivity; Immunization; Immunoglobulin E; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Plant Proteins; Rhinitis, Allergic

Helminths immunomodulate the host immune system by secreting proteins to create an inhibitory environment as a strategy for survival in the host. As a bystander effect, this balances the host immune system to reduce hypersensitivity to allergens or autoantigens. Based on this, helminth therapy has been used to treat some allergic or autoimmune diseases. As a tissue-dwelling helminth, Trichinella spiralis infection has been identified to have strong immunomodulatory effects; the effective components in the worm have not yet been identified.. The soluble extracts of T. spiralis adult worms and muscle larvae were used to treat airway inflammation before and after an ovalbumin (OVA)-sensitization/challenge in an OVA-induced asthma mouse model. The therapeutic effects were observed by measuring the level of inflammation in the lungs.. The soluble products derived from T. spiralis parasites, especially from adult worms, were able to ameliorate OVA-induced airway inflammatory responses which were associated with reduced eosinophil infiltration, OVA-specific IgE, Th2 cytokine IL-4, and increased IL-10 and TGF-β. The stimulation of the Treg response may contribute to the alleviated allergic inflammation.. Trichinella spiralis worm extracts stimulate regulatory cytokines that are associated with reduced allergic airway inflammation. The identification of effective components in the adult worm extracts will be a crucial approach for developing a novel therapeutic for allergic and autoimmune diseases.

Topics: Animals; Anti-Allergic Agents; Asthma; Cytokines; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells; Tissue Extracts; Trichinella spiralis

Linalool is a natural product present in fruits and aromatic plants with biological activities. Researchers have reported that the inhalation of linalool exerts anti-inflammatory activities. In this study, we examined the therapeutic effects of linalool on airway inflammation and mucus overproduction in mice with allergic asthma. Oral administration of linalool significantly inhibited the levels of eosinophil numbers, Th2 cytokines and immunoglobulin E (IgE) caused by ovalbumin (OVA) exposure. Linalool exerted preventive effects against the influx of inflammatory cells and mucus hypersecretion in the lung tissues. Linalool also dose-dependently decreased the levels of inducible nitric oxide synthase (iNOS) expression and protein kinase B (AKT) activation in the lung tissues. Linalool effectively downregulated the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) caused by OVA exposure. Furthermore, linalool exerted inhibitory effect on OVA-induced airway hyperresponsiveness (AHR). In the in vitro study, the increased secretion of MCP-1 was attenuated with linalool treatment in lipopolysaccharide (LPS)-stimulated H292 airway epithelial cells. In conclusion, linalool effectively exerts a protective role in OVA-induced airway inflammation and mucus hypersecretion, and its protective effects are closely related to the downregulation of inflammatory mediators and MAPKs/NF-κB signaling.

Topics: Acyclic Monoterpenes; Administration, Oral; Allergens; Animals; Anti-Inflammatory Agents; Asthma; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Humans; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Nitric Oxide Synthase Type II; Ovalbumin; Respiratory Hypersensitivity; Respiratory Mucosa; Th2 Cells

Perinatal exposures are associated with altered risks of childhood allergy. Human studies and our previous work suggest that restricted growth in utero (IUGR) is protective against allergic disease. The mechanisms are not clearly defined, but reduced fetal abundance and altered metabolism of methyl donors are hypothesized as possible underlying mechanisms. Therefore, we examined whether late-gestation maternal dietary methyl donor and cofactor supplementation of the placentally restricted (PR) sheep pregnancy would reverse allergic protection in progeny. Allergic outcomes were compared between progeny from control pregnancies (CON; n = 49), from PR pregnancies without intervention (PR; n = 28), and from PR pregnancies where the dam was fed a methyl donor plus cofactor supplement from day 120 of pregnancy until delivery (PR + Methyl; n = 25). Both PR and PR + Methyl progeny were smaller than CON; supplementation did not alter birth size. PR was protective against cutaneous hypersensitivity responses to ovalbumin (OVA; P < 0.01 in singletons). Cutaneous hypersensitivity responses to OVA in PR + Methyl progeny were intermediate to and not different from the responses of CON and PR sheep. Cutaneous hypersensitivity responses to house dust mites did not differ between treatments. In singleton progeny, upper dermal mast cell density was greater in PR + Methyl than in PR or CON (each P < 0.05). The differences in the cutaneous allergic response were not explained by treatment effects on circulating immune cells or antibodies. Our results suggest that mechanisms underlying in utero programming of allergic susceptibility by IUGR and methyl donor availability may differ and imply that late-gestation methyl donor supplementation may increase allergy risk.

Topics: Animals; Cobalt; Dermatitis; Dietary Supplements; Disease Models, Animal; DNA Methylation; Female; Fetal Growth Retardation; Folic Acid; Gestational Age; Hypersensitivity; Immunoglobulin E; Mast Cells; Methionine; Ovalbumin; Placenta; Pregnancy; Prenatal Exposure Delayed Effects; Pyroglyphidae; Sheep, Domestic; Skin; Sulfur

Topics: Adoptive Transfer; Animals; Asthma; Calcium-Binding Proteins; Cell Differentiation; Cell Proliferation; Dendritic Cells; Down-Regulation; Female; Hypersensitivity; Immunoglobulin E; Intercellular Signaling Peptides and Proteins; Interleukin-12; Lymphocyte Activation; Mice; Ovalbumin; Th1 Cells; Th2 Cells

T lymphocytes express not only cell membrane ORAI calcium release-activated calcium modulator 1 but also voltage-gated calcium channel (Ca. We investigated the role of β and α2δ auxiliary subunits on Ca. We used Ca. Mouse and human T. These results stress the role of Ca

Topics: Acute Disease; Allergens; Animals; Calcium Channels, L-Type; Female; Hypersensitivity; Inflammation; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Protein Subunits; T-Lymphocytes

Respiratory IgE-sensitization to innocuous antigens increases the risk for developing diseases such as allergic asthma. Dendritic cells (DC) residing in the airways orchestrate the immune response following antigen exposure and their ability to sample and present antigens to naïve T cells in airway draining lymph nodes contributes to allergen-specific IgE-sensitization. In order to characterize inhaled antigen capture and presentation by DC subtypes in vivo, we used an adjuvant-free respiratory sensitization model using two genetically distinct rat strains, one of which is naturally resistant and the other naturally susceptible to allergic sensitization. Upon multiple exposures to ovalbumin (OVA), the susceptible strain developed OVA-specific IgE and airway inflammation, whereas the resistant strain did not. Using fluorescently tagged OVA and flow cytometry, we demonstrated significant differences in antigen uptake efficiency and presentation associated with either IgE-sensitization or resistance to allergen exposures in respective strains. We further identified CD4

Topics: Animals; Dendritic Cells; Hypersensitivity; Immunization; Immunoglobulin E; Inflammation; Lung; Lymph Nodes; Ovalbumin; Phenotype; Rats, Inbred BN; T-Lymphocytes, Regulatory

Asthma and chronic obstructive pulmonary disease (COPD), chronic airway inflammatory diseases characterized by airflow limitation, have different etiologies and pathophysiologies. Asthma-COPD Overlap (ACO) has recently been used for patients with mixed asthma and COPD. The pathophysiological mechanisms of ACO have not been clearly understood due to the lack of an appropriate murine model. To investigate its pathophysiology, we examined a murine model by allergen challenge in surfactant protein-D (SP-D)-deficient mice that spontaneously developed pulmonary emphysema. SP-D-deficient mice were sensitized and challenged by ovalbumin (OVA). Lungs and bronchoalveolar lavage fluid (BALF) were collected for analysis, and static lung compliance and airway hyperresponsiveness (AHR) were measured 48 h after the last OVA challenge. In SP-D-deficient, naïve, or OVA-challenged mice, the mean linear intercept and static lung compliance were increased compared with wild-type (WT) mice. There was no significant difference in goblet cell hyperplasia and the gene expression of Mucin 5AC (MUC5AC) between SP-D-deficient and WT OVA-challenged mice. In SP-D-deficient OVA-challenged mice, airway hyperresponsiveness was significantly enhanced despite the lower eosinophil count and the concentration of interleukin (IL)-5 and IL-13 in BALF compared with WT OVA-challenged mice at 120 ventilations per minute. When mice were ventilated at a lower ventilation frequency of 100 ventilations per minute, elevated airway hyperresponsiveness in SP-D-deficient OVA-challenged mice was diminished. This model of emphysematous change with allergic airway inflammation raises the possibility that frequency-dependent airway hyperresponsiveness may be involved in the pathophysiology of ACO.

Topics: Animals; Asthma; Disease Models, Animal; Hypersensitivity; Inflammation; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Pulmonary Emphysema; Pulmonary Surfactant-Associated Protein D; Respiratory Hypersensitivity

Nanoparticle (NP)-based vaccines are attractive immunotherapy tools because of their capability to codeliver antigen and adjuvant to antigen-presenting cells. Their cellular distribution and serum protein interaction ("protein corona") after systemic administration and their effect on the functional properties of NPs is poorly understood.. We analyzed the relevance of the protein corona on cell type-selective uptake of dextran-coated NPs and determined the outcome of vaccination with NPs that codeliver antigen and adjuvant in disease models of allergy.. The role of protein corona constituents for cellular binding/uptake of dextran-coated ferrous nanoparticles (DEX-NPs) was analyzed both in vitro and in vivo. DEX-NPs conjugated with the model antigen ovalbumin (OVA) and immunostimulatory CpG-rich oligodeoxynucleotides were administered to monitor the induction of cellular and humoral immune responses. Therapeutic effects of this DEX-NP vaccine in mouse models of OVA-induced anaphylaxis and allergic asthma were assessed.. DEX-NPs triggered lectin-induced complement activation, yielding deposition of activated complement factor 3 on the DEX-NP surface. In the spleen DEX-NPs targeted predominantly B cells through complement receptors 1 and 2. The DEX-NP vaccine elicited much stronger OVA-specific IgG. Opsonization of lectin-coated NPs by activated complement components results in selective B-cell targeting. The intrinsic B-cell targeting property of lectin-coated NPs can be exploited for treatment of allergic immune responses.

Topics: Anaphylaxis; Animals; Antigens; B-Lymphocytes; Dextrans; Drug Carriers; Female; Ferrous Compounds; Hypersensitivity; Lectins; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Nanoparticles; Oligodeoxyribonucleotides; Ovalbumin; Protein Corona; T-Lymphocytes; Vaccines

Asthma is a chronic disease affecting up to 10% of the Australian population for which medical treatment is solely aimed at relief of symptoms rather than prevention of disease. Evidence from animal and human studies demonstrates a strong link between viral respiratory infections, atopy and the development of asthma. Type I IFNs include IFNα and IFNβ, with subtype expression tailored toward the specific viral infection. We hypothesized that exposure to type I IFNs and allergen may interfere with the healthy response to innocuous airway antigen exposure. In this study, we use an ovalbumin (OVA)-induced BALB/c model of experimental allergic airways disease, where pre-exposure of the airways to OVA is protective against allergen sensitization, leading to allergen tolerance. We investigated airways pre-exposure with OVA and type I IFNs on development of allergic airways disease. We demonstrate restoration of allergic airways disease on pre-exposure with allergen and IFNβ, and not IFNα. Dysfunction in tolerance led to changes in dendritic cell antigen capture/traffic, T-cell and B-cell responses. Furthermore, exposure to IFNβ with ongoing allergen exposure led to the development of hallmark asthma features, including OVA-specific IgE and airways eosinophilia. Data indicate a role for IFNβ in linking viral infection and allergy.

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Eosinophils; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Interferon-alpha; Interferon-beta; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Transcriptional repressor B-cell lymphoma 6 (Bcl6) appears to regulate T

Topics: Animals; Antigens; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Hypersensitivity; Mice, Transgenic; Ovalbumin; Phenotype; Proto-Oncogene Proteins c-bcl-6

We previously reported that the biologically active form of histamine releasing factor (HRF) is dimerized translationally controlled tumor protein (dTCTP) which is involved in a number of allergic diseases.. Hoping that agents that modulate dTCTP may provide new therapeutic targets to allergic inflammatory diseases, we screened a library of natural products for substances that inhibit dTCTP. One such inhibitor we found was dehydrocostus lactone (DCL), a natural sesquiterpene present in rhizome of Saussurea lappa Clarke, the subject of this study.. We evaluated the therapeutic efficacy of DCL in a mouse model of ovalbumin (OVA)-induced allergic airway inflammation, employing the ELISA system using BEAS-2B cells and splenocytes, and confirmed that DCL interacts with dTCTP using SPR assay.. DCL inhibited dTCTP-induced secretion of IL-8 in BEAS-2B cells. From kinetic analysis of dTCTP and DCL, we found that K. DCL's therapeutic potential in allergic airway inflammation is based on its anti-inflammatory activity of suppressing the function of dTCTP.

Topics: Animals; Asthma; Biomarkers, Tumor; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Kinetics; Lactones; Mice, Inbred BALB C; Ovalbumin; Protein Multimerization; Saussurea; Sesquiterpenes; Surface Plasmon Resonance; Tumor Protein, Translationally-Controlled 1

Exosomes are nanovesicles originating from multivesicular bodies that have complex functions and significant therapeutic effects in many diseases. In the present study, we successfully extracted exosomes from Pseudomonas aeruginosa and assessed the effect of those exosomes on the development of the allergic response in two types of classic asthma models.. Female BALB/c mice were administrated with P. aeruginosa-derived exosomes 1 week before ovalbumin (OVA) or house dust mite (HDM) sensitization. Bronchoalveolar lavage fluid, serums and lung tissues were collected and analyzed for pathophysiology and immune responses.. Our results demonstrated that P. aeruginosa-derived exosomes inhibited the development of airway hyper-responsiveness (AHR), peribronchial and perivascular inflammation in lung tissues and the level of serum IgE. Moreover, this protective effect was associated with an increase in the regulatory T cell (T. In conclusion, these observations demonstrated that P. aeruginosa-derived exosomes could induce protection against allergic sensitization in asthma mice, and our study provided a new insight to prevent allergic diseases.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Exosomes; Female; Hypersensitivity; Immune System; Immunoglobulin E; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pseudomonas aeruginosa; T-Lymphocytes, Regulatory

This study aimed to demonstrate whether exposure to bifidobacteria during early life influences immunity and alleviates the risk of immunoglobulin E (IgE)-mediated allergies in adulthood. BALB/c neonatal mice (n=54) were administered with a lyophilised cell preparation of Bifidobacterium bifidum TMC3115 (TMC3115) for 3 weeks. Following the intervention, the mice were immunised with intraperitoneal ovalbumin (OVA). The morphology and function of the intestinal epithelium were determined using histopathological examinations. Intestinal microbiota was detected using quantitative PCR and characterised using next-generation sequencing of 16S rRNA genes from faecal DNA. Caecal short-chain fatty acids (SCFAs) were measured using gas chromatography-mass spectrometry. Serum levels of tumour necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, and immunoglobulin E (IgE) and the percentage of splenic CD4+ T cells were examined using enzyme-linked immunosorbent assay and flow cytometry, respectively. TMC3115 did not significantly affect body weight, and cause any severe systemic inflammation or other clinical symptoms among the neonatal or adult mice, although the crypt depths and Muc2-positive cells in some intestinal segments of neonatal mice were significantly lower than control. Oral TMC3115 administration significantly increased faecal microbial diversity, relative abundance of Bacteroidetes and caecal SCFAs production in neonatal mice. Following the intervention, neonatal mice treated with TMC3115 exhibited less increase in serum IgE levels induced by OVA in adults and significantly higher TNF-α and IL-10 levels than in control. Our findings indicate that the oral administration of bifidobacteria, particularly certain strains, such as TMC3115, during early life could alleviate the risk of IgE-mediated allergies in adult host animals. Modifications of intestinal microbiota, SCFAs metabolism and anti-inflammatory cytokine IL-10 production by bifidobacteria may at least in part be a key mechanism underlying the effect of bifidobacteria on the IgE-mediated immune sensitivity of hosts to attacks by allergens at both neonatal and adult stages.

Topics: Administration, Oral; Animals; Animals, Newborn; Bifidobacterium bifidum; Enzyme-Linked Immunosorbent Assay; Female; Gastrointestinal Microbiome; Humans; Hypersensitivity; Immunoglobulin E; Infant, Newborn; Infant, Newborn, Diseases; Interleukin-10; Interleukin-6; Intestines; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Tumor Necrosis Factor-alpha

The mechanisms of allergen immunotherapy are not fully elucidated. Here, we sought to develop a murine model to demonstrate the effectiveness of subcutaneous immunotherapy (SCIT) for allergic responses. As excessive antigen dosages may induce immune tolerance in sensitized mice, the effects of SCIT were assessed by varying the antigen dosage. The mechanisms of SCIT were analyzed by focusing on the induction of Foxp3. The maximum effects of SCIT were observed with 1 mg/animal of OVA for airway inflammation induced by 5 μg/animal of OVA, in which airway eosinophilia and Th2 cytokine production were markedly suppressed. The increase in the OVA-specific IgE level was significantly suppressed by SCIT. The development of bronchial epithelial thickening and mucus accumulation were also suppressed by SCIT. Concomitantly, IL-10-producing Foxp3. We successfully developed an airway allergic model for SCIT. It was suggested that most of IL-10-producing Foxp3

Topics: Allergens; Animals; Asthma; Cells, Cultured; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Eosinophils; Forkhead Transcription Factors; Humans; Hypersensitivity; Immune Tolerance; Infusions, Subcutaneous; Interleukin-10; Lung; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; T-Lymphocytes, Regulatory; Th1-Th2 Balance

The importance of developing new animal models to assess the pathogenesis of glucocorticoid (GC)-insensitive asthma has been stressed. Because of the asthma-prone background of A/J mice, we hypothesized that asthma changes in these animals would be or become resistant to GCs under repeated exposures to an allergen. A/J mice were challenged with OVA for 2 or 4 consecutive d, starting on day 19 postsensitization. Oral dexamethasone or inhaled budesonide were given 1 h before challenge, and analyses were done 24 h after the last challenge. Airway hyperreactivity, leukocyte infiltration, tissue remodeling, and cytokine levels as well as phosphorylated GC receptor (p-GCR), p-GATA-3, p-p38, MAPK phosphatase-1 (MKP-1), and GC-induced leucine zipper (GILZ) levels were assessed. A/J mice subjected to two daily consecutive challenges reacted with airway hyperreactivity, subepithelial fibrosis, and marked accumulation of eosinophils in both bronchoalveolar lavage fluid and peribronchial space, all of which were clearly sensitive to dexamethasone and budesonide. Conversely, under four provocations, most of these changes were steroid resistant. A significant reduction in p-GCR/GCR ratio following 4- but not 2-d treatment was observed, as compared with untreated positive control. Accordingly, steroid efficacy to transactivate MKP-1 and GILZ and to downregulate p-p38, p-GATA-3 as well as proinflammatory cytokine levels was also seen after two but not four provocations. In conclusion, we report that repeated allergen exposure causes GC-insensitive asthma in A/J mice in a mechanism associated with decrease in GCR availability and subsequent loss of steroid capacity to modulate pivotal regulatory proteins, such as GATA-3, p-p38, MKP-1, and GILZ.

Topics: Allergens; Animals; Asthma; Biological Availability; Bronchoalveolar Lavage Fluid; Budesonide; Cytokines; Dexamethasone; Disease Models, Animal; Down-Regulation; Eosinophils; Glucocorticoids; Hypersensitivity; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Glucocorticoid; Steroids; Transcriptional Activation

Topics: Acridines; Allergens; Animals; B-Lymphocytes; Cells, Cultured; Female; G-Quadruplexes; Hypersensitivity; Immunoglobulin Class Switching; Inflammation; Mice, Inbred BALB C; Ovalbumin

Objective To investigate the changes of 5-HT (serotonin) signaling system in allergic diarrhea mice sensitized with ovalbumin (OVA). Methods The seven-to-eight-week-old BALB/c female mice were randomly divided into model group, sodium chromate group and negative control group. The model group and sodium chromate group were intraperitoneally injected with OVAI (50 μg per mouse) at day 0 and day 14 respectively. And starting from the 28th day, OVAII was orally administered (50 mg per mousee) every other day (8 times in total), and the sodium chromate group was given the sodium chromate (78.0 mg/kg) before the oral administration of OVA every other day (8 times in total). The allergic symptoms, including the systemic score, faeces score and body temperature were recorded following the OVA administration for sensitization. The mice were executed 43 days later. Eyeball blood sample was collected, and then serum was seperated by centrifugation, the gastric tissues was taken out. The serum OVA-specific IgE (OVA-SIgE) was detected by ELISA. The serum content of 5-HT and its related metabolites including kynurenine (KYN), tryptophan (TRP), 5-hydroxytryptophan (5-HTP), and 5-hydroxyindoleacetic acid (5-HIAA) were examined by liquid chromatography-mass spectrometry (LC-MS). The mRNA levels of tryptophan hydroxylase-1 (TPH1), indolamine-2, 3-dioxygenase 1 (IDO1), monoamine oxidase A (MAO-A), 5-hydroxytryptamine 1A receptor (HTR1A), 5-hydroxytryptamine 3 receptor (HTR3), 5-hydroxytryptamine 4 receptor (HTR4) and serotonin reuptake transporter (SERT) were determined by real-time quantitative PCR. Results OVA sensitization caused severe allergic diarrhea in mice. Serum OVA-SIgE increased significantly in mice sensitized by OVA. serum KYN increased remarkably, while 5-HT, 5-HIAA and 5-HTP decreased significantly. The mRNA levels of IDO1, HTR1A and HTR3A increased in gastric tissues, while the levels of TPH1 and MAO-A mRNA decreased. Compared with the model group, the sodium chromate group had lowed systemic score, faeces score, body temperature and OVA-SIgE as well as diarrhea rate. The mRNA levels of 5-HIAA and MAO-A increased in the gastric tissues, and IDO1, 5-HT1A and 5-HT3A mRNAs decreased in the sodium chromate group. Conclusion The serotonin signaling system in ovalbumin-sensitized allergic diarrhea mice has been activated. The administration of sodium chromate can alleviate the allergic symptoms, and change the levels of serum metabolites and the gene expressions

Topics: Animals; Colitis; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Serotonin

How respiratory syncytial virus (RSV) influences the development of ovalbumin (OVA)-induced asthma remains elusive. As potent T helper (Th)2 cytokine producers, group 2 innate lymphoid cells (ILC2s) are known to serve important functions in the pathogenesis of allergic inflammation. However, how RSV infection affects innate immunity, especially with regard to the function of ILC2s in OVA-induced allergic airway inflammation, is largely unknown.. RSV was used to infect adult BALB/c mice intranasally prior to sensitization and subsequent challenge with OVA. ILC2 frequencies and Th2 cytokine production by ILC2s were assessed by flow cytometry. Cytokine levels were detected both by real-time PCR and ELISA.. Previous infection with RSV attenuated airway inflammation and decreased Th2 cytokine production in mice sensitized and challenged with OVA. Furthermore, previous infection with RSV inhibited the influx of ILC2s into the lung, and constrained their Th2 cytokine production. Adoptive transfer of ILC2s increased asthma-associated airway inflammation in mice previously infected with RSV. These results indicate that previous infection with RSV prevents OVA-induced asthma development via inhibition of ILC2s. Previous infection with RSV attenuated IL-33 production in lung tissue and reduced relative ST2L expression in lung ILC2s, meaning that previous infection with RSV may alter ILC2 function via the IL-33/ST2 signaling pathway.. These results demonstrate that previous infection with RSV attenuates OVA-induced airway inflammation by inhibiting the recruitment and Th2 cytokine production of ILC2s via the IL-33/ST2 pathway.

Topics: Allergens; Animals; Cells, Cultured; Disease Models, Animal; Female; Humans; Hypersensitivity; Immunity, Innate; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Lung; Lymphocytes; Mice; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Signal Transduction; Th2 Cells

Great interest has been taken in the use of beneficial bacteria for allergic diseases recently, but the underlying mechanisms through which probiotics induces immune regulation or immune tolerance are poorly understood. We aimed to explore whether Lactobacillus rhamnosus GG (LGG)-induced beneficial effect relates to the change of microbiota. LGG was administered orally to mouse model of ovalbumin (OVA)-induced allergic airway inflammation. Our findings manifested that LGG-treatment contributes to protect against OVA-induced allergic airway inflammation by expanding mesenteric CD103

Topics: Animals; Disease Models, Animal; Gastrointestinal Microbiome; Hypersensitivity; Immune Tolerance; Immunologic Factors; Inflammation; Lacticaseibacillus rhamnosus; Lung; Male; Mice; Mice, Inbred BALB C; Microbiota; Ovalbumin; Probiotics; Protective Factors; T-Lymphocytes, Regulatory

Intestinal epithelial cells (IECs) are known to regulate allergic sensitization. We addressed the role of the intrinsic IKKβ signaling in IECs in the effector phase of allergy following oral allergen challenge and its impact on the severity of responses is poorly. Upon orally sensitization by co-administration of ovalbumin with cholera toxin as adjuvant, wild-type and mice lacking IKKβ in IECs (IKKβ

Topics: Administration, Oral; Allergens; Animals; Chemokine CCL11; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunoglobulin E; Intestinal Mucosa; Mice; Mice, Knockout; NF-kappa B; Ovalbumin; Severity of Illness Index

The efficacy of heparins and low-MW-heparins (LMWH) against human asthma has been known for decades. However, the clinical utility of these compounds has been hampered by their anticoagulant properties. Much effort has been put into harnessing the anti-inflammatory properties of LMWH but none have been used as therapy for asthma. Sulfated-non-anticoagulant heparin (S-NACH) is an ultra-LMWH with no systemic anticoagulant effects.. The present study explored the potential of S-NACH in blocking allergic asthma and examined the potential mechanism by which it exerts its effects.. Acute and chronic ovalbumin-based mouse models of asthma, splenocytes, and a lung epithelial cell line were used. Mice were challenged with aerosolized ovalbumin and administered S-NACH or saline 30 min after each ovalbumin challenge.. Sulfated-non-anticoagulant heparin administration in mice promoted a robust reduction in airway eosinophilia, mucus production, and airway hyperresponsiveness even after chronic repeated challenges with ovalbumin. Such effects were linked to suppression of Th2 cytokines IL-4/IL-5/IL-13/GM-CSF and ovalbumin-specific IgE without any effect on IFN-γ. S-NACH also reduced lung fibrosis in mice that were chronically-exposed to ovalbumin. These protective effects of S-NACH may be attributed to modulation of the IL-4/JAK1 signal transduction pathway through an inhibition of STAT6 phosphorylation and a subsequent inhibition of GATA-3 and inducible NO synthase expression. The effect of the drug on STAT6 phosphorylation coincided with a reduction in JAK1 phosphorylation upon IL-4 treatment. The protective effects of S-NACH treatment was associated with reduction of the basal expression of the two isoforms of arginase ARG1 and ARG2 in lung epithelial cells.. Our study demonstrates that S-NACH constitutes an opportunity to benefit from the well-known anti-asthma properties of heparins/LMWH while bypassing the risk of bleeding. Our results show, for the first time, that such anti-asthma effects may be associated with reduction of the IL-4/JAK1/STAT6 pathway.

Topics: A549 Cells; Animals; Anti-Inflammatory Agents; Anticoagulants; Asthma; Cell Line; Cell Separation; Disease Models, Animal; Flow Cytometry; Hemorrhage; Heparin; Humans; Hypersensitivity; Interleukin-4; Janus Kinase 1; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Signal Transduction; Spleen; STAT6 Transcription Factor; Th2 Cells

Most allergic reactions are induced by mast cell activation. Mast cells play vital roles in the pathogenesis of allergic diseases. Bisdemethoxycurcumin (BDMC), a natural curcuminoid, has potential anti-allergic effects. Hence, we explored the effect of BDMC on mast cell-mediated allergic diseases. The study proved that BDMC suppresses β-hexosaminidase release, granule release, and membrane ruffling in monoclonal anti-2,4,6-dinitrophenyl-immunoglobulin (Ig) E/human serum albumin (DNP-IgE/HSA)-stimulated rat basophilic leukaemia cells (RBL-2H3 cells), and BDMC suppressed ovalbumin (OVA)-induced allergic rhinitis (AR) symptoms and OVA-specific IgE levels in AR mice. Furthermore, BDMC increased the survival of compound 48/80 anaphylaxis shock mice and elevated the decreased rectal temperature in OVA-induced active systemic anaphylaxis mice. These findings indicate that BDMC regulates the degranulation of mast cells, demonstrating its potential in the treatment of mast cell-induced allergic reactions.

Topics: Anaphylaxis; Animals; Cell Line; Curcumin; Diarylheptanoids; Hypersensitivity; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; p-Methoxy-N-methylphenethylamine; Rats; Rhinitis, Allergic

Epidemiologic studies indicate that exposure to psychosocial stress in early childhood is a risk factor of adult-onset asthma, but the mechanisms of this relationship are poorly understood. Therefore, we examined whether early-life stress increases susceptibility to adult-onset asthma by inhibiting the development of respiratory tolerance. Neonatal BALB/c female mice were aerosolized with ovalbumin (OVA) to induce immune tolerance prior to immune sensitization with an intraperitoneal injection of OVA and the adjuvant aluminum hydroxide. Maternal separation (MS) was applied as an early-life stressor during the induction phase of immune tolerance. The mice were challenged with OVA aerosol in adulthood, and allergic airway responses were evaluated, including airway hyper-responsiveness to inhaled methacholine, inflammatory cell infiltration, bronchoalveolar lavage fluid levels of interleukin (IL)-4, IL-5, and IL-13, and serum OVA-specific IgE. We then evaluated the effects of MS on the development of regulatory T (Treg) cells in bronchial lymph nodes (BLN) and on splenocyte proliferation and cytokine expression. In mice that underwent MS and OVA tolerization, the allergic airway responses and OVA-induced proliferation and IL-4 expression of splenocytes were significantly enhanced. Furthermore, exposure to MS was associated with a lower number of Treg cells in the BLN. These findings suggest that exposure to early-life stress prevents the acquisition of respiratory tolerance to inhaled antigen due to insufficient Treg cell development, resulting in Th2-biased sensitization and asthma onset. We provide the evidence for inhibitory effects of early-life stress on immune tolerance. The present findings may help to clarify the pathogenesis of adult-onset asthma.

Topics: Animals; Corticosterone; Cytokines; Female; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Lung; Lymph Nodes; Maternal Deprivation; Methacholine Chloride; Mice, Inbred BALB C; Mucus; Ovalbumin; Pneumonia; Respiratory Hypersensitivity; Stress, Psychological; T-Lymphocytes, Regulatory; Th2 Cells

The activation of Toll-like receptor (TLR) signaling is widely reported to be involved in preventing the development of allergic asthma. However, the mechanism of the protective function of TLR signaling remains limited. Here, we studied the mouse model of ovalbumin (OVA)-induced allergic asthma and found that deficiency of TLR signaling or activating TLR signaling with agonist would aggravate or attenuate OVA-induced allergic asthma, respectively, and TLR signaling-mediated protective effect mainly affected the sensitization phase. After OVA/alum sensitization, neutrophils and inflammatory monocytes were recruited into peritoneal cavity and up-regulated TLRs expression. However, adoptive transfer of inflammatory monocytes but not peritoneal macrophages or neutrophils induced allergic symptoms in recipient mice after OVA challenge even without OVA/alum sensitization, and treating the inflammatory monocytes with TLR agonist

Topics: Allergens; Animals; Asthma; Cell Movement; Cells, Cultured; Cytokines; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Inflammation Mediators; Mice; Mice, Inbred C57BL; Mice, Knockout; Monocytes; Neutrophils; Ovalbumin; Signal Transduction; Th1 Cells; Toll-Like Receptors

Kaempferol, a phytochemical found in many edible plants, is known to alleviate diseases such as cancer, allergy, and inflammation. The objective of this study was to investigate whether kaempferol could reduce omega-6 and ovalbumin-mediated allergic reactions at lung and trachea in BALB/c mice. Mice were allocated into five groups: 1) control group (CON); 2) positive control group with orally administration of omega-6 (POS); 3) bovine serum albumin (BSA) sensitization group (with BSA injection and ovalbumin inhalation); 4) BSA+K10 group: BSA injection, 10 μg/g of kaempferol administration and ovalbumin inhalation; and 5) BSA+K20 group: BSA injection, 20 μg/g of kaempferol administration and ovalbumin inhalation. Results revealed that serum histamine level was the highest (p<0.01) in BSA group. In lung tissue and trachea, cyclooxygenase 2 (Cox2) expression was significantly (p<0.05) higher in the BSA group compared to that in other groups. However, phosphorylated cytosolic phospholipase A2 (p-cPLA2) expression in the trachea was not significantly different among groups. Taken together, results of this study suggest that kaempferol might be useful for alleviating inflammation reaction associated with Cox2 expression. However, the exact mechanism of action involved in the effect of kaempferol on inflammatory response remains unclear.

Topics: Animals; Anti-Inflammatory Agents; Cyclooxygenase 2; Disease Models, Animal; Fatty Acids, Omega-6; Humans; Hypersensitivity; Kaempferols; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phospholipases A2, Cytosolic; Pneumonia; Trachea

Alstonia scholaris (L.) R. Br. (Apocynaceae), an important herbal medicine, has been widely used to treat respiratory tract diseases, such as cough, asthma, phlegm, and chronic obstructive pulmonary disease.. To evaluate pharmacological effect of alkaloids from A. scholaris on ovalbumin induced airways allergic inflammatory model, and explore whether the dosing frequency is related to pharmacokinetics.. After oral administration of total alkaloids, the pharmacokinetic study of it was investigated. In addition, anti-allergic studies were carried out on ovalbumin-sensitized airways allergic inflammatory model of mice.. The pharmacokinetics of total alkaloids (TA) was investigated in SD rat plasma by a fully-validated LC-MS/MS method. Then, an ovalbumin (OVA)-sensitized airways allergic inflammatory model was established, in which mice were intra-gastrically administrated by 3 times a day (8.3 and 16.7mg/kg) based on the pharmacokinetic behavior of TA) and single (25, 50mg/kg) treatment regimen. Dexamethasone was used as a positive control for corticosteroid drugs. Cellular infiltration was assessed in the broncho-alveolar lavage fluid (BALF). Expressions of interleukin-4 (IL-4) and interleukin-10 (IL-10) in the BALF were determined, levels of immunoglobulin E (IgE) and eotaxin in serum were measured, and superoxide dismutase (SOD) activities as well as malondialdehyde (MDA) content in the serum and BALF were examined. Finally, histopathological examination in the lung was assessed by H. E. staining.. The time course of plasma concentration of 4 bioactive indole alkaloids fitted an open two-compartment model after oral administration of total alkaloids at doses of 10, 25, and 50mg/kg. The area under the curve and the maximum concentration values of four major alkaloids increased dose-dependently, and half-life suggested a short-lasting pharmacological effect. Then, an ovalbumin-provoked airways allergic inflammatory model indicated that the pharmacological effect of administration of total alkaloids 3 times a day was a little better than that of single dose daily. The percentage of eosinophils in BALF was reduced obviously and the pathological damage of lung was also attenuated. There was also a significant reduction in IL-4 and promotion in IL-10 in the BALF. Serum IgE and eotaxin expression also significantly decreased in treated animals. Furthermore, the activity of SOD elevated remarkably and lipid peroxidation product (MDA) decreased in the administrated mice.. The pharmacological effects administrated for 3 times a day had precedence over single dose daily, which was related to the prolonged retention time and the maintained plasma concentration. Moreover, scholaricine and vallesamine might be responsible for the treatment of allergic asthma, mainly in total alkaloids.

Topics: Alkaloids; Alstonia; Animals; Anti-Allergic Agents; Drugs, Chinese Herbal; Eosinophils; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Rats; Rats, Sprague-Dawley

Anaphylactoid reactions, accounting for more than 77% of all immune-mediated immediate hypersensitivity reactions, have become a serious threat to public health, but their effect mechanism is not clear and diagnostic tests are limited. Comprehensive metabolite analysis may reveal the anaphylactoid effect mechanism systematically and provide reference for future diagnostic purposes.. Plasma from Brown Norway rats given intravenous injection of saline, compound 48/80 (2.5 mL/kg) or ovalbumin (20 mL/kg) in 20 s for the first time was used to study the effect mechanism of anaphylactoid reactions through metabolomics (UPLC-qTOF-MS/MS). Metabolomics integrated with proteomics data were used to analyze the anaphylactoid pathways by MetaboAnalyst followed by integrated pathway analysis.. Thirty metabolites were identified through the METLIN database by MS/MS and 18 of them were confirmed by authentic standards. The results showed that adenosine, histamine, N-acetylhistamine, N(α)-γ-glutamylhistamine, malate and xanthine are important indices for anaphylactoid reactions. It could be concluded that the effect mechanism is mainly composed of histidine metabolism, arachidonic acid metabolism, energy metabolism, purine metabolism and other small molecules through 30 metabolites. Multiple linear regression analysis indicated that not only histamine but also N(α)-γ-glutamylhistamine and arachidonic acid could be used to evaluate anaphylactoid symptoms of animals. Furthermore, the citrate cycle, histidine metabolism and arachidonic acid metabolism could be the main pathways of anaphylactoid reactions as determined by MetaboAnalyst.. The results may provide a reference to improve diagnostic accuracy and predict and monitor treatment efficacy in anaphylactoid reactions in the clinical setting.

Topics: Allergens; Anaphylaxis; Animals; Arachidonic Acid; Citric Acid; Disease Models, Animal; Histamine; Humans; Hypersensitivity; Male; Metabolomics; Ovalbumin; Proteomics; Rats; Rats, Inbred BN; Signal Transduction; Tandem Mass Spectrometry

Recently the role of gastrointestinal nematodes in modulating the immune responses in inflammatory and immune-mediated conditions such as allergy and autoimmune diseases has been introduced. This is mainly due to the suppressive effects of somatic and excretory secretory (ES) products of nematodes on the immune responses. In this study, we evaluated the immunomodulatory potentials of somatic products of Marshallagia marshalli, a gastrointestinal nematodes of sheep, to suppress the immune-mediated responses in a murine model of allergic airway inflammation. BALB/c mice were intraperitoneally (IP) sensitized with ovalbumin (OVA)/Alum and then challenged with 1% OVA. Somatic products of M. marshalli were administered during each sensitization. The effects of somatic products on development of allergic airway inflammation were evaluated by analyzing inflammatory cells recruitment, histopathological changes, cytokines production (IL-4, IL-13, IL-10, TGF-β) and serum antibody titers (IgG1, IgG2a).. Somatic products of M. marshalli were able to suppress the induction of allergic airway inflammation in mice. Modulation of Th2 type responses (IL-4, IL-13, IgG1) via upregulations of IL-10 and TGF-β production was observed after injection of somatic products of M. marshalli. In addition, inflammatory cells infiltration and pathological disorders were significantly diminished following administration of somatic products.. Our data raised the possibility that helminths could be a potential therapeutic candidate to alleviate the inflammatory conditions in allergic asthma. According to these results, we concluded that M. marshalli may contain immune-modulatory molecules that attenuate allergic airway inflammation via induction of regulatory cytokines. Further investigations are required to identify molecules that might have potentials for development of novel therapeutic targets.

Topics: Alum Compounds; Animals; Asthma; Cytokines; Disease Models, Animal; Down-Regulation; Hypersensitivity; Immunization; Immunomodulation; Interleukin-10; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Sheep; Th2 Cells; Tissue Extracts; Trichostrongyloidea

Pulmonary neuroendocrine cells (PNECs) are the only innervated airway epithelial cells. To what extent neural innervation regulates PNEC secretion and function is unknown. Here, we discover that neurotrophin 4 (NT4) plays an essential role in mucus overproduction after early life allergen exposure by orchestrating PNEC innervation and secretion of GABA. We found that PNECs were the only cellular source of GABA in airways. In addition, PNECs expressed NT4 as a target-derived mechanism underlying PNEC innervation during development. Early life allergen exposure elevated the level of NT4 and caused PNEC hyperinnervation and nodose neuron hyperactivity. Associated with aberrant PNEC innervation, the authors discovered that GABA hypersecretion was required for the induction of mucin Muc5ac expression. In contrast,

Topics: Allergens; Animals; Calcium; gamma-Aminobutyric Acid; Gene Expression Regulation; Hypersensitivity; Mice, Inbred C57BL; Mucus; Neuroendocrine Cells; Ovalbumin

The neutrophil-activating protein of Helicobacter pylori (HP-NAP) has been identified as a modulator with anti-Th2 inflammation activity, and cholera toxin B (CTB) is a mucosal adjuvant that can also induce antigen tolerance. In this study, we constructed a CTB-NAP fusion protein on the surface of Bacillus subtilis spore and evaluate the efficiency of oral administration of the recombinant CTB-NAP spores in preventing asthma in mice. Oral administration of recombinant CTB or CTB-NAP spores significantly decreased serum ovalbumin (OVA)-specific IgE (p < 0.001) and increased fecal IgA (p < 0.01) compared to the treatment with non-recombinant spores. Oral administration of recombinant CTB or CTB-NAP spores induced IL-10 and IFN-γ expression and reduced IL-4 levels in bronchoalveolar lavage fluid (BALF). Moreover, CTB and CTB-NAP spores reduced the eosinophils in BALF and inflammatory cell infiltration in the lungs. Furthermore, CD4

Topics: Adjuvants, Immunologic; Administration, Oral; Animals; Asthma; Bacillus subtilis; Bacterial Proteins; Cholera Toxin; Hypersensitivity; Immunoglobulin E; Inflammation; Interferon-gamma; Interleukin-10; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Fusion Proteins; Spores, Bacterial; T-Lymphocytes, Regulatory

Topics: Animals; Antibodies; Bronchial Hyperreactivity; CD4-Positive T-Lymphocytes; Cell Differentiation; Cell Polarity; Female; Hypersensitivity; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Pneumonia; Receptors, Tumor Necrosis Factor, Type II; Signal Transduction; Th17 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

Asthma is characterized by intermittent airway obstruction and chronic inflammation, orchestrated primarily by Th2 cytokines. There is a strong rationale for developing new asthma therapies, since current treatment protocols present side effects and may not be effective in cases of difficult-to-control asthma. The purpose of this study was to evaluate the effect of ferulic acid, a phenolic acid commonly present in plants, in the ovalbumin-induced pulmonary allergy murine model.. BALB/c mice were sensitized and challenged with ovalbumin, and treatments were provided by gavage. Six groups of mice (n = 6) were studied, labeled as: control, pulmonary allergy, dexamethasone, and 3 receiving ferulic acid (at 25, 50, and 100 mg/kg). Lung tissue, bronchoalveolar lavage fluid and serum were collected for analysis.. Ferulic acid treatment inhibited an established allergic Th2-response by decreasing the key features of pulmonary allergy, including lung and airway inflammation, eosinophil infiltration, mucus production and serum levels of OVA-specific IgE. These results were associated with lower levels of CCL20, CCL11 and CCL5 chemokines and IL-4, IL-5, IL-13, TSLP, IL-25 and IL-33 cytokines in lung tissue homogenate.. In this study it was demonstrated for the first time that ferulic acid treatment is able to suppress one of the main features of the airway remodeling, indicated by reduction of mucus production, besides the Th2 pathogenic response on ovalbumin-induced pulmonary allergy. Taken together, results shows that the immunopathological mechanism underlying these effects is linked to a reduction of the epithelial-derived chemokines and cytokines, suggesting that ferulic acid may be useful as a potential therapeutic agent for asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Chemokines; Coumaric Acids; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Triticum aestivum sprouts are small shoots that germinate from seeds and are consumed as a dietary supplement. The present study aimed to determine whether a dichloromethane fraction isolated from Triticum aestivum sprouts (TDF) suppressed the allergic immune response in ovalbumin (OVA)‑sensitized mice. In vivo experiments were performed by administering TDF or vehicle to mice during the sensitization and this was immediately followed by an intradermal injection of OVA into the ears. Splenocytes isolated from OVA‑sensitized mice were pre‑treated with TDF and re‑challenged with OVA for ex vivo evaluation. Results demonstrated that TDF suppressed the inflammatory response in ear tissues and levels of total immunoglobulin (Ig)E and OVA‑specific IgE in serum. TDF inhibited the production of interleukin (IL)‑4 and expression of GATA‑binding protein‑3 (GATA‑3) transcription factor which regulates the differentiation of naïve T helper (Th) cells into Th2 cells in OVA‑stimulated splenocytes. TDF inhibited Th1‑associated cytokine interferon‑γ and IL‑12 production and downregulated the expression of Th1 specific transcription factor T‑box 21 in OVA‑stimulated splenocytes. Overall, these results indicated that TDF attenuates OVA‑induced allergic immune response by suppressing the production of Th2 specific cytokine IL‑4, through inhibiting transcription factor GATA‑3, and suggests that TDF may exhibit the potential to regulate the immune response in allergic diseases.

Topics: Animals; Cell Differentiation; Cell Separation; Cytokines; Female; Hypersensitivity; Immunity; Immunization; Immunoglobulins; Methylene Chloride; Mice, Inbred BALB C; Ovalbumin; Skin; Spleen; Th1 Cells; Th2 Cells; Transcription Factors; Triticum

Diospyros kaki L. (Ebenaceae) fruit is widely distributed in Asia and is known to exert anti-inflammatory and antithrombotic effects.. We evaluated the inhibitory effect of aqueous extract of D. kaki calyx (AEDKC) on mast cell-mediated immediate-type hypersensitivity and underlying mechanism of action.. For in vivo, ovalbumin (OVA)-induced active systemic anaphylaxis (ASA) and immunoglobulin (Ig) E-mediated passive cutaneous anaphylaxis (PCA) models were used. In the ASA, AEDKC (1-100 mg/kg) was orally administered 3 times during 14 days. In the PCA, AEDKC was orally treated 1 h before the antigen challenge. The control drug dexamethasone was used to compare the effectiveness of AEDKC. For in vitro, IgE-stimulated RBL-2H3 cells and primary cultured peritoneal mast cells were used to determine the role of AEDKC (0.01-1 mg/mL).. Oral administration of AEDKC dose dependently suppressed rectal temperature decrease and increases in serum histamine, total IgE, OVA-specific IgE, and interleukin (IL)-4 in the ASA. In the PCA, AEDKC reduced Evans blue pigmentation. Compared to dexamethasone (10 mg/kg), AEDKC (100 mg/kg) showed similar inhibitory effects in vivo. AEDKC concentration dependently suppressed the release of histamine and β-hexosaminidase through the reduction of intracellular calcium in mast cells. In addition, AEDKC decreased the expression and secretion of tumour necrosis factor-α and IL-4 by the reduction of nuclear factor-κB. The inhibitory potential of AEDKC (1 mg/mL) was similar with dexamethasone (10 μM) in vitro.. We suggest that AEDKC may be a potential candidate for the treatment of mast cell-mediated allergic diseases.

Topics: Anaphylaxis; Animals; Cell Survival; Cells, Cultured; Diospyros; Dose-Response Relationship, Drug; Hypersensitivity; Male; Mast Cells; Mice; Mice, Inbred ICR; Ovalbumin; Plant Extracts; Rats; Rats, Sprague-Dawley

Pidotimod is a synthetic dipeptide with biological and immuno‑modulatory properties. It has been widely used for treatment and prevention of recurrent respiratory infections. However, its impact on the regulation of allergic pulmonary inflammation is still not clear. In the current study, an ovalbumin (OVA)‑induced allergic asthma model was used to investigate the immune‑modulating effects of pidotimod on airway eosinophilia, mucus metaplasia and inflammatory factor expression compared with dexamethasone (positive control). The authors determined that treatment with pidotimod exacerbated pulmonary inflammation as demonstrated by significantly increased eosinophil infiltration, dramatically elevated immunoglobulin E production, and enhanced T helper 2 response. Moreover, treatment failed to attenuate mucus production in lung tissue, and did not reduce OVA‑induced high levels of FIZZ1 and Arg1 expression in asthmatic mice. In contrast, administration of dexamethasone was efficient in alleviating allergic airway inflammation in OVA‑induced asthmatic mice. These data indicated that pidotimod as an immunotherapeutic agent should be used cautiously and the effectiveness for controlling allergic asthma needs further evaluation and research.

Topics: Animals; Arginase; Asthma; Bronchoalveolar Lavage Fluid; Cell Differentiation; Disease Models, Animal; Down-Regulation; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Intercellular Signaling Peptides and Proteins; Lung; Metaplasia; Mice, Inbred C57BL; Mucus; Ovalbumin; Pyrrolidonecarboxylic Acid; Respiratory Tract Infections; Th2 Cells; Thiazolidines

Natural products have a prime importance as an essential source for new drug discovery. Carica papaya leaves (CPL) have been used to treat inflammation in traditional system of medicine.. Current study evaluates the anti-inflammatory and immunomodulatory effects of CPL extract using mouse model of ovalbumin- (OVA) induced allergic asthma.. All the mice were intraperitoneally sensitized and subsequently given intranasal challenge with OVA except the control group. Group-III and -IV were treated for seven consecutive days with CPL extract and methylprednisolone (MP), respectively. At the end of study, histopathological examination of the lungs was performed and inflammatory cell counts were done in blood as well as bronchoalveolar lavage fluid (BALF). The mRNA expression levels of IL-4, IL-5, eotaxin, TNF-α, NF-ĸB, and iNOS were measured using reverse transcription polymerase chain reaction (RT-PCR).. Results showed significant attenuation of lung infiltration of inflammatory cells, alveolar thickening, and goblet cell hyperplasia after treatment with CPL extract. We also found significant suppression of total and differential leukocyte counts in both blood and BALF samples of CPL extract treated group. CPL extract also alleviated the expression levels of IL-4, IL-5, eotaxin, TNF-α, NF-ĸB, and iNOS. Similarly, treatment with MP, used as a reference drug, also significantly ameliorated all the pro-inflammatory markers.. Current study shows that CPL extract possesses anti-inflammatory effect in mouse model of allergic airway inflammation by down-regulating IL-4, IL-5, eotaxin, TNF-α, NF-ĸB, and iNOS expression levels.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Bronchoalveolar Lavage Fluid; Carica; Disease Models, Animal; Down-Regulation; Hypersensitivity; Interleukin-4; Interleukin-5; Lung; Male; Mice, Inbred BALB C; NF-kappa B; Nitric Oxide Synthase Type II; Ovalbumin; Tumor Necrosis Factor-alpha

Asthma is associated with exposure to a wide variety of allergens and adjuvants. The extent to which overlap exists between the cellular and molecular mechanisms triggered by these various agents is poorly understood, but it might explain the differential responsiveness of patients to specific therapies. In particular, it is unclear why some, but not all, patients benefit from blockade of TNF. Here, we characterized signaling pathways triggered by distinct types of adjuvants during allergic sensitization. Mice sensitized to an innocuous protein using TLR ligands or house dust extracts as adjuvants developed mixed eosinophilic and neutrophilic airway inflammation and airway hyperresponsiveness (AHR) following allergen challenge, whereas mice sensitized using proteases as adjuvants developed predominantly eosinophilic inflammation and AHR. TLR ligands, but not proteases, induced TNF during allergic sensitization. TNF signaled through airway epithelial cells to reprogram them and promote Th2, but not Th17, development in lymph nodes. TNF was also required during the allergen challenge phase for neutrophilic and eosinophilic inflammation. In contrast, TNF was dispensable for allergic airway disease in a protease-mediated model of asthma. These findings might help to explain why TNF blockade improves lung function in only some patients with asthma.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Cell Differentiation; Cytokines; Disease Models, Animal; Eosinophils; Hypersensitivity; Inflammation; Interleukin-17; Ligands; Lung; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Ovalbumin; Respiratory Hypersensitivity; Signal Transduction; Th17 Cells; Th2 Cells; Toll-Like Receptors; Tumor Necrosis Factor-alpha

Allergy cases are increasing worldwide. Currently allergies are treated after their appearance in patients. However, now there is effort to make a preventive vaccine against allergies. The rationale is to target patient populations that are already sensitized to allergens but have yet to develop severe forms of the allergic disease, or who are susceptible to allergy development but have not yet developed them. Subcutaneous injections and the sublingual route have been used as the primary mode of preventive vaccine delivery. However, injections are painful, especially considering that they have to be given repeatedly to infants or young children. The sublingual route is hard to use since infants can't be trained to hold the vaccine under their tongue. In the present study, we demonstrate a microneedle (MN)-based cutaneous preventive allergy treatment against ovalbumin (Ova)-induced airway allergy in mice. Insertion of MNs coated with Ova as a model allergen and CpG oligonucleotide as an adjuvant (MNs-CIT) into the skin significantly induced Ova specific systemic immune response. This response was similar to that induced by hypodermic-needle-based delivery of Ova using the clinically-approved subcutaneous immunotherapy (SCIT) route. MNs-CIT regulated Th2 cytokines (IL-4, IL-5 & IL-13) and anti-inflammatory cytokines (IL-10) in the bronchoalveolar fluid, and IL-2 and IFN-γ cytokines in restimulated splenocyte cultures. Absence of mucus deposition inside the bronchiole wall and low collagen around the lung bronchioles after Ova-allergen challenge further confirmed the protective role of MNs-CIT. Overall, MNs-CIT represents a novel minimally invasive cutaneous immunotherapy to prevent the progression of Ova induced airway allergy in mice.

Topics: Adjuvants, Immunologic; Administration, Cutaneous; Airway Obstruction; Allergens; Animals; Cell Line; Drug Delivery Systems; Female; Hypersensitivity; Immunotherapy; Interleukins; Mice, Inbred BALB C; Microinjections; Needles; Oligodeoxyribonucleotides; Ovalbumin; Skin; Vaccination; Vaccines

Oxidative stress is important in the pathogenesis of allergic asthma. Extracellular superoxide dismutase (EC-SOD; SOD3) is the major antioxidant in lungs, but its role in allergic asthma is unknown. Here we report that asthmatics have increased SOD3 transcript levels in sputum and that a single nucleotide polymorphism (SNP) in SOD3 (R213G; rs1799895) changes lung distribution of EC-SOD, and decreases likelihood of asthma-related symptoms. Knockin mice analogous to the human R213G SNP had lower airway hyperresponsiveness, inflammation, and mucus hypersecretion with decreased interleukin-33 (IL-33) in bronchoalveolar lavage fluid and reduced type II innate lymphoid cells (ILC2s) in lungs. SOD mimetic (Mn (III) tetrakis (N-ethylpyridinium-2-yl) porphyrin) attenuated Alternaria-induced expression of IL-33 and IL-8 release in BEAS-2B cells. These results suggest that R213G SNP potentially benefits its carriers by resulting in high EC-SOD in airway-lining fluid, which ameliorates allergic airway inflammation by dampening the innate immune response, including IL-33/ST2-mediated changes in ILC2s.

Topics: Aged; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cohort Studies; Cytokines; Female; Genotype; Humans; Hypersensitivity; Immunity, Innate; Interleukin-33; Lymphocytes; Male; Mice; Mice, Transgenic; Middle Aged; Ovalbumin; Phenotype; Polymorphism, Single Nucleotide; Porphyrins; Respiratory Hypersensitivity; RNA, Messenger; Sputum; Superoxide Dismutase

Allergy is an acquired hypersensitivity reaction of the immune system mediated by IgE‑induced mast cell degranulation. In China, bacillus Calmette‑Guerin extract (BCGE) has been shown to be clinically effective for regulating immunity, which enhances the resistance of the body to anaphylactic disease, infectious diseases and cancer. However, the mechanisms remain to be fully elucidated. The present study investigated the potential anti‑allergic effects of BCGE in animal models of mast cell‑dependent anaphylaxis and mechanisms of BCGE in mast cells. Anti‑allergic actions of BCGE were evaluated in passive cutaneous anaphylaxis, dextran T40‑induced scratching behavior mouse models, and in ovalbumin (OVA)‑induced contraction of intestinal tube isolated from OVA‑sensitized guinea pigs. Direct mast cell‑stabilizing effects of BCGE were examined in mast cells from the abdominal cavity of OVA‑sensitized rats. Anti‑allergic signaling mechanisms of BCGE in mast cells were investigated by detection of cyclic adenosine monophosphate levels and protein kinase A expression in mast cells. It was observed that BCGE prevented OVA‑induced cutaneous vascular hyperpermeability, skin itching, elevation in plasma histamine levels and abdominal cavity fluid mast cell degranulation in animal models, in a dose‑dependent manner. BCGE also suppressed OVA‑mediated guinea pig intestinal tube contraction in vitro. In addition, BCGE was found to increase the levels of interferon‑γ, and reduce the levels of interleukin‑4 and OVA‑sIg E levels in OVA‑sensitized rats. BCGE also increased levels of cyclic adenosine monophosphate and the expression of protein kinase A in mast cells separated from the abdominal cavity fluid of OVA‑sensitized rats. In conclusion, the results suggested that BCGE possesses anti‑allergic activity by inhibiting IgE‑induced mast cell degranulation, providing a foundation for the development of BCGE for the treatment of mast cell‑mediated allergic disorders.

Topics: Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Bacillus; Cell Degranulation; Disease Models, Animal; Female; Guinea Pigs; Histamine; Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulin E; Mast Cells; Mice; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Wistar

Genipin is a natural compound isolated from the fruit of Gardenia jasminoides with various pharmacological effects. In this study, we investigated whether genipin effectively alleviates allergic responses in a murine model of ovalbumin (OVA)-induced asthma. The mice were administered an intraperitoneal injection of OVA on day 0 and 14 to boost the immune response; genipin was then administered from day 18 to 23 by oral gavage. On days 21 to 23, mice were OVA-challenged using am ultrasonic nebulizer, and airway hyperresponsiveness (AHR) was determined on day 24 by plethysmography. Genipin significantly reduced the inflammatory cell count in bronchoalveolar lavage fluids (BALF) and AHR, which were accompanied by lower interleukin-5 (IL-5), IL-13 and OVA-specific immunoglobulin (Ig) E levels in the BALF or serum from OVA-induced asthmatic mice. In histology, genipin significantly decreased airway inflammation and mucus hypersecretion in OVA-induced asthmatic mice. Additionally, genipin inhibited OVA-induced increases in the expression of inducible nitric oxide synthase and cyclooxygenase-2 proteins. Further, genipin reduced the activity and protein levels of matrix metalloproteinase-9 in lung tissue from OVA induced asthmatic mice. Overall, genipin effectively alleviated the asthmatic inflammatory response in an OVA-induced asthmatic model. Therefore, our results suggest that genipin has therapeutic potential for treating asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Gardenia; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-5; Iridoids; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Nitric Oxide Synthase Type II; Ovalbumin

Asthma is a chronic inflammation of the airways affecting over 300 million people worldwide. As in the autoimmune diseases, it is well described that women are the most affected by asthma. The higher number of women presenting this pathology suggests the involvement of female sex hormones in the construction of the allergic immune response. Female Balb / c mice were used for the experiments. Thirty-eight animals were separated into four groups: OVX-Ova; Sham-Ova; OVX-Sal; Sham-Sal. Then animals underwent acute allergic induction protocol by Ovalbulmin (OVA). Ovariectomized animals showed greater number of leukocytes in bronchoalveolar lavage (BAL) and elevated white blood cells recruitment to the lung environment observed by histological analysis. There was a significant increase of eosinophils and mast cells in inflammatory sites at pulmonary tissue. The relative uterine and body weight were lower in ovariectomized animals and higher in Sham mice, respectively. Moreover, the lack of the sex hormones induced an increase in interleukin (IL)-4 and titers of immunoglobulin G1 (IgG1) antibodies. However, increased production of IL-17A was only observed in Sham animals. Altogether, data this study suggest that ovariectomy induces the formation of a stronger Th2 response in allergic animal. However, the immune processes involved in the allergic response in females currently remain unclear.

Topics: Animals; Antibodies; Chemotaxis, Leukocyte; Cytokines; Eosinophils; Female; Gonadal Steroid Hormones; Hypersensitivity; Immunoglobulin G; Lung; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Ovariectomy; Th17 Cells; Th2 Cells

Respiratory viral infections have frequently been reported to closely correlate with asthma exacerbations. Distinctive expression of cytokine/chemokine and anomalous responses of innate immunity induced by respiratory viral infections were suggested to play a key role. This study further evaluates the effects of airway sensitization on innate immunity in response to different viruses.. Murine sensitization was established using an ovalbumin (OVA) sensitization model. Mice were subsequently infected with either respiratory syncytial virus (RSV) or human metapneumovirus (hMPV). Type I interferon (IFN), cytokines, and chemokines were measured in bronchoalveolar lavage (BAL) fluid. Pulmonary tissue samples were collected for the analysis of viral titers and type I IFN signal transcriptors.. Distinct expressions of cytokine/chemokine responses after viral infection were also found in mice with OVA sensitization. A significant increase of virus replication was found in lungs of RSV-infected sensitized mice. The increment of RSV titer was associated with the decreased levels of type I IFN. Although Toll-like receptor 3 (TLR3) expression was significantly increased in the lungs, the key signal transcriptor, IFN regulatory factor 3, was significantly suppressed in the RSV-infected sensitized mice.. A defective antiviral innate response was observed in the murine respiratory allergy model. Suppressed expression of IFN signal transcriptor contributes to decreased production of type I IFN. The defective innate immune response might result in acute viral exacerbations of allergic airway diseases.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Disease Susceptibility; Female; Hypersensitivity; Immunity, Innate; Lung; Mice, Inbred BALB C; Ovalbumin; Paramyxoviridae Infections; Respiratory Syncytial Virus Infections

Allergic asthma is a chronic inflammatory disorder of the airways that affects >300 million people worldwide. The pro-inflammatory cytokines interleukin (IL)-1α and IL-1β have essential roles in the pathogenesis of asthma. However, the mechanisms underlying the production of IL-1 cytokines in allergic asthma remain unclear. In this study, we used a mouse model of ovalbumin-induced asthma to identify a crucial role for caspase-8 in the development of allergic airway inflammation. We further demonstrated that hematopoietic cells have dominant roles in caspase-8-mediated allergic airway inflammation. Caspase-8 was required for the production of IL-1 cytokines to promote Th2 immune response, which promotes the development of pulmonary eosinophilia and inflammation. Thus, our study identifies caspase-8 as a master regulator of IL-1 cytokines that contribute to the pathogenesis of asthma and implicates caspase-8 inhibition as a potential therapeutic strategy for asthmatic patients.

Topics: Allergens; Animals; Asthma; Caspase 8; Cells, Cultured; Disease Models, Animal; Humans; Hypersensitivity; Interleukin-1; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Signal Transduction; Th2 Cells

Sublingual immunotherapy (SLIT) is a safe and efficient treatment for type 1 allergies; however, the underlying immunological mechanisms, particularly the phenotype of oral antigen-presenting cells (APCs) responsible for the induction of regulatory T (Treg) cells, remain unclear. We show here that the sublingual application of ovalbumin (OVA) induced antigen-specific Foxp3

Topics: Animals; Antigen Presentation; Antigens; Antigens, CD; CD11b Antigen; Cell Differentiation; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Forkhead Transcription Factors; Humans; Hypersensitivity; Integrin alpha Chains; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Sublingual Immunotherapy; T-Lymphocytes, Regulatory

Immunomodulatory interventions play a key role in the treatment of infections and cancer as well as allergic diseases. Adjuvants such as micro- and nanoparticles are often added to immunomodulatory therapies to enhance the triggered immune response. Here, we report the immunological assessment of novel and economically manufactured microparticle adjuvants, namely strontium-doped hydroxyapatite porous spheres (SHAS), which we suggest for the use as adjuvant and carrier in allergen-specific immunotherapy (ASIT).. Scanning electron microscopy revealed that the synthesis procedure developed for the production of SHAS results in a highly homogeneous population of spheres. Strontium-doped hydroxyapatite porous spheres bound and released proteins such as ovalbumin (OVA) or the major cat allergen Fel d 1. SHAS-OVA were taken up by human monocyte-derived dendritic cells (mdDCs) and murine DCs and did not have any necrotic or apoptotic effects even at high densities. In a murine model of ASIT for allergic asthmatic inflammation, we found that OVA released from subcutaneously injected SHAS-OVA led to a sustained stimulation of both CD4. We conclude that SHAS may constitute a suitable carrier and adjuvant for ASIT with great potential due to its unique protein-binding properties.

Topics: Adjuvants, Immunologic; Allergens; Animals; Dendritic Cells; Desensitization, Immunologic; Disease Models, Animal; Female; Hydroxyapatites; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Lymphocyte Activation; Mice; Ovalbumin; Phosphatidylethanolamines; Strontium; T-Lymphocyte Subsets; Treatment Outcome

To understand mechanisms of human olfactory dysfunction in chronic rhinosinusitis, an inducible olfactory inflammation (IOI) model has been utilized to chronically express inflammatory cytokines locally, resulting in neuronal loss, diminished odorant responses, and repressed olfactory regeneration. Knockout of the minor tumor necrosis factor α receptor 2 (TNFR2) was previously shown to partially rescue these olfactory changes. The purpose of current study was to investigate the role of the major TNF receptor, TNFR1, in chronic olfactory inflammation.. Two experimental groups of mice were studied: TNFR1 knockout in IOI background and TNFR1 knockout with allergen-induced inflammation. Olfactory function was assayed by electro-olfactogram (EOG), and olfactory tissue was processed for histology and immunohistochemical staining.. TNF-α was dramatically induced in IOI-TNFR1 knockout mice, but the olfactory epithelium did not show inflammation. EOG responses were normal after either 2 or 8 weeks of TNF-α expression. Ovalbumin-sensitized TNFR1 knockout mice developed markedly diminished eosinophilic inflammatory infiltration.. Genetic deletion of TNFR1 completely blocks TNF-α-induced inflammation and reduces allergen-induced inflammation. Preserved EOG responses suggest a TNFR1-dependent mechanism of TNF-α-induced olfactory neuron dysfunction.

Topics: Allergens; Alum Compounds; Animals; Female; Hypersensitivity; Inflammation; Male; Mice, Knockout; Nasal Lavage Fluid; Neurons; Olfaction Disorders; Olfactory Mucosa; Ovalbumin; Receptors, Tumor Necrosis Factor, Type I; Tumor Necrosis Factor-alpha

Pruritus is the major symptom of ocular allergy but currently available treatments are often ineffective. Previous studies demonstrated that subpopulations of primary sensory neurons express Fc receptors and may contribute to antigen-specific pain. We investigated the role of neuronal Fc-epsilon Receptor I (FcεRI) in allergic ocular pruritus. Ovalbumin (OVA) was used as allergen together with alum adjuvant (OVA+alum) to produce a mouse model of ocular allergy with a significant elevation in the serum levels of both antigen-specific IgE and IgG. Mice sensitized by OVA without alum only induced elevation of serum IgG but not IgE. Scratching behavior toward the eyes with the hindlimb was used as an indicator of ocular itch. Topical OVA challenging to the eye dose-dependently induced scratching toward the eye in the OVA+alum sensitized mice, but not those sensitized by OVA only. The antigen-induced scratching was largely abolished by topical application of the blocking antibody to FcεRIα, but was only partially alleviated by pretreatment of mast cell stabilizer or histamine I receptor antagonist. The expression of FcεRI was detected in subpopulations of trigeminal ganglion (TG) neurons including those expressing pruriceptive markers and innervating the conjunctiva in the naïve mice. Moreover, FcεRI was found significantly upregulated in small-sized TG neurons in the OVA+alum sensitized mice. In acutely dissociated TG neurons, IgE-immune complex (IC), but not the antibody or antigen alone, induced intracellular calcium increase. The neuronal responses to IgE-IC could be specifically blocked by pre-application of a siRNA for FcεRIα. Our results indicate that FcεRI expressed on peripheral nociceptive neurons in the TG may be directly activated by IgE-IC and contribute to allergic ocular pruritus. This study may suggest a novel mechanism for the development of pathological itch in allergic diseases.

Topics: Alum Compounds; Animals; Disease Models, Animal; Eye Diseases; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Neurons; Ovalbumin; Pruritus; Receptors, IgE

Rho kinases (ROCKs) contribute to allergic airways disease. ROCKs also play a role in lymphocyte proliferation and migration.. To determine the role of ROCK2 acting within CD4. ROCK2-haploinsufficient (ROCK2. OVA-induced increases in bronchoalveolar lavage lymphocytes, eosinophils, IL-13, IL-5, and eotaxin were reduced in ROCK2. ROCK2 contributes to allergic airways responses likely via effects within ASM cells and within non-lymphocyte cells involved in lymphocyte activation and migration into the airways.

Topics: Adoptive Transfer; Animals; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Disease Models, Animal; Gene Deletion; Goblet Cells; Hypersensitivity; Male; Mice; Mice, Knockout; Ovalbumin; Respiratory Mucosa; rho-Associated Kinases; T-Cell Antigen Receptor Specificity

Mast cells are important effector cells in immunoglobulin (Ig) E-mediated allergic reactions such as asthma, atopic dermatitis and rhinitis. Vanillic acid, a natural product, has shown anti-oxidant and anti-inflammatory activities. In the present study, we investigated the anti-allergic inflammatory effects of ortho-vanillic acid (2-hydroxy-3-methoxybenzoic acid, o-VA) that was a derivative of vanillic acid isolated from Amomum xanthioides. In mouse anaphylaxis models, oral administration of o-VA (2, 10, 50 mg/kg) dose-dependently attenuated ovalbumin-induced active systemic anaphylaxis and IgE-mediated cutaneous allergic reactions such as hypothermia, histamine release, IgE production and vasodilation; administration of o-VA also suppressed the mast cell degranulator compound 48/80-induced anaphylaxis. In cultured mast cell line RBL-2H3 and isolated rat peritoneal mast cells in vitro, pretreatment with o-VA (1-100 μmol/L) dose-dependently inhibited DNP-HSA-induced degranulation of mast cells by decreasing the intracellular free calcium level, and suppressed the expression of pro-inflammatory cytokines TNF-α and IL-4. Pretreatment of RBL-2H3 cells with o-VA suppressed DNP-HSA-induced phosphorylation of Lyn, Syk, Akt, and the nuclear translocation of nuclear factor-κB. In conclusion, o-VA suppresses the mast cell-mediated allergic inflammatory response by blocking the signaling pathways downstream of high affinity IgE receptor (FcεRI) on the surface of mast cells.

Topics: Anaphylaxis; Animals; Benzoates; Calcium; Cell Degranulation; Cells, Cultured; Dinitrophenols; Dose-Response Relationship, Drug; Hypersensitivity; Immunoglobulin E; Inflammation Mediators; Male; Mast Cells; Mice; NF-kappa B; Ovalbumin; p-Methoxy-N-methylphenethylamine; Phosphorylation; Rats; Receptors, IgE; Serum Albumin; Signal Transduction; Vanillic Acid

Trigonella foenum-graecum, a member oldest medicinal plant in the fabaceae (legumes) family, is used as a herb, spice, and vegetable, and known for its olfactory, laxative, and galactogogue effects. However, the inhibitory effect of Trigonella foenum-graecum on allergic inflammatory response remains unclear, therefore, we investigated the precise role of Trigonella foenum-graecum in the allergic asthma and revealed the effects of Trigonella foenum-graecum in regulating airway inflammation and its possible mechanism. Allergic asthma was initiated in BALB/c mice by sensitized with OVA emulsified in aluminum on days 1 and 14, then aerosol challenged with OVA on days 27, 28 and 29. Some mice were administered Trigonella foenum-graecum by oral gavage before challenge. Then mice were evaluated for the presence of airway inflammation, production of allergen-specific cytokine response and lung pathology. Trigonella foenum-graecum significantly ameliorated the number of inflammatory cells in BALF and alleviated lung inflammation. It also reduced the collagen deposition and goblet cells. Meanwhile, Trigonella foenum-graecum treatment evidently decreased the high expression of Th2 cytokines and increased the Th1 cytokines in BALF and lung homogenates. Trigonella foenum-graecum showed a significant inhibition of serum IgE and anti-OVA IgG

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Th2 Cells; Trigonella

Allergic asthma, caused by inhaled allergens such as house dust mite or grass pollen, is characterized by reversible airway obstruction, associated with an eosinophilic inflammation of the airways, as well as airway hyper responsiveness and remodeling. The inhaled allergens trigger a type-2 inflammatory response with involvement of innate lymphoid cells (ILC2) and Th2 cells, resulting in high production of immunoglobulin E (IgE) antibodies. Consequently, renewed allergen exposure results in a classic allergic response with a distinct early and late phase, both resulting in bronchoconstriction and shortness of breath. Allergen specific immunotherapy (AIT) is the only treatment that is capable of modifying the immunological process underlying allergic responses including allergic asthma and both subcutaneous AIT (SCIT) as well as sublingual AIT (SLIT) have proven clinical efficacy in long term suppression of the allergic response. Although these treatments are very successful for rhinitis, application of AIT in asthma is hampered by variable efficacy, long duration of treatment, and the risk of severe side-effects. A more profound understanding of the mechanisms by which AIT achieves tolerance to allergens in sensitized individuals is needed to improve its efficacy. Mouse models have been very valuable as a preclinical model to characterize the mechanisms of desensitization in AIT and to evaluate novel approaches for improved efficacy. Here, we present a rapid and reproducible mouse model for allergen-specific immunotherapy. In this model, mice are sensitized with two injections of allergen absorbed to aluminum hydroxide to induce allergic sensitization, followed by subcutaneous injections (SCIT) or sublingual administrations (SLIT) of the allergen as immunotherapy treatment. Finally, mice are challenged by three intranasal allergen administrations. We will describe the protocols as well as the most important read-out parameters including measurement of invasive lung function measurements, serum immunoglobulin levels, isolation of broncho-alveolar lavage fluid (BALF), and preparation of cytospins. Moreover, we describe how to restimulate lung single cell suspensions, perform flow cytometry measurements to identify populations of relevant immune cells, and perform ELISAs and Luminex assays to measure the cytokine concentrations in BALF and lung tissue.

Topics: Allergens; Animals; Antigens, Plant; Asthma; Bronchoalveolar Lavage Fluid; Complex Mixtures; Cytokines; Disease Models, Animal; Female; Flow Cytometry; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Injections, Subcutaneous; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Pyroglyphidae; Sublingual Immunotherapy

Schisandra chinensis (SC) and its main constituent, schizandrin (SCH) exhibit anti-inflammatory and anti-allergic activities. Allergic and inflammatory reactions are aggravated via caspase-1 signaling pathway. However, the regulatory effects of SC and SCH on caspase-1 activation have not been clarified yet. In this study, we aimed to clarify the anti-allergic effects of SC and SCH using an ovalbumin (OVA)-sensitized mice and anti-CD3 and anti-CD28 antibodies-stimulated splenocytes. SC or SCH significantly inhibited the levels of immunoglobulin (Ig)E, IgG1, or interleukin (IL)-4 in serum of OVA-sensitized mice. SC or SCH significantly inhibited the levels of IL-6, tumor necrosis factor (TNF)-[Formula: see text], and IL-1[Formula: see text] in spleen of the OVA-sensitized mice. SC or SCH significantly suppressed the expression of caspase-1 and receptor-interacting protein (RIP)-2 in spleen of the OVA-sensitized mice. In activated splenocytes, SC or SCH significantly decreased the expression of caspase-1 and RIP-2 as well as the production of IL-6 and TNF-[Formula: see text]. We suggest that SC and SCH exert an anti-allergic effect by down-regulating caspase-1 signaling.

Topics: Animals; Caspase 1; Cyclooctanes; Down-Regulation; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-1beta; Interleukin-4; Interleukin-6; Lignans; Mice; Ovalbumin; Plant Extracts; Polycyclic Compounds; Receptor-Interacting Protein Serine-Threonine Kinase 2; Receptor-Interacting Protein Serine-Threonine Kinases; Schisandra; Spleen; Tumor Necrosis Factor-alpha

Previous epidemiologic studies have demonstrated an inverse association between Helicobacter pylori infection and the frequency of allergic asthma. The neutrophil-activating protein (NAP) of H. pylori has been identified as a modulator possessing anti-Th2 inflammation activity. Here, we sought to determine whether systemic or mucosal pre-administration of recombinant H. pylori NAP (rNAP) could prevent ovalbumin (OVA)-induced allergic asthma in mice.. Mice were exposed to purified rNAP through intraperitoneal injection or inhalation and then sensitized with OVA. Following a challenge with aerosolized OVA, the bronchoalveolar lavage fluid (BALF) cell count, lung tissue histology, BALF cytokines and serum IgE were evaluated.. Both intraperitoneal injection and inhalation of rNAP prior to OVA sensitization significantly reduced eosinophil accumulation and inflammatory infiltration in lung tissue in OVA-induced asthma mice; eosinophils were reduced in the BALF of rNAP-treated mice. In addition, IL-4 and IL-13 levels were lower (P < 0.01), IL-10 and IFN-γ levels were higher (P < 0.01) and IgE serum levels were lower (P < 0.01) in the treated groups compared to the control group.. Systemic and mucosal pre-administration of rNAP could suppress the development of OVA-induced asthma in mice; rNAP may be utilized as part of novel strategies for the prevention or treatment of allergic diseases.

Topics: Administration, Inhalation; Animals; Asthma; Bacterial Proteins; Bronchoalveolar Lavage Fluid; Child; Cytokines; Female; Humans; Hypersensitivity; Immunologic Factors; Injections, Intraperitoneal; Leukocyte Count; Lung; Male; Mice, Inbred BALB C; Ovalbumin; Recombinant Proteins; Treatment Outcome

5-Hydroxymethylfurfural (5-HMF) is one of the most important products of the Maillard reaction. In recent years, many profitable biological effects of this compound have been demonstrated. This study sought to elucidate the anti-allergic effect of 5-HMF by investigating some selected components of the immune response in BALB/c mice immunised with ovalbumin (OVA).. Immunised animals had an increased level of serum total and OVA-specific antibodies when compared to the control (P < 0.01).We found that the OVA-induced increase in serum IgE and OVA-specific IgE were significantly suppressed in the groups treated with 5-HMF (P < 0.05). Moreover, interleukin-4 (IL-4) and interferon gamma (IFN-γ) were significantly reduced in a dose-independent manner when compared to the sensitised group (P < 0.05).. 5-HMF inhibited the up-regulation of total and OVA-specific IgE through the suppression of the Th2-type immune response in immunised BALB/c mice. 5-HMF could therefore be a novel therapeutic approach for the prevention of IgE-mediated allergic diseases. © 2017 Society of Chemical Industry.

Topics: Animals; Anti-Allergic Agents; Female; Furaldehyde; Humans; Hypersensitivity; Immunity, Humoral; Immunoglobulin E; Interferon-gamma; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells

This study investigated allergy immunotherapy potential of Lactobacillus paracasei L9 to prevent or mitigate the particulate matter 2.5 (PM2.5) enhanced pre-existing asthma in mice. Firstly, we used a mouse model of asthma (a 21-day ovalbumin (OVA) sensitization and challenge model) followed by PM2.5 exposure twice on the same day of the last challenge. PM2.5 was collected from the urban area of Beijing and underwent analysis for metals and polycyclic aromatic hydrocarbon contents. The results showed that PM2.5 exposure enhanced airway hyper-responsiveness (AHR) and lead to a mixed Th2/ IL-17 response in asthmatic mice. Secondly, the PM2.5 exposed asthmatic mice were orally administered with L9 (4×107, 4×109 CFU/mouse, day) from the day of first sensitization to the endpoint, for 20 days, to investigate the potential mitigative effect of L9 on asthma. The results showed that L9 ameliorated PM2.5 exposure enhanced AHR with an approximate 50% decrease in total airway resistance response to methacholine (48 mg/ml). L9 also prevented the exacerbated eosinophil and neutrophil infiltration in bronchoalveolar lavage fluid (BALF), and decreased the serum level of total IgE and OVA-specific IgG1 by 0.44-fold and 0.3-fold, respectively. Additionally, cytokine production showed that L9 significantly decreased T-helper cell type 2 (Th2)-related cytokines (IL-4, -5, -13) and elevated levels of Th1 related IFN-γ in BALF. L9 also reduced the level of IL-17A and increased the level of TGF-β. Taken together, these results indicate that L9 may exert the anti-allergic benefit, possibly through rebalancing Th1/Th2 immune response and modulating IL-17 pro-inflammatory immune response. Thus, L9 is a promising candidate for preventing PM exposure enhanced pre-existing asthma.

Topics: Administration, Oral; Animals; Asthma; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunotherapy; Lacticaseibacillus paracasei; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Particulate Matter; Probiotics; Th1 Cells; Th2 Cells

The small GTPase ras homolog enriched in brain (Rheb) is a downstream target of tuberous sclerosis complex 1/2 (TSC1/2) and an upstream activator of the mechanistic target of rapamycin complex 1 (mTORC1), the emerging essential modulator of M1/M2 balance in macrophages. However, the role and regulatory mechanisms of Rheb in macrophage polarization and allergic asthma are not known. In the present study, we utilized a mouse model with myeloid cell-specific deletion of the Rheb1 gene and an ovalbumin (OVA)-induced allergic asthma model to investigate the role of Rheb1 in allergic asthma and macrophage polarization. Increased activity of Rheb1 and mTORC1 was observed in myeloid cells of C57BL/6 mice with OVA-induced asthma. In an OVA-induced asthma model, Rheb1-KO mice demonstrated a more serious inflammatory response, more mucus production, enhanced airway hyper-responsiveness, and greater eosinophil numbers in bronchoalveolar lavage fluid (BALF). They also showed increased numbers of bone marrow macrophages and BALF myeloid cells, elevated M2 polarization and reduced M1 polarization of macrophages. Thus, we have established that Rheb1 is critical for the polarization of macrophages and inhibition of allergic asthma. Deletion of Rheb1 enhances M2 polarization but decreases M1 polarization in alveolar macrophages, leading to the aggravation of OVA-induced allergic asthma.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Gene Deletion; Hypersensitivity; Macrophage Activation; Macrophages; Mechanistic Target of Rapamycin Complex 1; Mice; Mice, Inbred C57BL; Myeloid Cells; Ovalbumin; Protein Binding; Ras Homolog Enriched in Brain Protein; Signal Transduction; Th1 Cells; Th2 Cells

The association between lead exposure and respiratory diseases including asthma is controversial. Some studies indicate that exposure to environmental lead pollution may cause asthma; however, there is not sufficient data in this regard. The effect of lead on lung pathological findings and serum inflammatory mediators in sensitized and non-sensitized guinea pigs exposed to inhaled lead was examined. Eleven animal groups including control, sensitized, three groups of non sensitized animals, three groups during sensitization, and three groups after sensitization exposed to aerosol of three lead concentrations (n = 6 for each group) were studied. Serum inflammatory mediators levels and lung pathological changes were evaluated. All pathological changes and serum ET-1, EPO, NO levels were significantly higher in the sensitized and non sensitized animals exposed to lead than control group (p < 0.05 to p < 0.001). There was no significant difference between non sensitized groups exposed to high lead concentration and sensitized group. Serum inflammatory mediators levels and pathological findings in sensitized groups exposed to lead both during and after sensitization were significantly higher than sensitized non exposed group (p < 0.05 to p < 0.001). The data of exposed animals to high lead concentration were significantly higher than those of medium and low concentrations; those of medium concentration were also higher than low concentration (p < 0.05 to p < 0.001). In summary, the present study indicates that exposure to inhaled lead is able to induce respiratory changes similar to asthma. In addition, the results indicated that exposure to environmental lead is able to aggravate asthma severity both during development of asthma or after its manifestation.

Topics: Administration, Inhalation; Animals; Endothelin-1; Eosinophil Peroxidase; Female; Guinea Pigs; Hypersensitivity; Inflammation; Lead; Lung; Male; Nitric Oxide; Ovalbumin

The disruption of epithelial barrier integrity is an important factor in the pathogenesis of various immune disorders. However, the restitution of the compromised barrier functions is difficult. This study investigates the regulation of TWIK-related potassium channel-1 (Trek1) in the restitution of intestinal epithelial barrier functions. The human colon epithelial cell line T84 was cultured in monolayers and used to observe epithelial barrier functions in vitro. An intestinal allergy mouse model was created. Cytokine levels were determined by enzyme-linked immunosorbent assay and western blotting. The results showed that Trek1 deficiency induced T84 monolayer barrier disruption. Allergic responses markedly suppressed the expression of Trek1 in the intestinal epithelia via activating the mitogen-activated protein kinase pathways and increasing the expression of histone deacetylase-1. The inhibition of histone deacetylase-1 by sodium butyrate or the administration of a butyrate-producing probiotic (Clostridium butyricum) restored the intestinal epithelial barrier functions and markedly enhanced the effect of antigen-specific immunotherapy. The data suggest that Trek1 is required for the maintenance of intestinal epithelial barrier integrity. Allergic responses induce an insufficiency of Trek1 expression in the intestinal epithelia. Trek1 expression facilitates the restoration of intestinal epithelial barrier functions in an allergic environment.

Topics: Animals; Butyric Acid; Cell Line; Clostridium butyricum; Cytokines; Eosinophils; Epithelial Cells; Gene Expression Regulation; Histone Deacetylase 1; Histone Deacetylase Inhibitors; Humans; Hydroxamic Acids; Hypersensitivity; Intestinal Mucosa; Mast Cells; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Ovalbumin; Potassium Channels, Tandem Pore Domain; Probiotics; Signal Transduction

The transcription factor promyelocytic leukemia zinc finger (PLZF) is transiently expressed during development of type 2 innate lymphoid cells (ILC2s) but is not present at the mature stage. We hypothesized that PLZF-deficient ILC2s have functional defects in the innate allergic response and represent a tool for studying innate immunity in a mouse with a functional adaptive immune response.. We determined the consequences of PLZF deficiency on ILC2 function in response to innate and adaptive immune stimuli by using PLZF(-/-) mice and mixed wild-type:PLZF(-/-) bone marrow chimeras.. PLZF(-/-) mice, wild-type littermates, or mixed bone marrow chimeras were treated with the protease allergen papain or the cytokines IL-25 and IL-33 or infected with the helminth Nippostrongylus brasiliensis to induce innate type 2 allergic responses. Mice were sensitized with intraperitoneal ovalbumin-alum, followed by intranasal challenge with ovalbumin alone, to induce adaptive TH2 responses. Lungs were analyzed for immune cell subsets, and alveolar lavage fluid was analyzed for ILC2-derived cytokines. In addition, ILC2s were stimulated ex vivo for their capacity to release type 2 cytokines.. PLZF-deficient lung ILC2s exhibit a cell-intrinsic defect in the secretion of IL-5 and IL-13 in response to innate stimuli, resulting in defective recruitment of eosinophils and goblet cell hyperplasia. In contrast, the adaptive allergic inflammatory response to ovalbumin and alum was unimpaired.. PLZF expression at the innate lymphoid cell precursor stage has a long-range effect on the functional properties of mature ILC2s and highlights the importance of these cells for innate allergic responses in otherwise immunocompetent mice.

Topics: Adaptive Immunity; Adoptive Transfer; Allergens; Animals; Antigens, Surface; Biomarkers; Bone Marrow Transplantation; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Helminthiasis; Helminths; Hypersensitivity; Immunity, Innate; Immunophenotyping; Interleukin-33; Interleukins; Kruppel-Like Transcription Factors; Lymphocyte Activation; Lymphocyte Subsets; Mice; Mice, Knockout; Ovalbumin; Papain; Promyelocytic Leukemia Zinc Finger Protein; Pulmonary Eosinophilia; Th2 Cells

P2X4 receptor (P2X4R) is the most widely expressed subtype of the P2XRs in the purinergic receptor family. Adenosine triphosphate (ATP), a ligand for this receptor, has been implicated in the pathogenesis of asthma. ATP‑P2X4R signaling is involved in pulmonary vascular remodeling, and in the proliferation and differentiation of airway and alveolar epithelial cell lines. However, the role of P2X4R in asthma remains to be elucidated. This aim of the present study was to investigate the effects of P2X4R in a murine experimental asthma model. The asthmatic model was established by the inhalation of ovalbumin (OVA) in BALB/c mice. The mice were treated with P2X4R‑specific agonists and antagonists to investigate the role of this receptor in vivo. Pathological changes in the bronchi and lung tissues were examined using hematoxylin and eosin staining, Masson's trichrome staining and Alcian blue staining. The inflammatory cells in the bronchoalveolar lavage fluid were counted, and the expression levels of P2X4R, α‑smooth muscle actin (α‑SMA) and proliferating cell nuclear antigen (PCNA) were detected using western blotting. In the OVA‑challenged mice, inflammation, infiltration, collagen deposition, mucus production, and the expression levels of P2X4R and PCNA were all increased; however, the expression of α‑SMA was decreased, compared with the mice in the control group. Whereas treatment with the P2X4R agonist, ATP, enhanced the allergic reaction, treatment with the P2X4R antagonist, 5‑BDBD, attenuated the allergic reaction. The results suggested that ATP‑P2X4R signaling may not only contribute to airway inflammation, but it may also contribute to airway remodeling in allergic asthma in mice.

Topics: Actins; Airway Remodeling; Animals; Bronchi; Bronchoalveolar Lavage Fluid; Cell Count; Collagen; Female; Goblet Cells; Hyperplasia; Hypersensitivity; Inflammation; Lung; Mice, Inbred BALB C; Mucus; Ovalbumin; Proliferating Cell Nuclear Antigen; Receptors, Purinergic P2X4

Asthma is a disease characterized by chronic relapsing airways, and its etiology remains incompletely understood. To better understand the metabolic phenotypes of asthma, we investigated a plasma metabolic signature associated with allergic asthma in ovalbumin (OVA)-sensitized mice by using ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Sixteen metabolites were characterized as potential pathological biomarkers related to asthma. Among them, 6 (dodecanoic acid (P1), myristic acid (P2), phytosphingosine (P3), sphinganine (P4), inosine (P13) and taurocholic acid (P15)) were first reported to have potential relevance in the pathogenesis of experimental asthma. The identified potential biomarkers were involved in 6 metabolic pathways and achieved the most entire metabolome contributing to the formation of allergic asthma. Purine metabolism was the most prominently influenced in OVA-induced asthma mice according to the metabolic pathway analysis (MetPA), suggesting that significantly changes in inflammatory responses in the pathophysiologic process of asthma. The metabolites of purine metabolism, especially uric acid (P12) and inosine (P13), may denote their potential as targeted biomarkers related to experimental asthma. The decreased plasma uric acid (P12) suggested that inflammation responses of allergic asthma inhibited the activity of xanthine oxidase in purine metabolism, and manifested the severity of asthma exacerbation. The increased level of inosine (P13) suggests that inflammatory cells induce adenosine triphosphate (ATP) breakdown, resulting in excessive expression of adenosine deaminase (ADA) in the formation of allergic asthma. These findings provided a novel perspective on the metabolites signatures related to allergic asthma, which provided us with new insights into the pathogenesis of asthma, and the discovery of targets for clinical diagnosis and treatment.

Topics: Animals; Asthma; Female; Hypersensitivity; Metabolomics; Mice; Mice, Inbred BALB C; Ovalbumin; Plasma; Purines

Nonallergic hypersensitivity reaction (NHR) accounts for more than 77% of all immune-mediated immediate hypersensitivity reactions and has become a serious threat to public health. Here, proteomics was used to study the NHR mechanism of two typical substances, the compound 4880 and ovalbumin. Twelve different proteins were suggested as potential biomarkers for examining the NHR mechanism, and our results revealed that the mechanism mainly encompassed 2 processes, i.e., generation and effect processes. The generation process could be classified as direct stimulation, complement (classical and alternative), coagulation, kallikrein-kinin, and integrated pathways. Thus glutathione peroxidase 1, terminal complement complex (complement factor 4d and Bb), coagulation 13, kininogen-1, and IgE could be used as candidate biomarkers for the indication of the corresponding pathways respectively, the proteins were further confirmed by ELISA. And the effect process was mainly composed of histamine as well as proteins such as DCD and MYLPF, which could be used as important indices for the symptoms of NHR. Our study differs from previous studies in that C4880 was found to not only be involved in the direct stimulation pathway, but also in the activated complement and kallikrein-kinin pathways through the coagulation pathway. We also report for the first time that ovalbumin-induced NHR could be a combination of the coagulation, classical complement, and integrated pathways.

Topics: Animals; Behavior, Animal; Chromatography, Liquid; Enzyme-Linked Immunosorbent Assay; Histamine; Hypersensitivity; Immunoglobulin E; Male; Ovalbumin; p-Methoxy-N-methylphenethylamine; Peptides; Proteomics; Rats; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization

Allergic eye disease, as in most forms of atopy, ranges in severity among individuals from immediate hypersensitivity to a severe and debilitating chronic disease. Dendritic cells play a key role in stimulating pathogenic T cells in allergen re-exposure, or secondary responses. However, molecular cues by dendritic cells underpinning allergic T cell response levels and the impact that this control has on consequent severity of allergic disease are poorly understood. Here, we show that a deficiency in thrombospondin-1, a matricellular protein known to affect immune function, has subsequent effects on downstream T cell responses during allergy, as revealed in an established mouse model of allergic eye disease. More specifically, we demonstrate that a thrombospondin-1 deficiency specific to dendritic cells leads to heightened secondary T cell responses and consequent clinical disease. Interestingly, whereas thrombospondin-1-deficient dendritic cells augmented activity of allergen-primed T cells, this increase was not recapitulated with naïve T cells in vitro. The role of dendritic cell-derived thrombospondin-1 in regulating secondary allergic T cell responses was confirmed in vivo, as local transfer of thrombospondin-1-sufficient dendritic cells to the ocular mucosa of thrombospondin-1 null hosts prevented the development of augmented secondary T cell responses and heightened allergic eye disease clinical responses. Finally, we demonstrate that topical instillation of thrombospondin-1-derived peptide reduces T cell activity and clinical progression of allergic eye disease. Taken together, this study reveals an important modulatory role of dendritic cell-derived thrombospondin-1 on secondary allergic T cell responses and suggests the possible dysregulation of dendritic cell-derived thrombospondin-1 expression as a factor in allergic eye disease severity.

Topics: Allergens; Animals; Dendritic Cells; Eye Diseases; Hypersensitivity; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; T-Lymphocytes; Thrombospondin 1

Inflammation drives asthma and atherosclerosis. Clinical studies suggest that asthmatic patients have a high risk of atherosclerosis. Yet this hypothesis remains uncertain, given that Th2 imbalance causes asthma whereas Th1 immunity promotes atherosclerosis. In this study, chronic allergic lung inflammation (ALI) was induced in mice by ovalbumin sensitization and challenge. Acute ALI was induced in mice by ovalbumin and aluminum sensitization and ovalbumin challenge. Atherosclerosis was produced in apolipoprotein E-deficient (Apoe(-/-)) mice with a Western diet. When chronic ALI and atherosclerosis were produced simultaneously, ALI increased atherosclerotic lesion size, lesion inflammatory cell content, elastin fragmentation, smooth muscle cell (SMC) loss, lesion cell proliferation, and apoptosis. Production of acute ALI before atherogenesis did not affect lesion size, but increased atherosclerotic lesion CD4(+) T cells, lesion SMC loss, angiogenesis, and apoptosis. Production of acute ALI after atherogenesis also did not change atherosclerotic lesion area, but increased lesion elastin fragmentation, cell proliferation, and apoptosis. In mice with chronic ALI and diet-induced atherosclerosis, daily inhalation of a mast cell inhibitor or corticosteroid significantly reduced atherosclerotic lesion T-cell and mast cell contents, SMC loss, angiogenesis, and cell proliferation and apoptosis, although these drugs did not affect lesion area, compared with those that received vehicle treatment. In conclusion, both chronic and acute ALI promote atherogenesis or aortic lesion pathology, regardless whether ALI occurred before, after, or at the same time as atherogenesis. Antiasthmatic medication can efficiently mitigate atherosclerotic lesion pathology.

Topics: Animals; Apolipoproteins E; Atherosclerosis; Budesonide; Chronic Disease; Disease Progression; Glucocorticoids; Hypersensitivity; Inflammation; Ketotifen; Male; Mast Cells; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pneumonia

To evaluate the influence of lactation on lung immune function during allergic inflammation.. Female rats, 60-90days old, were divided into three groups: no lung allergy virgins (N group), ovalbumin (OVA)-immunized and sensitized virgins (V group), and OVA-immunized and sensitized lactating females (L group). On gestation day (GD) 10, all animals in L group received a subcutaneous injection of 0.1mg·kg(-1) OVA plus aluminum hydroxide. On GD17, the L group received a subcutaneous booster injection of 10μg OVA plus 10mg aluminum hydroxide. After 7days, an inhalatory challenge with 1% OVA was given in 15min sessions for 3 consecutive days. Animals from the V group received the same treatment, meaning both tests and time intervals between OVA treatment and inhalatory challenge were the same as in the L group. Twenty-four hours after the last inhalation session, the animals were euthanized, and the following tests were performed: total and differential bronchoalveolar lavage (BAL) and femoral marrow lavage (FML) leukocyte counts, quantification of tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ) levels in BAL fluid, and quantification of plasma corticosterone and catecholamine levels.. The L group presented lower BAL total leukocyte counts and decreases in the number of eosinophils and macrophages compared with the V group. They also expressed higher BAL IFN-γ and lower plasma corticosterone levels. Plasma norepinephrine levels were higher in the L group than in the N and V groups.. Lactating female rats presented less intense allergic lung inflammation. Our findings suggest that lactation may protect females from asthmatic crises.

Topics: Administration, Inhalation; Aluminum Hydroxide; Animals; Bone Marrow; Bronchoalveolar Lavage Fluid; Catecholamines; Corticosterone; Female; Hypersensitivity; Inflammation; Interferon-gamma; Lactation; Leukocyte Count; Lung; Ovalbumin; Rats; Tumor Necrosis Factor-alpha

We investigated the effects of pan-class I PI3K inhibitor, ZSTK474 on vascular remodeling using a murine model of allergic vasculitis with eosinophil infiltration.. C57BL/6 mice were sensitized with OVA. The positive controls were exposed to aerosolized OVA daily for 7 days. The other group of mice were administered ZSTK474 (30 mg/kg, p.o. daily) in parallel with daily exposure to aerosolized OVA for 7 days. On the 3rd and 7th day, bronchoalveolar lavage (BAL) was performed and the lungs were excised for pathological analysis. Cell differentials were determined and the concentrations of IL-4, IL-5, IL-13 and TGF-βin BAL fluid were measured.. The total cell numbers and eosinophil numbers in BALF were greatly reduced in the ZSTK474-treated group on the 3rd and 7th day after exposure to OVA. The numbers of total white blood cells and eosinophils in the peripheral blood were significantly reduced in the ZSTK474-treated group on the 3rd and 7th day after exposure to OVA. The concentrations of IL-4, IL-5, and IL-13 in BAL fluids were also reduced significantly on the 3rd day in the ZSTK474-treated group. The concentrations of TGF-β in BAL fluids were also reduced significantly on the 3rd and 7th day in the ZSTK474-treated group. The pathological scores reduced significantly in the ZSTK474-treated group compared to the control group.. The PI3K inhibitor, ZSTK474 suppressed pulmonary vascular remodeling in the murine model of allergic vasculitis with eosinophil infiltration. PI3K signal transduction may have a critical role in the immunological process that induces allergic vasculitis.

Topics: Animals; Asthma; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Inflammation; Interleukin-13; Interleukin-4; Interleukin-5; Leukocyte Count; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Phosphoinositide-3 Kinase Inhibitors; Transforming Growth Factor beta; Triazines; Vascular Remodeling; Vasculitis

Allergen sources such as mites, insects, fungi, and pollen contain proteases. Airway exposure to proteases induces allergic airway inflammation and IgE/IgG1 responses via IL-33-dependent mechanisms in mice. We examined the epicutaneous sensitization of mice to a model protease allergen, papain; the effects of tape stripping, which induces epidermal barrier dysfunction; and the atopic march upon a subsequent airway challenge. Papain painting on ear skin and tape stripping cooperatively promoted dermatitis, the skin gene expression of proinflammatory cytokines and growth factors, up-regulation of serum total IgE, and papain-specific IgE/IgG1 induction. Epicutaneous sensitization induced T helper (Th) 2 cells and Th17 differentiation in draining lymph nodes. Ovalbumin and protease inhibitor-treated papain induced no or weak responses, whereas the co-administration of ovalbumin and papain promoted ovalbumin-specific IgE/IgG1 induction. Wild-type and IL-33-deficient mice showed similar responses in the epicutaneous sensitization phase. The subsequent airway papain challenge induced airway eosinophilia and maintained high papain-specific IgE levels in an IL-33-dependent manner. These results suggest that allergen source-derived protease activity and mechanical barrier damage such as that caused by scratching cooperatively promote epicutaneous sensitization and skin inflammation and that IL-33 is dispensable for epicutaneous sensitization but is crucial in the atopic march upon a subsequent airway low-dose encounter with protease allergens.

Topics: Allergens; Animals; Cell Differentiation; Cytokines; Dermatitis; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-33; Mice; Mice, Inbred C57BL; Ovalbumin; Papain; Protease Inhibitors; Real-Time Polymerase Chain Reaction; Skin; Stress, Mechanical; Th17 Cells; Th2 Cells; Wounds and Injuries

Use of the reverse targeting drug delivery system (RT-DDS) is a new targeting strategy based on the specific delivery of drugs to immune cells in antigen-sensitized animals by using antigen-modified liposomes, and it is expected to be a curative treatment for allergic diseases.. Herein, we prepared ovalbumin (OVA)-modified liposomes encapsulating the immunosuppressive drug FK506 (OVA-LipFK) and aimed to demonstrate the delivery selectivity of the liposomes to splenic B cells, and its antiallergic effect in an OVA-sensitized allergic model mouse.. Fluorescently labeled OVA-LipFK was intravenously injected into OVA-sensitized mice, and the intrasplenic localization of liposomes was observed. The antiallergic effect of OVA-LipFK in OVA-sensitized mice was examined by measuring the blood levels of OVA-specific IgE and IgG antibodies.. OVA-LipFK was co-localized to not only B cells but also germinal centers, in the spleen of OVA-sensitized mice. However, there was no accumulation of unmodified liposomes encapsulating FK506 (LipFK) in the splenic B-cell area. In a therapeutic study, OVA-LipFK significantly suppressed the production of both OVA-specific IgE and IgG antibodies in OVA-sensitized mice after the animals had been boosted with OVA, whereas LipFK showed little antiallergic effect.. The present study suggested that the introduction of RT-DDS for use with immunosuppressive drugs could be useful for the treatment of allergic diseases.

Topics: Animals; B-Lymphocytes; Hypersensitivity; Immunosuppressive Agents; Liposomes; Mice; Ovalbumin; Spleen; Tacrolimus; Tissue Distribution

Uvaol, a triterpene present in olives and virgin olive oil, has been shown to possess anti-inflammatory properties and antioxidant effects. However, until now, no studies have demonstrated its potential effects on allergic inflammation. The aim of this study was to evaluate the anti-inflammatory effects of uvaol in a mouse model of allergy characterized by eosinophil-dominant inflammation in actively sensitized mice. The anti-inflammatory effect of uvaol was analyzed in two murine models of allergic inflammation (pleurisy and asthma). In these models, Swiss mice were sensitized and challenged with ovalbumin (OVA). In the pleurisy model, the pleural eosinophilic inflammation and IL-5 concentrations were examined 24h after the OVA challenge, while in the asthma model were examined the airway inflammation via bronchoalveolar lavage (BAL) fluid cytology and lung histopathology analyses. Our results showed that uvaol decreased the accumulation of eosinophils and the concentration of IL-5 in pleural effluent. Uvaol also demonstrated important anti-inflammatory activity by inhibiting production of IL-5 and influx of leukocytes, mainly of eosinophils, in BAL fluid, but without interfering with levels of reactive oxygen species in leukocytes. Moreover, the eosinophil infiltration, mucus production, number of alveoli that collapsed, and IL-5 levels in the lung were clearly decreased by uvaol treatment. These findings indicate that uvaol can be a good candidate for the treatment of allergic inflammation by inhibiting eosinophil influx and IL-5 production in ovalbumin-induced allergy.

Topics: Allergens; Animals; Eosinophils; Hypersensitivity; Inflammation; Lung; Male; Mice; Ovalbumin; Pleurisy; Triterpenes

Titanium dioxide nanoparticles (nTiO2) previously considered to possess relatively low toxicity both in vitro and in vivo, although classified as possibly carcinogenic to humans. Also, their adjuvant potential has been reported to promote allergic sensitization and modulate immune responses. Previously, in OVA induced mouse model of asthma we found high expression of Socs3 and low expression of Stat3 and IL-6. However, a clear understanding regarding the signaling pathways associated with nTiO2 adjuvant effect in mouse model of asthma is lacking. In the present study we investigated the status of Stat3/IL-6 and Socs3 and their relationship with NF-κB, with nTiO2 as an adjuvant in mouse model of asthma. nTiO2 when administered with ovalbumin (OVA) during sensitization phase augmented airway hyper-responsiveness (AHR), biochemical markers of lung damage and a mixed Th2/Th1 dependent immune response. At the same time, we observed significant elevation in the levels of Stat3, Socs3, NF-κB, IL-6 and TNF-α. Furthermore, transient in vivo blocking of NF-κB by NF-κB p65 siRNA, downregulated the expression of Socs3, IL-6 and TNF-α. Our study, thus, shows that nTiO2 exacerbate the inflammatory responses in lungs of pre-sensitized allergic individuals and that these changes are regulated via NF-κB pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Gene Knockdown Techniques; Hypersensitivity; Inflammation; Lung; Mice, Inbred BALB C; Models, Biological; Nanoparticles; NF-kappa B; Ovalbumin; Signal Transduction; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein; Th1 Cells; Th2 Cells; Titanium; Up-Regulation

Few efficacious topical therapies exist for chronic rhinosinusitis (CRS). The lack of a reproducible mouse model of CRS limits the pilot testing of potential novel anti-inflammatory therapies. Although the ovalbumin-induced mouse model of sinonasal inflammation is commonly used, it is difficult to reproduce and can generate variable histologic results. In this study, we explore a variation of this model in different strains of mice and explore various inflammatory cytokines as reproducible molecular markers of inflammation.. Allergic sinonasal inflammation was generated in BALB/c and C57BL/6 mice using intraperitoneal high-dose injections of ovalbumin (Ova; Sigma Chemical Co.) followed by 10 days of high-dose intranasal sensitization. Real-time polymerase chain reaction (RT-PCR) for eotaxin, interleukin 4 (IL-4), and IL-13 were measured from sinonasal mucosa. We also pilot tested a known topical budesonide to characterize the anti-inflammatory response. Histological sections were analyzed for epithelial thickness and eosinophilia.. Both BALB/c and C57BL/6 mice consistently showed increases in T helper 2 (Th2) cytokines after sensitization with high-dose Ova (p < 0.0001) when compared to controls. There were also significant increases in epithelial thickening in Ova-sensitized mice and eosinophilia in both BALB/c and C57BL/6 strains. In addition, topical budesonide significantly reduced anti-inflammatory cytokines, eosinophilia, and epithelial thickness.. Our variation of the ovalbumin-induced mouse model of sinonasal inflammation in both BALB/c and C57BL/6 mice provides an efficacious model for testing potential topical anti-inflammatory therapies for CRS. The utilization of sinonasal mucosal Th2 cytokines along with histologic markers provides a consistent and quantifiable marker of inflammation in assessing the efficacy of candidate drugs.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Budesonide; Cytokines; Disease Models, Animal; Eosinophilia; Female; Hypersensitivity; Male; Mice, Inbred BALB C; Mice, Inbred C57BL; Nasal Lavage Fluid; Nasal Mucosa; Ovalbumin; RNA, Messenger; Sinusitis

The present study describes the distribution of CD4+CD8+ double-positive (DP) T cells in various immune compartments of mice with ovalbumin (OVA)-induced allergic asthma. It was found that the absolute number of DP T cells was considerably increased in the mediastinal lymph nodes and lungs of asthmatic mice as compared with that determined in the healthy subjects. On the contrary, the absolute counts of DP T cells was significantly decreased in the head and neck lymph nodes, and in peripheral blood of OVA-immunized mice. These results suggest that DP T cells may be involved in the pathogenesis of allergic asthma.

Topics: Animals; Asthma; Hypersensitivity; Lung; Lymph Nodes; Mice; Ovalbumin; T-Lymphocyte Subsets

Expression of voltage-gated sodium channels (Nav) takes place in the airways and the role of Nav1.7 and Nav1.8 in the control of airway's defense reflexes has been confirmed. The activation of Nav channels is crucial for cough initiation and airway smooth muscle reactivity, but it is unknown whether these channels regulate ciliary beating. This study evaluated the involvement of Nav1.7 and Nav1.8 channels in the airway defense mechanisms using their pharmacological blockers in healthy guinea pigs and in the experimental allergic asthma model. Asthma was modeled by ovalbumin sensitization over a period of 21 days. Blockade of Nav1.7 channels significantly decreased airway smooth muscle reactivity in vivo, the number of cough efforts, and the cilia beat frequency in healthy animals. In the allergic asthma model, blockade of Nav1.8 efficiently relieved symptoms of asthma, without adversely affecting cilia beat frequency. The study demonstrates that Nav1.8 channel antagonism has a potential to alleviate cough and bronchial hyperreactivity in asthma.

Topics: Animals; Asthma; Cilia; Cough; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Male; Muscle, Smooth; Ovalbumin; Respiratory Mucosa; Sodium Channel Blockers; Voltage-Gated Sodium Channels

Oxidative stress plays a pivotal role in the pathogenesis of asthma. Aquaporin-3 (AQP3) is a small transmembrane water/glycerol channel that may facilitate the membrane uptake of hydrogen peroxide (H2O2). Here we report that AQP3 potentiates ovalbumin (OVA)-induced murine asthma by mediating both chemokine production from alveolar macrophages and T cell trafficking. AQP3 deficient (AQP3(-/-)) mice exhibited significantly reduced airway inflammation compared to wild-type mice. Adoptive transfer experiments showed reduced airway eosinophilic inflammation in mice receiving OVA-sensitized splenocytes from AQP3(-/-) mice compared with wild-type mice after OVA challenge, consistently with fewer CD4(+) T cells from AQP3(-/-) mice migrating to the lung than from wild-type mice. Additionally, in vivo and vitro experiments indicated that AQP3 induced the production of some chemokines such as CCL24 and CCL22 through regulating the amount of cellular H2O2 in M2 polarized alveolar macrophages. These results imply a critical role of AQP3 in asthma, and AQP3 may be a novel therapeutic target.

Topics: Animals; Aquaporin 3; Asthma; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cell Count; Cell Membrane Permeability; Chemokines; Gene Expression Regulation; Hydrogen Peroxide; Hypersensitivity; Lymph Nodes; Macrophages, Alveolar; Mice, Inbred C57BL; Ovalbumin; Pneumonia; RNA, Messenger; Spleen

This study was conducted to evaluate the potent immunomodulatory effects of Lactobacilli and their possible anti-allergic effects.. Lactobacillus plantarum LP45 (LP45), Lactobacillus acidophilus La28 (La28), Lactobacillus acidophilus 6091 (6091), Lactobacillus rhamnosus GG (LGG) were orally administrated to male BALB/C mice, respectively for 28 d. The immune organ index, serum Th1 cytokines [interferon-γ (IFN-γ), interleukin-12 (IL-12)] and Th2 cytokines IL-6 of the tested mice were analyzed with ELISA after intervention. Furthermore, La28, 6901 were also orally fed to ovalbumin (OVA)-sensitized male BALB/C. The serum total IgE of the tested mice was analyzed with ELISA after intervention.. No statistical difference was found in immune organ index among the tested four strains. La28 significantly decreased serum IL-6 of the tested mice after 14 d and 28 d compared to those in control (P < 0.05). After 28 d, 6091 also significantly reduced serum IL-6 of the tested mice (P < 0.05). La28 significantly suppressed the increase of serum total IgE of the tested mice (P < 0.05).. The present study indicates that the immunomodulatory effects of Lactobacilli might be strain-dependent. Among the tested strains of Lactobacilli, La28 and 6091 may have possibility to influence the Th2 immunity of host animal. La28 may also posse potent ability to alter IgE mediated allergy by the way to affect Th1/Th2 balance of host animal.

Topics: Animals; Hypersensitivity; Immunoglobulin E; Immunomodulation; Interferon-gamma; Interleukin-12; Interleukin-6; Lactobacillus; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th1-Th2 Balance

Using an experimental model of allergic asthma, we evaluated the anti-asthmatic potential of polyphenol flavonol derivate morin after either acute or long-term treatment of male OVA-sensitised guinea pigs.. The following methods were used in experiments: the in-vitro tracheal smooth muscle contraction induced by histamine; the changes in specific airway resistance (sRaw) to histamine and the sensitivity of a chemically induced cough reflex both via an in-vivo method; the serum and BALF concentrations' analysis of the inflammatory cytokines interleukin IL-4, IL-5, IL-13; and lung tissue infiltration by eosinophils and mastocytes.. Our data show that acute morin (30 mg/kg) and chronic 21-day morin (30 mg/kg/day) administration had a comparable antitussive efficiency with opioid antitussive codeine. Acute morin bronchodilatory activity defined by in-vivo sRaw decline did not reach SABA salbutamol effect. However, bronchodilatory efficiency of morin after long-term administration was by 34% higher as effect of LABA salmeterol. The 21-day morin treatment of OVA-sensitised guinea pigs reduced the serum, BALF levels of IL-4 and IL-13, lung tissue eosinophil and mastocyte infiltration comparable with corticosteroid budesonide.. In summary, morin represents very rational target for additional studies as potential substance for control as well as prevention of asthma inflammation and symptoms.

Topics: Airway Resistance; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antitussive Agents; Asthma; Bronchoalveolar Lavage Fluid; Bronchodilator Agents; Cough; Cytokines; Eosinophils; Flavonoids; Guinea Pigs; Histamine; Hypersensitivity; Inflammation; Lung; Male; Ovalbumin; Phytotherapy; Plant Extracts; Trachea

Natural products are considered as an essential source for the search of new drugs. Pistacia integerrima galls (PI) have been used for the treatment of asthma and cough in traditional system of medicine.. Current study investigates the immunomodulatory and anti-inflammatory activities of P. integerrima in mouse model of ovalbumin-induced allergic asthma.. Mice were intraperitoneally sensitized and subsequently challenged intranasally with ovalbumin to induce allergic asthma. Experimental group mice were treated with methanol extract of P. integerrima extract (200mg/kg b. w.) and Methylprednisolone (MP) (15mg/kg b. w.) for 07 consecutive days, alongside intranasal challenge. Lung tissues were stained with Hematoxyline and Eosin (H & E), and Periodic Acid-Schiff (PAS) stains for histopathological evaluation. Lung wet/dry weight ratio was measured as an index of lung tissue edema. Albumin was injected in the right ear 24h before sacrificing the mice and difference of weight was taken as a degree of delayed type hypersensitivity (DTH). mRNA expression levels of TNF-α, IL-4, IL-5, Aquaporin-1 (AQP1), and AQP5 were evaluated using reverse transcription polymerase chain reaction (RT-PCR) followed by gel electrophoresis.. The data showed both PI extract and MP significantly alleviated DTH and nearly normalized total leukocyte count and differential leukocyte count in both blood and BALF. We found significantly suppressed goblet cell hyperplasia and inflammatory cell infiltration after treatment with both PI extract and MP. Expression levels of TNF-α, IL-4, and IL-5 were also found significantly reduced after treatment with both PI extract and MP, which might have resulted in the amelioration of airway inflammation. Current study displayed that both PI extract and MP significantly decreased lung wet/dry ratio, suggesting reduction in pulmonary edema. RT-PCR analysis showed significant increase in AQP1 and AQP5 expression levels after treatment with both PI extract and MP, which might have caused the alleviation of pulmonary edema.. Our study displays that P. integerrima possesses significant anti-asthmatic activity which may be attributed to reduction in TNF-α, IL-4, and IL-5 expression levels, and increase in AQP1 and AQP5 expression levels.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Aquaporin 1; Aquaporin 5; Asthma; Female; Hypersensitivity; Inflammation; Interleukin-4; Interleukin-5; Methylprednisolone; Mice; Mice, Inbred BALB C; Ovalbumin; Pistacia; Plant Extracts; Pulmonary Edema; Tumor Necrosis Factor-alpha

Overproduction of mucus is a hallmark of asthma. The aim of this study was to identify potentially effective therapies for removing excess mucus. The role of voltage-gated (Kir 6.1, KCa 1.1) and store-operated ion channels (SOC, CRAC) in respiratory cilia, relating to the tracheal ciliary beat frequency (CBF), was compared under the physiological and allergic airway conditions. Ex vivo experiments were designed to test the local effects of Kir 6.1, KCa 1.1 and CRAC ion channel modulators in a concentration-dependent manner on the CBF. Cilia, obtained with the brushing method, were monitored by a high-speed video camera and analyzed with ciliary analysis software. In natural conditions, a Kir 6.1 opener accelerated CBF, while CRAC blocker slowed it in a concentration-dependent manner. In allergic inflammation, the effect of Kir 6.1 opener was insignificant, with a tendency to decrease CBF. A cilio-inhibitory effect of a CRAC blocker, while gently reduced by allergic inflammation, remained significant. A KCa 1.1 opener turned out to significantly enhance the CBF under the allergic OVA-sensitized conditions. We conclude that optimally attuned concentration of KCa 1.1 openers or special types of bimodal SOC channel blockers, potentially given by inhalation, might benefit asthma.

Topics: Animals; Asthma; Calcium Channel Blockers; Cilia; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Inflammation; Ion Channels; Male; Ovalbumin; Respiratory Mucosa; Trachea

Topics: Acetylation; Animals; Asthma; GATA3 Transcription Factor; Hypersensitivity; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Sirtuins; Th2 Cells

The catalytical isoforms p110γ and p110δ of phosphatidylinositide 3-kinase γ (PI3Kγ) and PI3Kδ play an important role in the pathogenesis of asthma. Two key elements in allergic asthma are increased levels of eosinophils and IgE. Dual pharmacological inhibition of p110γ and p110δ reduces asthma-associated eosinophilic lung infiltration and ameliorates disease symptoms, whereas the absence of enzymatic activity in p110γKOδD910A mice increases IgE and basal eosinophil counts. This suggests that long-term inhibition of p110γ and p110δ might exacerbate asthma. Here, we analysed mice genetically deficient for both catalytical subunits (p110γ/δ-/-) and determined basal IgE and eosinophil levels and the immune response to ovalbumin-induced asthma. Serum concentrations of IgE, IL-5 and eosinophil numbers were significantly increased in p110γ/δ-/- mice compared to single knock-out and wildtype mice. However, p110γ/δ-/- mice were protected against OVA-induced infiltration of eosinophils, neutrophils, T and B cells into lung tissue and bronchoalveolar space. Moreover, p110γ/δ-/- mice, but not single knock-out mice, showed a reduced bronchial hyperresponsiveness. We conclude that increased levels of eosinophils and IgE in p110γ/δ-/- mice do not abolish the protective effect of p110γ/δ-deficiency against OVA-induced allergic airway inflammation.

Topics: Animals; B-Lymphocytes; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Class I Phosphatidylinositol 3-Kinases; Eosinophilia; Eosinophils; Goblet Cells; Hypersensitivity; Immunoglobulin E; Interleukin-5; Lung; Metaplasia; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Pneumonia; T-Lymphocytes

Allergic airway diseases are immune disorders associated with heightened type 2 immune responses and IL-5 and IL-13 production at the site of inflammation. We have previously reported that cyclooxygenase (COX) inhibition by indomethacin augmented allergic airway inflammation in a STAT6-independent manner. However, the key COX product(s) responsible for restraining indomethacin-mediated STAT6-independent allergic inflammation is unknown. In this study, using the mouse model of OVA-induced allergic airway inflammation, we identified that PGI2 receptor (IP) signaling was critical for indomethacin-induced, STAT6-independent proallergic effects. We demonstrated that IP deficiency increased inflammatory cell infiltration, eosinophilia, and IL-5 and IL-13 expression in the lung in a STAT6-independent manner. The augmented STAT6-independent allergic inflammation correlated with enhanced primary immune responses to allergic sensitization and elevated production of multiple inflammatory chemokines (CCL11, CCL17, CCL22, and CXCL12) in the lung after allergen challenge. We also showed that the PGI2 analogue cicaprost inhibited CD4 T cell proliferation and IL-5 and IL-13 expression in vitro, and IP deficiency diminished the stimulatory effect of indomethacin on STAT6-independent IL-5 and IL-13 responses in vivo. The inhibitory effects of PGI2 and the IP signaling pathway on CD4 T cell activation, inflammatory chemokine production, and allergic sensitization and airway inflammation suggest that PGI2 and its analogue iloprost, both Food and Drug Administration-approved drugs, may be useful in treating allergic diseases and asthma. In addition, inhibiting PGI2 signaling by drugs that either block PGI2 production or restrain IP signaling may augment STAT6-independent pathways of allergic inflammation.

Topics: Allergens; Animals; Antihypertensive Agents; Asthma; CD4-Positive T-Lymphocytes; Cell Proliferation; Chemokines; Epoprostenol; Hypersensitivity; Indomethacin; Inflammation; Interleukin-13; Interleukin-5; Lung; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Receptors, Epoprostenol; Signal Transduction; STAT6 Transcription Factor; Th2 Cells

The prevalence of asthma and allergic diseases has risen in recent decades. The etiology of asthma and allergic diseases has not been entirely elucidated.. In this study, we investigated the possibility that programmed vaccination in China may have a potential role in asthma and allergic diseases.. In this animal model, newborn BALB/c mice were randomly divided into three groups: vaccine plus ovalbumin (OVA), OVA, and control. The mice of vaccine plus OVA only group were inoculated with vaccines by following the National Vaccines Inoculation Program in China. Mice of vaccine plus OVA and OVA only groups were sensitized and challenged with OVA. Airway hyperresponsiveness was assessed by lung function and serum interleukin (IL) 4 and interferon (IFN) γ were measured.. The results of lung function showed that mice of the vaccine plus OVA group exhibited an increase in enhanced pause (Penh) compared with that in the OVA group at methacholine concentrations of 6.25 and 12.5 mg/mL (p < 0.05). Serum IL-4 in the vaccine plus OVA group was higher than that in the OVA group (p < 0.01). The serum IFN-γ level in the OVA group was lower than that in the control group (p < 0.01), and also lower than that in the vaccine plus OVA group (p < 0.05). The ratio of IFN-γ to IL-4 both in the OVA and vaccine plus OVA group was lower than that in the control group (p < 0.01).. Results of our study indicated that programmed vaccination in China may have a potential role in the prevalence of asthma and allergic diseases by inducing T-helper 2 cytokine expression and may be responsible for the increasing prevalence of asthma and allergic diseases in China.

Topics: Animals; Asthma; Hypersensitivity; Interferon-gamma; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Th1 Cells; Th2 Cells; Vaccination

Nitrogen dioxide (NO2) is an environmental air pollutant and endogenously generated oxidant that contributes to the exacerbation of respiratory disease and can function as an adjuvant to allergically sensitize to an innocuous inhaled Ag. Because uric acid has been implicated as a mediator of adjuvant activity, we sought to determine whether uric acid was elevated and participated in a mouse model of NO2-promoted allergic sensitization. We found that uric acid was increased in the airways of mice exposed to NO2 and that administration of uricase inhibited the development of OVA-driven allergic airway disease subsequent to OVA challenge, as well as the generation of OVA-specific Abs. However, uricase was itself immunogenic, inducing a uricase-specific adaptive immune response that occurred even when the enzymatic activity of uricase had been inactivated. Inhibition of the OVA-specific response was not due to the capacity of uricase to inhibit the early steps of OVA uptake or processing and presentation by dendritic cells, but occurred at a later step that blocked OVA-specific CD4(+) T cell proliferation and cytokine production. Although blocking uric acid formation by allopurinol did not affect outcomes, administration of ultra-clean human serum albumin at protein concentrations equivalent to that of uricase inhibited NO2-promoted allergic airway disease. These results indicate that, although uric acid levels are elevated in the airways of NO2-exposed mice, the powerful inhibitory effect of uricase administration on allergic sensitization is mediated more through Ag-specific immune deviation than via suppression of allergic sensitization, a mechanism to be considered in the interpretation of results from other experimental systems.

Topics: Adaptive Immunity; Allergens; Allopurinol; Animals; Antigen Presentation; Asthma; Cytokines; Disease Models, Animal; Humans; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Nitrogen Dioxide; Ovalbumin; Serum Albumin; Th2 Cells; Urate Oxidase; Uric Acid

Methylmercury (MeHg) is one of the most toxic environmental pollutants and presents a serious hazard to health worldwide. Although the adverse effects of MeHg, including neurotoxicity, have been studied, its effects on immune function, in particular the immune response, remain unclear. This study examined the effects of low-dose MeHg on immune responses in mice. Mice were orally immunized with ovalbumin (OVA) or subcutaneously injected with mite extract to induce a T-helper 2 (Th2) allergic response. They were then exposed to MeHg (0, 0.02, 1.0, or 5.0 mg·kg(-1)·d(-1)). Immunization with oral OVA or subcutaneous mite extract increased serum levels of OVA-specific immunoglobulin (Ig) E (OVA-IgE), OVA-IgG1, interleukin (IL)-4, and IL-13, and total IgE, total IgG, and IL-13 when compared with levels in non-immunized mice. However, no interferon (IFN)-γ was detected. By contrast, serum levels of OVA-IgE, OVA-IgG1, IL-4, and IL-13, or total IgE, total IgG, and IL-13 in Th2 allergy model mice subsequently treated with MeHg were no higher than those in MeHg-untreated mice. These results suggest that MeHg exposure has no adverse effects on Th2 immune responses in antigen-immunized mice.

Topics: Animals; Brain; Cytokines; Dermatophagoides pteronyssinus; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Methylmercury Compounds; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Mast cell-mediated anaphylactic reactions are involved in many allergic diseases, including asthma and allergic rhinitis. In Korea, where it has been used as a traditional medicine, Rosae Multiflorae fructus (RMF) is known to have potent antioxidative, analgesic, and anti-inflammatory activities and to have no obvious acute toxicity. However, its specific effect on asthma is still unknown. In this study, we evaluated whether or not RMF hot water extracts (RMFW) could inhibit ovalbumin (OVA)-induced allergic asthma and evaluated compound 48/80-induced mast cell activation to elucidate the mechanisms of asthma inhibition by RMFW. Oral administration of RMFW decreased the number of eosinophils and lymphocytes in the lungs of mice challenged by OVA and downregulated histological changes such as eosinophil infiltration, mucus accumulation, goblet cell hyperplasia, and collagen fiber deposits. In addition, RMFW significantly reduced T helper 2 cytokines, TNF-α, IL-4, and IL-6 levels in the BAL fluid of mice challenged by OVA. Moreover, RMFW suppressed compound 48/80-induced rat peritoneal mast cell degranulation and inhibited histamine release from mast cells induced by compound 48/80 in a dose-dependent manner. These results suggest that RMFW may act as an antiallergic agent by inhibitingTh2 cytokine production from Th2 cells and histamine release from mast cells, and could be used as a therapy for patients with Th2-mediated or mast cell-mediated allergic diseases.

Topics: Animals; Anti-Allergic Agents; Asthma; Cytokines; Fruit; Histamine; Histamine Release; Hypersensitivity; Interleukin-4; Interleukin-6; Lung; Male; Mast Cells; Mice, Inbred BALB C; Ovalbumin; p-Methoxy-N-methylphenethylamine; Phytotherapy; Plant Extracts; Rats; Republic of Korea; Rosa; Th2 Cells; Tumor Necrosis Factor-alpha

Extracelluar nucleotides have been identified as regulatory factors in asthmatic pathogenesis by activating purinergic receptors. This research aimed to investigate the function of the purinergic receptor P2Y6 in mediating airway inflammation in allergic asthma. Wild-type (WT) and P2Y6-deficient mice were stimulated with ovalbumin (OVA) to construct asthmatic mouse models. Overexpression of P2Y6 and uridine 5'-diphosphate (UDP)-releasing were demonstrated in lung tissues in ovalbumin-induced asthmatic mice. The release of the cytokine IL-4, mast cell invasion, and the airway remodeling phenotypes were more severe following the application of UDP in asthmatic mice. However, P2Y6 deficiency reduced these asthmatic pathogeneticsymptoms markedly in a mouse model. In vitro, we found that P2Y6 in purified mast cells enhanced the functions of mast cells in the inflammatory response in the asthmatic process by triggering their capability for migration, cytokine secretion and granule release. Moreover, P2Y6 stimulated the function of mast cells through activation of the AKT signaling pathway. Our data provides evidence that P2Y6 contributes to allergic airway inflammation and remodeling by enhancing the functions of mast cells in ovalbumin-induced asthmatic mice.

Topics: Animals; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Chemotaxis; Cytokines; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Mast Cells; Mice; Mice, Inbred C57BL; Ovalbumin; Phenotype; Real-Time Polymerase Chain Reaction; Receptors, Purinergic P2; Signal Transduction; Uridine Diphosphate

Eosinophils are native to the healthy gastrointestinal tract and are associated with inflammatory diseases likely triggered by exposure to food allergens (e.g., food allergies and eosinophilic gastrointestinal disorders). In models of allergic respiratory diseases and in vitro studies, direct Ag engagement elicits eosinophil effector functions, including degranulation and Ag presentation. However, it was not known whether intestinal tissue eosinophils that are separated from luminal food Ags by a columnar epithelium might similarly engage food Ags. Using an intestinal ligated loop model in mice, in this study we determined that resident intestinal eosinophils acquire Ag from the lumen of Ag-sensitized but not naive mice in vivo. Ag acquisition was Ig-dependent; intestinal eosinophils were unable to acquire Ag in sensitized Ig-deficient mice, and passive immunization with immune serum or Ag-specific IgG was sufficient to enable intestinal eosinophils in otherwise naive mice to acquire Ag in vivo. Intestinal eosinophils expressed low-affinity IgG receptors, and the activating receptor FcγRIII was necessary for Ig-mediated acquisition of Ags by isolated intestinal eosinophils in vitro. Our combined data suggest that intestinal eosinophils acquire lumen-derived food Ags in sensitized mice via FcγRIII Ag focusing and that they may therefore participate in Ag-driven secondary immune responses to oral Ags.

Topics: Adaptive Immunity; Allergens; Animals; Antigen Presentation; Antigens; Cells, Cultured; Eosinophils; Hypersensitivity; Immunity, Humoral; Immunoglobulin E; Intestine, Small; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Models, Animal; Ovalbumin; Receptors, IgG

Skin is protected by a tough but flexible multilayered barrier and is a front line for immune responses against invading particles. For many years now, skin has been a tissue where certain vaccines are injected for the prevention of infectious disease, however, the detailed mechanisms of the skin immune response are not yet well understood. Using thin and small injection needles, we carefully injected OVA into a restricted region of mouse skin, i.e., intradermal (ID), and examined the antibody response in comparison with subcutaneous (SC) injection or epicutaneous patch administration of OVA. Epicutaneous patches induced a high IgE response against OVA, but IgG production was low. High IgG production was induced by both ID and SC injection, moreover, ID injection induced higher IgG production without any adjutants. Furthermore, OVA-specific IgE production was diminished by ID injection. We found that ID injection could efficiently stimulate skin resident DCs, drive Th1-biased conditions and diminish IgE production. The ID injection response was regulated by Langerin+ dermal DCs, because OVA was taken up mainly by these cells and, after transiently deleting them, the IgE response was no longer diminished and IgG1 production was enhanced. We also tested whether ID injection might be an effective allergy treatment by attempting to inhibit ongoing IgE production in mice with experimentally induced high serum IgE levels. Multiple ID injections of OVA were shown to prevent elevation of serum OVA-specific IgE after repeated allergen challenge. In contrast, SC OVA injection could only transiently inhibit the OVA-specific IgE production. These findings indicated that ID injection results in higher induction of antigen-specific IgG, and thus may be useful for vaccine delivery with little or no adjuvant components. Moreover, the observed diminishment of IgE and induction of Th1-biased immune responses suggest that ID may be a useful injection route for allergy immunotherapy.

Topics: Allergens; Animals; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunohistochemistry; Immunotherapy; Injections, Intradermal; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Skin; Th1 Cells; Ultrasonography; Vaccines

Histamine is a mast cell mediator released e.g. during food allergy. The aim of the project was to identify the effect of histamine on rat submucosal neurons and the mechanisms involved. Cultured submucosal neurons from rat colon express H1, H2 and H3 receptors as shown by immunocytochemical staining confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) with messenger RNA (mRNA) isolated from submucosal homogenates as starting material. Histamine evoked a biphasic rise of the cytosolic Ca(2+) concentration in cultured submucosal neurons, consisting in a release of intracellularly stored Ca(2+) followed by an influx from the extracellular space. Although agonists of all three receptor subtypes evoked an increase in the cytosolic Ca(2+) concentration, experiments with antagonists revealed that mainly H1 (and to a lesser degree H2) receptors mediate the response to histamine. In coculture experiments with RBL-2H3 cells, a mast cell equivalent, compound 48/80, evoked an increase in the cytosolic Ca(2+) concentration of neighbouring neurons. Like the response to native histamine, the neuronal response to the mast cell degranulator was strongly inhibited by the H1 receptor antagonist pyrilamine and reduced by the H2 receptor antagonist cimetidine. In rats sensitized against ovalbumin, exposure to the antigen induced a rise in short-circuit current (I sc) across colonic mucosa-submucosa preparations without a significant increase in paracellular fluorescein fluxes. Pyrilamine strongly inhibited the increase in I sc, a weaker inhibition was observed after blockade of protease receptors or 5-lipoxygenase. Consequently, H1 receptors on submucosal neurons seem to play a pivotal role in the communication between mast cells and the enteric nervous system.

Topics: Animals; Calcium Signaling; Cells, Cultured; Coculture Techniques; Colon; Disease Models, Animal; Enteric Nervous System; Female; Histamine; Histamine Agonists; Histamine Antagonists; Hypersensitivity; Intestinal Mucosa; Male; Mast Cells; Membrane Potentials; Neurons; Ovalbumin; Paracrine Communication; Rats, Wistar; Receptors, Histamine; RNA, Messenger

Eosinophils accumulate at the site of allergic inflammation and are critical effector cells in allergic diseases. Recent studies have also suggested a role for eosinophils in the resolution of inflammation.. To determine the role of eosinophils in the resolution phase of the response to repeated allergen challenge.. Eosinophil-deficient (PHIL) and wild-type (WT) littermates were sensitized and challenged to ovalbumin (OVA) 7 or 11 times. Airway inflammation, airway hyperresponsiveness (AHR) to inhaled methacholine, bronchoalveolar lavage (BAL) cytokine levels, and lung histology were monitored. Intracellular cytokine levels in BAL leukocytes were analyzed by flow cytometry. Groups of OVA-sensitized PHIL mice received bone marrow from WT or IL-10(-/-) donors 30 days before the OVA challenge.. PHIL and WT mice developed similar levels of AHR and numbers of leukocytes and cytokine levels in BAL fluid after OVA sensitization and 7 airway challenges; no eosinophils were detected in the PHIL mice. Unlike WT mice, sensitized PHIL mice maintained AHR, lung inflammation, and increased levels of IL-4, IL-5, and IL-13 in BAL fluid after 11 challenges whereas IL-10 and TGF-β levels were decreased. Restoration of eosinophil numbers after injection of bone marrow from WT but not IL-10-deficient mice restored levels of IL-10 and TGF-β in BAL fluid as well as suppressed AHR and inflammation. Intracellular staining of BAL leukocytes revealed the capacity of eosinophils to produce IL-10.. After repeated allergen challenge, eosinophils appeared not essential for the development of AHR and lung inflammation but contributed to the resolution of AHR and inflammation by producing IL-10.

Topics: Allergens; Animals; Bone Marrow Transplantation; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Leukocyte Count; Lung; Male; Mice; Mice, Transgenic; Ovalbumin; Respiratory Hypersensitivity

In the last 20 years, the development of murine models of allergic asthma has provided researchers with a means to explore the mechanisms of this T-helper type 2 (Th2)-driven inflammatory disease. While systemic sensitization and airway challenge with ovalbumin has been the most widely used model, recent emphasis has been placed on the development of models using more naturally occurring antigens. However, the diversity of models currently available makes it hard for investigators new to this field to choose to use the most effective and appropriate model to test their hypothesis. Here we describe three different mouse models of allergic asthma, including the classical ovalbumin model, a modified ovalbumin model that has been shown to be mast-cell dependent, as well as a house dust mite antigen-induced model. We also discuss briefly their characterization and differences, in the aim to facilitate the choice of the appropriate model when working on this intricate Th2 inflammatory disease.

Topics: Animals; Asthma; Bronchoalveolar Lavage; Disease Models, Animal; Flow Cytometry; Hypersensitivity; Leukocyte Count; Lung; Mice; Ovalbumin; Pyroglyphidae

Ginger has been used commonly in the traditional system of medicine for the treatment of respiratory disorders.. The present study investigates the immunosuppressive activity of ginger by using the mouse model of ovalbumin-induced allergic asthma.. Treatment with ethanol extract (500 mg/kg) and aqueous extract (720 mg/kg) of rhizomes, and methylprednisolone (5 mg/kg) was initiated 1 week after second sensitization of mice with ovalbumin and continued for 7 d. RT-PCR followed by gel electrophoresis and ELISA were used for the evaluation of mRNA expression levels and protein levels of Th2 type markers, respectively. Lung tissue histopathology was conducted by using H&E and PAS staining.. We observed significant reduction in goblet cell hyperplasia (0.83 ± 0.17 and 1.0 ± 0.26), infiltration of inflammatory cells in airways (0.67 ± 0.33 and 1.0 ± 0.37), and edema with vascular congestion (1.0 ± 0.26 and 1.2 ± 0.17) by both ethanol and aqueous extracts, respectively. A highly significant reduction of total and differential count of eosinophils and neutrophils in BALF, and eosinophil count in blood were also observed. Both extracts significantly inhibited Th2-mediated immune response, which is evident by a decrease in mRNA expression levels of IL-4 and IL-5. Protein levels of IL-4 and IL-5 in BALF, along with total serum IgE levels, were also significantly suppressed by both extracts.. Our study validated the traditional use of ginger in respiratory disorders and suggests that ginger reduces allergic airway inflammation, possibly by the suppression of Th2-mediated immune response.

Topics: Animals; Asthma; Hypersensitivity; Immunity, Cellular; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Th2 Cells; Zingiber officinale

Clinical studies reveal platelet activation in patients with asthma, allergic rhinitis, and eczema. This is distinct from platelet aggregation, which is critical for the maintenance of hemostasis and in which a role for platelet purinergic receptors is well documented. However, purines are also essential for inflammatory cell trafficking in animal models of allergic lung inflammation, which are known to be platelet dependent, yet the role of purines in the platelet activation accompanying inflammation is unknown.. We investigated whether the involvement of purine activation of platelets during allergic inflammation is distinct from purine involvement in platelet aggregation.. BALB/c mice were sensitized to ovalbumin and subsequent airway ovalbumin challenge. Bronchoalveolar lavage fluid was analyzed for inflammatory cells, and blood samples were assessed for platelet activation. The role of platelet purinergic receptors and associated signaling mechanisms (RhoA) were assessed.. P2Y₁, but not P2Y₁₂ or P2X₁, antagonism inhibited pulmonary leukocyte recruitment. The formation of platelet-leukocyte complexes in vivo and platelet/P-selectin-dependent polymorphonuclear cell migration in vitro were exclusively platelet P2Y₁ receptor dependent. Furthermore, platelet P2Y₁ activation resulted in RhoA activity in vivo after allergen challenge, and RhoA signaling in platelets through P2Y₁ stimulation was required for platelet-dependent leukocyte chemotaxis in vitro. Leukocyte recruitment in thrombocytopenic mice remained suppressed after reinfusion of platelets pretreated with a P2Y₁ antagonist or a Rho-associated kinase 1 inhibitor, confirming the crucial role of platelet P2Y₁ receptor and subsequent activation of RhoA.. RhoA signaling downstream of platelet P2Y₁, but not P2Y₁₂, represents a clear dichotomy in platelet activation during allergic inflammation versus hemostasis.

Topics: Adenosine Diphosphate; Allergens; Animals; Blood Platelets; Chemokines; Chemotaxis, Leukocyte; Disease Models, Animal; Female; Hypersensitivity; Leukocytes; Lung; Macrophages; Mice; Models, Biological; Ovalbumin; P-Selectin; Platelet Aggregation; Purinergic P2Y Receptor Agonists; Receptors, Purinergic P2Y1; rhoA GTP-Binding Protein; Signal Transduction

In our previous studies, the recombinant type II macrophage migration inhibitory factor homologue (rAs-MIF) secreted from Anisakis simplex suppressed experimental inflammation mouse model through IL-10 production and CD4(+)CD25(+)Foxp3(+) T-cell recruitment. Also, TLR2 gene expression was significantly increased following rAs-MIF treatment. To know the relation between TLR2 and amelioration mechanisms of rAs-MIF, we induced allergic airway inflammation by ovalbumin and alum with or without rAs-MIF under TLR2 blocking systems [anti-TLR2-specific antibody (α-mTLR2 Ab) treatment and using TLR2 knockout mice]. As a result, the amelioration effects of rAs-MIF in allergic airway inflammation model (diminished inflammation and Th2 response in the lung, increased IL-10 secretion, CD4(+)CD25(+)Foxp3(+) T-cell recruitment) were diminished under two of the TLR2 blocking model. The expression of TLR2 on the surface of lung epithelial cell was significantly elevated by rAs-MIF treatment or Pam3CSK (TLR2-specific agonist) treatment, but they might have some competition effect on the elevation of TLR2 expression. In addition, the elevation of IL-10 gene expression by rAs-MIF treatment was significantly inhibited by α-mTLR2 Ab or Pam3CSK pretreatment. In conclusion, anti-inflammatory effects of the rAs-MIF on OVA-induced allergic airway inflammation might be closely related to TLR2.

Topics: Alum Compounds; Animals; Anisakis; Disease Models, Animal; Female; Helminth Proteins; Hypersensitivity; Inflammation; Interleukin-10; Lung; Macrophage Migration-Inhibitory Factors; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory; Toll-Like Receptor 2

Dendritic cells play an important role in determining whether naïve T cells mature into either Th1 or Th2 cells. We determined whether heat-shock protein X (HspX) purified from Mycobacterium tuberculosis regulates the Th1/Th2 immune response in an ovalbumin (OVA)-induced murine model of asthma. HspX increased interferon-gamma, IL-17A, -12 and transforming growth factor (TGF)-β production and T-bet gene expression but reduced IL-13 production and GATA-3 gene expression. HspX also inhibited asthmatic reactions as demonstrated by an increase in the number of eosinophils in bronchoalveolar lavage fluid, inflammatory cell infiltration in lung tissues, airway luminal narrowing, and airway hyper-responsiveness. Furthermore, HspX enhanced OVA-induced decrease of regulatory T cells in the mediastinal lymph nodes. This study provides evidence that HspX plays critical roles in the amelioration of asthmatic inflammation in mice. These findings provide new insights into the immunotherapeutic role of HspX with respect to its effects on a murine model of asthma.

Topics: Adoptive Transfer; Animals; Asthma; Dendritic Cells; GATA3 Transcription Factor; Heat-Shock Proteins; Hypersensitivity; Immunotherapy; Inflammation Mediators; Lung; Mice; Mycobacterium tuberculosis; Ovalbumin

Colonization with Streptococcus pneumoniae (S. pneumoniae) is associated with an increased risk for recurrent wheeze and asthma. Killed S. pneumoniae showed some potential as an effective immunomodulatory therapy in a murine model of asthma. Murine studies demonstrated protection against allergic asthma by symbiotic bacteria via triggering regulatory T cell response: treatment with killed S. pneumoniae resulted in suppressed levels of allergen-specific Th2 cytokines, while early immunization generated a protective Th1 response. We investigated the impact of lung infection with live S. pneumoniae on both the development and maintenance of allergic airway inflammation and respiratory tolerance in mice. BALB/c mice were infected intratracheally with S. pneumoniae either prior to or after tolerance or allergy were induced, using ovalbumin (OVA) as model allergen. Infection of mice with S. pneumoniae prior to sensitization or after manifestation of allergic airway inflammation suppressed the development of an allergic phenotype as judged by reduced eosinophil counts in bronchoalveolar lavage fluid, decreased IgE serum levels and Th2 cytokines, relative to non-infected allergic control mice. In contrast, infection of mice with S. pneumoniae after manifestation of allergic airway inflammation combined with late mucosal re-challenge did not affect the allergic response. Moreover, induction and maintenance of respiratory tolerance to OVA challenge were not altered in S. pneumoniae-infected mice, demonstrating that mice remained tolerant to the model allergen and were protected from the development of allergic airway inflammation regardless of the time point of infection. Our results suggest that a bacterial infection may decrease the manifestation of an allergic phenotype not only prior to sensitization but also after manifestation of allergic airway inflammation in mice, whereas both, induction and maintenance of respiratory tolerance are not affected by pneumococcal pneumonia. These data may point to a role for undisturbed development and maintenance of mucosal tolerance for the prevention of allergic inflammation also in humans.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin E; Immunoglobulin G; Mice; Ovalbumin; Phenotype; Pneumonia, Pneumococcal; Respiratory Mucosa; Respiratory System; Streptococcus pneumoniae

Eosinophils are key effector cells in allergic diseases, including allergic rhinitis, eczema, and asthma. Their tissue presence is regulated by both recruitment and increased longevity at inflamed sites.. To investigate the ability of the flavone wogonin to induce eosinophil apoptosis in vitro and attenuate eosinophil-dominant allergic inflammation in vivo in mice.. Human and mouse eosinophil apoptosis in response to wogonin was investigated by cellular morphology, flow cytometry, mitochondrial membrane permeability, and pharmacological caspase inhibition. Allergic lung inflammation was modeled in mice sensitized and challenged with ovalbumin. Bronchoalveolar lavage (BAL) and lung tissue were examined for inflammation, mucus production, and inflammatory mediator production. Airway hyperresponsiveness to aerosolized methacholine was measured.. Wogonin induced time- and concentration-dependent human and mouse eosinophil apoptosis in vitro. Wogonin-induced eosinophil apoptosis occurred with activation of caspase-3 and was inhibited by pharmacological caspase inhibition. Wogonin administration attenuated allergic airway inflammation in vivo with reductions in BAL and interstitial eosinophil numbers, increased eosinophil apoptosis, reduced airway mucus production, and attenuated airway hyperresponsiveness. This wogonin-induced reduction in allergic airway inflammation was prevented by concurrent caspase inhibition in vivo.. Wogonin induces eosinophil apoptosis and attenuates allergic airway inflammation, suggesting that it has therapeutic potential for the treatment of allergic inflammation in humans.

Topics: Animals; Apoptosis; Bronchoalveolar Lavage; Eosinophils; Female; Flavanones; Flow Cytometry; Humans; Hypersensitivity; In Vitro Techniques; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin

Chronic inflammatory disorder of the airways causes asthma. Regulatory T cells (Treg cells) and Natural killer T cells (NKT cells) both play critical roles in the pathogenesis of asthma. Activation of Treg cells requires Foxp3, whereas whether Foxp3 may regulate the ratio of Treg and NKT cells to affect asthma is uncertain. In an ovalbumin (OVA)-induced mouse model of asthma, we either increased Treg cells by lentivirus-mediated forced expression of exogenous Foxp3, or increased NKT cells by stimulation with its activator α-GalCer. We found that the CD4+CD25+ Treg cells increased by forced Foxp3 expression, and decreased by α-GalCer, while the CD3+CD161+ NKT cells decreased by forced Foxp3 expression, and increased by α-GalCer. Moreover, forced Foxp3 expression, but not α-GalCer, significantly alleviated the hallmarks of asthma. Furthermore, forced Foxp3 increased levels of IL_10 and TGFβ1, and α-GalCer increased levels of IL_4 and INFγ in the OVA-treated lung. Taken together, our study suggests that Foxp3 may activate Treg cells and suppress NKT cells in asthma. Treg and NKT cells may antagonize the effects of each other in asthma.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Forkhead Transcription Factors; Galactosylceramides; HEK293 Cells; Humans; Hypersensitivity; Lentivirus; Male; Mice, Inbred C57BL; Natural Killer T-Cells; Ovalbumin; T-Lymphocytes, Regulatory

Lunasin is a naturally occurring peptide isolated from soybeans and has been explored in cancer treatment. Lunasin inhibits NF-κB activation and thus pro-inflammatory cytokine and mediator production in macrophages. In this study we demonstrate that lunasin can effectively suppress allergic airway inflammation in two murine models of asthma. In an OVA+Alum sensitization model, intranasal lunasin treatment at the time of OVA challenges significantly reduced total cells counts in bronchoalveolar lavage (BAL) fluid and eosinophilia, peribronchiolar inflammatory infiltration, goblet cell metaplasia and airway IL-4 production. In an OVA+LPS intranasal sensitization model, lunasin treatment either at the time of sensitization or challenge has similar effects in suppress allergic airway inflammation including significantly reduced total cell and eosinophil counts in BAL fluid, inflammatory gene Fizz1 expression in the lung, and IL-4 production by OVA re-stimulated cells from mediastinal lymph nodes. We further show that intranasal instillation of OVA+lunasin significantly increases OVA-specific regulatory T cell (Treg) accumulation in the lung comparing to OVA only treatment. Taken together, our results suggest lunasin as an anti-inflammatory agent can be potentially used in asthma therapy or as an adjuvant to enhance the induction of antigen-specific Tregs and thus boost the efficacy of allergy immunotherapy.

Topics: Alum Compounds; Animals; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Goblet Cells; Hypersensitivity; Lipopolysaccharides; Lung; Mice; Ovalbumin; Soybean Proteins; T-Lymphocytes, Regulatory

Synthesis and SAR of a series of 7-azaindoles as Orai channel inhibitors showing good potency inhibiting IL-2 production in Jurkat cells is described. Compound 14d displaying best pharmacokinetic properties was further characterized in a model of allergen induced asthma showing inhibition in the number of eosinophils in BALF. High lipophilicity remains as one of the main challenges for this class of compounds.

Topics: Animals; Asthma; Aza Compounds; Calcium Channel Blockers; Calcium Channels; Disease Models, Animal; Drug Evaluation, Preclinical; Half-Life; Humans; Hypersensitivity; Indoles; Interleukin-2; Jurkat Cells; Microsomes; Models, Biological; Ovalbumin; Protein Binding; Pyridines; Pyrroles; Rats; Structure-Activity Relationship

The murine intestinal nematode Heligmosomoides polygyrus exerts multiple immunomodulatory effects in the host, including the suppression of allergic inflammation in mice sensitized to allergen presented with alum adjuvant. Similar suppression is attained by co-administration of H. polygyrus excretory/secretory products (HES) with the sensitizing dose of ovalbumin (OVA) in alum. We investigated the mechanism of suppression by HES in this model, and found it was maintained in MyD88xTRIF-deficient mice, implying no role for helminth- or host-derived TLR ligands, or IL-1 family cytokines that signal in a MyD88- or TRIF-dependent manner. We also found suppression was unchanged in µMT mice, which lack B2 B cells, and that suppression was not abrogated when regulatory T cells were depleted in Foxp3.LuciDTR-4 mice. However, reduced IL-5 production was seen in the first 12 h after injection of OVA-alum when HES was co-administered, associated with reduced activation of IL-5(+) and IL-13(+) group 2 innate lymphoid cells. Thus, the suppressive effects of HES on alum-mediated OVA sensitization are reflected in the very earliest innate response to allergen exposure in vivo.

Topics: Adaptor Proteins, Vesicular Transport; Adjuvants, Immunologic; Alum Compounds; Animals; B-Lymphocytes, Regulatory; Hypersensitivity; Immunologic Deficiency Syndromes; Lymphocytes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Nematospiroides dubius; Ovalbumin; Primary Immunodeficiency Diseases; Signal Transduction; T-Lymphocytes, Regulatory

B cells have been described as having the capacity to regulate cellular immune responses and suppress inflammatory processes. One such regulatory B-cell population is defined as IL-10-producing CD19(+) CD1d(hi) cells. Previous work has identified an expansion of these cells in mice infected with the helminth, Schistosoma mansoni. Here, microarray analysis of CD19(+) CD1d(hi) B cells from mice infected with S. mansoni demonstrated significantly increased Tlr7 expression, while CD19(+) CD1d(hi) B cells from uninfected mice also demonstrated elevated Tlr7 expression. Using IL-10 reporter, Il10(-/-) and Tlr7(-/-) mice, we formally demonstrate that TLR7 ligation of CD19(+) CD1d(hi) B cells increases their capacity to produce IL-10. In a mouse model of allergic lung inflammation, the adoptive transfer of TLR7-elicited CD19(+) CD1d(hi) B cells reduced airway inflammation and associated airway hyperresponsiveness. Using DEREG mice to deplete FoxP3(+) T regulatory cells in allergen-sensitized mice, we show that that TLR7-elicited CD19(+) CD1d(hi) B cells suppress airway hyperresponsiveness via a T regulatory cell dependent mechanism. These studies identify that TLR7 stimulation leads to the expansion of IL-10-producing CD19(+) CD1d(hi) B cells, which can suppress allergic lung inflammation via T regulatory cells.

Topics: Animals; Antigens, CD19; Antigens, CD1d; B-Lymphocytes; Disease Models, Animal; Humans; Hypersensitivity; Interleukin-10; Mice; Mice, Knockout; Ovalbumin; Pneumonia; Protein Binding; Respiratory Hypersensitivity; Schistosoma mansoni; Schistosomiasis mansoni; T-Lymphocytes, Regulatory; Toll-Like Receptor 7; Up-Regulation

Comorbid asthma in sickle cell disease (SCD) confers higher rates of vaso-occlusive pain and mortality, yet the physiological link between these two distinct diseases remains puzzling. We used a mouse model of SCD to study pulmonary immunology and physiology before and after the induction of allergic airway disease (AAD). SCD mice were sensitized with ovalbumin (OVA) and aluminum hydroxide by the intraperitoneal route followed by daily, nose-only OVA-aerosol challenge to induce AAD. The lungs of naive SCD mice showed signs of inflammatory and immune processes: (1) histologic and cytochemical evidence of airway inflammation compared with naive wild-type mice; (2) bronchoalveolar lavage (BAL) fluid contained increased total lymphocytes, %CD8+ T cells, granulocyte-colony stimulating factor, interleukin 5 (IL-5), IL-7, and chemokine (C-X-C motif) ligand (CXCL)1; and (3) lung tissue and hilar lymph node (HLN) had increased CD4+, CD8+, and regulatory T (Treg) cells. Furthermore, SCD mice at AAD demonstrated significant changes compared with the naive state: (1) BAL fluid with increased %CD4+ T cells and Treg cells, lower %CD8+ T cells, and decreased interferon gamma, CXCL10, chemokine (C-C motif) ligand 2, and IL-17; (2) serum with increased OVA-specific immunoglobulin E, IL-6, and IL-13, and decreased IL-1α and CXCL10; (3) no increase in Treg cells in the lung tissue or HLN; and (4) hyporesponsiveness to methacholine challenge. In conclusion, SCD mice have an altered immunologic pulmonary milieu and physiological responsiveness. These findings suggest that the clinical phenotype of AAD in SCD mice differs from that of wild-type mice and that individuals with SCD may also have a unique, divergent phenotype perhaps amenable to a different therapeutic approach.

Topics: Anemia, Sickle Cell; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Female; Hemizygote; Hypersensitivity; Inflammation; Leukocyte Count; Lung; Mice, Inbred C57BL; Mucus; Ovalbumin; T-Lymphocytes

Recent evidence has suggested that miRNA is implicated in the immune response of allergic and inflammatory diseases. However, little is known about its role in the mechanism that underlies the establishment of pro-allergic DCs in allergic rhinitis is not fully understood. This study assessed whether and how microRNA (miR)-106b regulates the pro-allergic properties of DCs upon allergen stimulation in vitro. Bone marrow-derived dendritic cells (BMDCs) were generated and stimulated with ovalbumin (OVA) to identify the miRNA expression profile. After transfection with miR-106b mimics and inhibitors OVA-activated BMDCs were further evaluated for surface marker expression using flow cytometry, cytokine production using ELISA and subsequent effects on Th2 cell polarisation using flow cytometry. Moreover, the upstream controllers and potential target proteins of miR-106b were examined in a western blot analysis. Results showed that MiR-106b expression was significantly inhibited in activated BMDCs upon OVA stimulation (p<0.05). Surface marker expression (e.g., MHC class II, CD80 and CD86) was significantly upregulated after the transfection of an miR-106b inhibitor (p<0.05), and the proportion of GATA-3(+) T cells was significantly increased among CD4(+) T cells that were cocultured with miR-106b inhibitor-pretreated BMDCs (p<0.05). Conversely, IL-12 production from OVA-activated BMDCs and the proportion of T-bet(+) T cells increased significantly in a coculture of CD4(+) T cells and miR-106b mimics-transfected BMDCs (p<0.05). The early growth response (Egr)-2 was identified via luciferase reporter assays as a target gene of miR-106b, and significant Egr-2 upregulation was observed in OVA-activated BMDCs following transfection with a miR-106b inhibitor (p<0.05). In conclusion, our results suggest that miR-106b negatively regulates the pro-allergic properties of BMDCs and subsequent Th2 polarisation upon OVA stimulation and might represent a promising therapeutic target for allergic inflammation.

Topics: Allergens; Animals; Cells, Cultured; Dendritic Cells; Early Growth Response Protein 2; Hypersensitivity; Male; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Th2 Cells

To evaluate the antiallergic effect of newly characterised probiotic strains, Lactobacillus fermentum NWS29, Lactobacillus casei NWP08 and Lactobacillus rhamnosus NWP13, mice were divided into six experimental groups: control, ovalbumin (OVA), NWS29, NWP08, NWP13 and L. rhamnosus GG (LGG). Mice were immunised and probiotics were administered via oral gavage followed by challenge with OVA. After last challenge with OVA, inflammatory cells in bronchoalveolar lavage fluid (BALF), recruitment of inflammatory cells in airways and OVA-specific immunoglobulin E (IgE) in serum were determined by Giemsa, haematoxylin and eosin (HE) staining, and ELISA, respectively. Relative mRNA expression of interleukins (IL-4, IL-5, IL-10, IL-13 and IL-17), transforming growth factor-β (TGF-β) and interferon-γ (IFN-γ) in lung and spleen tissue was determined by real time RT-PCR. OVA-specific IgE levels, recruitment of eosinophils and mRNA expressions of inflammatory cytokines were remarkably increased in OVA-exposed mice compared with the control group. Administration of NWS29 and NWP13 suppressed inflammatory cell infiltration in airways and BALF, and level of OVA-specific IgE in serum of OVA-exposed mice. Furthermore, NWS29 and NWP13 also abrogated the mRNA expression of 1L-4, IL-5, IL-13 and TGF-β in mice immunised and exposed to OVA. Our findings suggest that NWS29 and NWP13 might be good candidates for the prevention of allergic airway inflammation.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cytokines; Cytological Techniques; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Gene Expression Profiling; Histocytochemistry; Hypersensitivity; Immunoglobulin E; Immunologic Factors; Lacticaseibacillus casei; Lacticaseibacillus rhamnosus; Limosilactobacillus fermentum; Lung; Mice, Inbred BALB C; Ovalbumin; Probiotics; Spleen; Treatment Outcome

Allergic asthma is a chronic inflammatory disorder that affects ∼20% of the population worldwide. Microarray analyses of nasal epithelial cells from acute asthmatic patients detected a 50% decrease in expression of Stard7, an intracellular phosphatidylcholine transport protein. To determine whether loss of Stard7 expression promotes allergic responses, mice were generated in which one allele of the Stard7 locus was globally disrupted (Stard7 (+/-) mice). OVA sensitization and challenge of Stard7(+/-) mice resulted in a significant increase in pulmonary inflammation, mucous cell metaplasia, airway hyperresponsiveness, and OVA-specific IgE compared with OVA-sensitized/challenged wild-type (WT) mice. This exacerbation was largely Th2-mediated with a significant increase in CD4(+)IL-13(+) T cells and IL-4, IL-5, and IL-13 cytokines. The loss of Stard7 was also associated with increased lung epithelial permeability and activation of proinflammatory dendritic cells in sensitized and/or challenged Stard7 (+/-) mice. Notably, OVA-pulsed dendritic cells from Stard7(+/-) mice were sufficient to confer an exaggerated allergic response in OVA-challenged WT mice, although airway hyperresponsiveness was greater in Stard7(+/-) recipients compared with WT recipients. Enhanced allergic responses in the lung were accompanied by age-dependent development of spontaneous atopic dermatitis. Overall, these data suggest that Stard7 is an important component of a novel protective pathway in tissues exposed to the extracellular environment.

Topics: Adoptive Transfer; Animals; Carrier Proteins; Cytokines; Dendritic Cells; Dermatitis; Disease Models, Animal; Disease Progression; Female; Gene Deletion; Haploinsufficiency; Hypersensitivity; Lung; Male; Mice; Mice, Knockout; Models, Biological; Ovalbumin; Permeability; Respiratory Mucosa; Skin; Th2 Cells

Eosinophils are hallmark cells of allergic Th2 respiratory inflammation. However, the relative importance of eosinophil activation and the induction of effector functions such as the expression of IL-13 to allergic Th2 pulmonary disease remain to be defined.. Wild-type or cytokine-deficient (IL-13(-/-) or IL-4(-/-) ) eosinophils treated with cytokines (GM-CSF, IL-4, IL-33) were adoptively transferred into eosinophil-deficient recipient mice subjected to allergen provocation using established models of respiratory inflammation. Allergen-induced pulmonary changes were assessed.. In contrast to the transfer of untreated blood eosinophils to the lungs of recipient eosinophil deficient mice, which induced no immune/inflammatory changes either in the lung or in the lung draining lymph nodes (LDLN), pretreatment of blood eosinophils with GM-CSF prior to transfer elicited trafficking of these eosinophils to LDLN. In turn, these LDLN eosinophils elicited the accumulation of dendritic cells and CD4(+) T cells to these same LDLNs without inducing pulmonary inflammation. However, exposure of eosinophils to GM-CSF, IL-4, and IL-33 prior to transfer induced not only immune events in the LDLN, but also allergen-mediated increases in airway Th2 cytokine/chemokine levels, the subsequent accumulation of CD4(+) T cells as well as alternatively activated (M2) macrophages, and the induction of pulmonary histopathologies. Significantly, this allergic respiratory inflammation was dependent on eosinophil-derived IL-13, whereas IL-4 expression by eosinophils had no significant role.. The data demonstrate the differential activation of eosinophils as a function of cytokine exposure and suggest that eosinophil-specific IL-13 expression by activated cells is a necessary component of the subsequent allergic Th2 pulmonary pathologies.

Topics: Allergens; Animals; Cells, Cultured; Cytokines; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Interleukin-13; Lung; Macrophages; Male; Mice; Mice, Transgenic; Ovalbumin; Phenotype; Th2 Cells

Bu-zhong-yi-qi-tang (BZYQT), an herbal formula of traditional Chinese medicine, has been an effective regimen of allergic diseases for nearly 800 years. Our previous report has demonstrated its anti-inflammatory effects in patients with perennial allergic rhinitis, and the aim of this study is to investigate the anti-asthmatic effect of BZYQT.. Female BALB/cByJNarl mice were sensitized with normal saline (control group) or OVA. Mice sensitized by OVA were fed with distilled water (OVA group), oral 0.5 g/Kg (low-dose group) or 1 g/Kg (high-dose group) of BZYQT solution once daily on days 36-40 besides their routine diet. Airway hyper-responsiveness (AHR), eosinophil infiltration, levels of cytokines and total immunoglobulin E (IgE) in broncho-alveolar lavage fluid (BALF) were determined. The lungs and tracheas were removed, and histopathologic examination was subsequently performed.. AHR was significantly reduced in both low- and high-dose BZYQT groups compared with the OVA group after inhalation of the highest dose of methacholine (50 mg/ml). The levels of eotaxin, Th2-related cytokines (IL-4, IL-5, IL-13), IgE, and eosinophil infiltration in BALF were significantly decreased in both BZYQT groups compared with the OVA group. Histopathologic examination revealed that eosinophil infiltration of the lung and trachea tissues was remarkably attenuated in both BZYQT groups.. Oral administration of BZYQT solution may exert anti-asthmatic effect by relieving AHR in OVA-sensitized mice, which is compatible with our clinical experience. Although detailed mechanism is to be determined, we surmise that it may be correlated with the immune-modulatory effects of inhibiting Th2 responses on the basis of our limited results.

Topics: Administration, Oral; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Immunologic Factors; Lung; Mice, Inbred BALB C; Ovalbumin

Heparanase is an endo-β-glucuronidase that specifically cleaves heparan sulfate proteoglycans in the extracellular matrix. Expression of this enzyme is increased in several pathological conditions including inflammation. We have investigated the role of heparanase in pulmonary inflammation in the context of allergic and non-allergic pulmonary cell recruitment using heparanase knockout (Hpa-/-) mice as a model. Following local delivery of LPS or zymosan, no significant difference was found in the recruitment of neutrophils to the lung between Hpa-/- and wild type (WT) control. Similarly neutrophil recruitment was not inhibited in WT mice treated with a heparanase inhibitor. However, in allergic inflammatory models, Hpa-/- mice displayed a significantly reduced eosinophil (but not neutrophil) recruitment to the airways and this was also associated with a reduction in allergen-induced bronchial hyperresponsiveness, indicating that heparanase expression is associated with allergic reactions. This was further demonstrated by pharmacological treatment with a heparanase inhibitor in the WT allergic mice. Examination of lung specimens from patients with different severity of chronic obstructive pulmonary disease (COPD) found increased heparanase expression. Thus, it is established that heparanase contributes to allergen-induced eosinophil recruitment to the lung and could provide a novel therapeutic target for the development of anti-inflammatory drugs for the treatment of asthma and other allergic diseases.

Topics: Animals; Enzyme Inhibitors; Female; Gene Deletion; Glucuronidase; Humans; Hypersensitivity; Immunization; Inflammation; Lung; Male; Mice, Inbred BALB C; Middle Aged; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Respiratory Function Tests

The mammalian target of rapamycin (mTOR) signalling pathway regulates immune responses, and promotes cell growth and differentiation. Inhibition of mTOR with rapamycin modulates allergic asthma, while the underlying molecular mechanisms remain elusive. Here, we demonstrate that rapamycin, effectively inhibits eosinophil differentiation, contributing to its overall protective role in allergic airway inflammation.. Rapamycin was administered in a mouse model of ovalbumin-induced allergic airway inflammation, and the eosinophil differentiation was analysed in vivo and in vitro.. Rapamycin significantly attenuated allergic airway inflammation and markedly decreased the amount of eosinophils in local airways, peripheral blood and bone marrow, independently of levels of interleukin-5 (IL-5). In vitro colony forming unit assay and liquid culture demonstrated that rapamycin directly inhibited IL-5-induced eosinophil differentiation. In addition, rapamycin reduced the production of IL-6 and IL-13 by eosinophils. Rapamycin was also capable of reducing the eosinophil levels in IL-5 transgenic NJ.1638 mice, again regardless of the constitutive high levels of IL-5. Interestingly, rapamycin inhibition of eosinophil differentiation in turn resulted in an accumulation of eosinophil lineage-committed progenitors in bone marrow.. Altogether these results clearly demonstrate a direct inhibitory role of rapamycin in eosinophil differentiation and function, and reemphasize the importance of rapamycin and possibly, mTOR, in allergic airway disease.

Topics: Animals; Asthma; Cell Differentiation; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunosuppressive Agents; Inflammation; Interleukins; Leukocyte Count; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Serine Proteinase Inhibitors; Signal Transduction; Sirolimus; TOR Serine-Threonine Kinases

Feasibility of microneedles (MNs) for cutaneous allergen specific immunotherapy (ASI) is demonstrated by comparing against currently practiced subcutaneous (SC) allergen immunotherapy, and the intramuscular (IM) and intraperitoneal (IP) routes. In Balb/c mice with ovalbumin (Ova, 25 μg) as the allergen MNs-Ova without alum induced anti-Ova IgG response comparable to IM but higher than SC and IP groups (250 μg alum was additionally used for SC, IM and IP groups). MNs-Ova induced higher anti-Ova IgG1 and IgG2a responses in comparison to other routes; however IgG2b and IgG3 responses were significantly lower than the IP group. As in SC group, anti-Ova IgE and IgA were low for MNs-Ova. Furthermore, MNs-Ova induced expression of IL-5, IL-13, IFN-γ and IL-1β cytokines in serum, but at significantly lower levels than other routes. Overall, MNs-Ova induced allergen-specific IgG antibodies, and activated the Th1 pathway (evidenced by higher IgG2a levels), suggesting their potential use for painless ASI.

Topics: Adjuvants, Immunologic; Administration, Cutaneous; Allergens; Animals; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Injections, Intramuscular; Injections, Intraperitoneal; Mice, Inbred BALB C; Ovalbumin; Th1 Cells

c-Jun N-terminal kinase (JNK) relays extracellular stimuli through phosphorylation cascades that lead to various cell responses. In the present study, we aimed to investigate the effect of the JNK inhibitor SP600125 on the resolution of airway inflammation, and the underlying mechanism using a murine acute asthma model.. Female C57BL/6 mice were sensitized with saline or ovalbumin (OVA) on day 0, and challenged with OVA on day 14-20. Meanwhile, some of the mice were treated with SP600125 (30 mg/kg) intraperitoneally 2 h before each challenge. The airway inflammation was evaluated by counting the numbers of various types of inflammatory cells in bronchoalveolar lavage fluid (BALF), histopathology, cytokines production and mucus secretion in individual mouse. In addition, we analyzed the protein levels of phosphorylated JNK and TLR9 in the lung tissues.. SP600125 markedly reduced the invasion of inflammatory cells into the peribronchial regions, and decreased the numbers of eosinophils, monocytes, neutrophils and lymphocytes in BALF. SP600125 also reduced the level of plasma OVA-specific IgE, lowered the production of pro-inflammatory cytokines in BALF and alleviated mucus secretion. Meanwhile, SP600125 inhibited OVA-induced, increased expression of p-JNK and TLR9 in the lung tissues.. Collectively, our data demonstrated that SP600125 promoted resolution of allergic airway inflammation via TLR9 in an OVA-induced murine acute asthma model. The JNK-TLR9 pathway may be a new therapeutic target in the treatment for the allergic asthma.

Topics: Acute Disease; Animals; Anthracenes; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; JNK Mitogen-Activated Protein Kinases; Lung; Mice, Inbred C57BL; Mucus; Ovalbumin; Phosphorylation; Toll-Like Receptor 9

 Macrophages include the classically activated pro-inflammatory M1 macrophages (M1s) and alternatively activated anti-inflammatory M2 macrophages (M2s). The M1s are activated by both interferon-γ and Toll-like receptor ligands, including lipopolysaccharide (LPS), and have potent pro-inflammatory activity. In contrast, Th2 cytokines activate the M2s, which are involved in the immune response to parasites, promotion of tissue remodeling, and immune regulatory functions. Although alveolar macrophages (AMs) play an essential role in the pulmonary immune system, little is known about their phenotypes..  Quantitative reverse transcription polymerase chain reaction and flow cytometry were used to define the characteristics of alveolar macrophages derived from untreated naïve mice and from murine models of both ovalbumin (OVA)-induced allergic airway inflammation and LPS-induced acute airway inflammation. AMs were co-cultured with CD4(+) T cells and were pulsed with tritiated thymidine to assess proliferative responses..  We characterized in detail murine AMs and found that these cells were not completely consistent with the current M1 versus M2-polarization model. OVA-induced allergic and LPS-induced acute airway inflammation promoted the polarization of AMs towards the current M2-skewed and M1-skewed phenotypes, respectively. Moreover, our data also show that CD11c(+) CD11b(+) AMs from the LPS-treated mice play a regulatory role in antigen-specific T-cell proliferation in vitro..  These characteristics of AMs depend on the incoming pathogens they encounter and on the phase of inflammation and do not correspond to the current M1 versus M2-polarization model. These findings may facilitate an understanding of their contributions to the pulmonary immune system in airway inflammation.

Topics: Allergens; Animals; Asthma; CD4-Positive T-Lymphocytes; Cell Proliferation; Cytokines; Female; Hypersensitivity; Inflammation; Lipopolysaccharides; Lung; Macrophages, Alveolar; Mice; Mice, Inbred BALB C; Ovalbumin

Orally administered probiotic micro-organisms are able to regulate the exacerbated immune response during the antigenic sensitisation process. The aim of the present study was to evaluate the potential efficacy of probiotic fermented milk (PFM) in preventing or treating allergy in an experimental model, and to investigate its underlying mechanisms. Ovoalbumin (OVA)-sensitised BALB/c mice were fed with PFM before the sensitisation procedure or fed continuously with PFM. At 7 and 15 d post-sensitisation, anti-OVA-specific IgE, IgG, IgG1 and IgG2a concentrations were measured in the serum and broncho-alveolar lavage fluid (BALF). Concentrations of interferon-γ (IFN-γ), IL-4, IL-10 and total secretory IgA (S-IgA) were measured in the supernatants of macerated lungs or in the BALF. The levels of IgA+, CD4+ and CD8+ T lymphocytes and F4/80+ cells were measured in the lungs by immunofluorescence. Inducible CD4+/CD25/Foxp3+ regulatory T (Treg) cells were evaluated in the lungs. PFM shifted the T helper (Th)2 profile response towards a Th1 response that led to the production of IgG instead of IgE, with increasing levels of IL-10 and IFN-γ that play an important role in immunomodulation exerted by PFM administration in sensitised mice. Anti-OVA-specific IgE levels were significantly decreased; however, there was no modification in the levels of anti-OVA-specific IgG and total S-IgA. PFM did not influence Treg cells in treated mice. Consumption of PFM could be a promising strategy in the amelioration of airway allergies, considering that the effect is mediated by the production of IgG through the activation of Th1 instead of the direct activation of Th2 cells to produce IgE.

Topics: Animals; Bacteria; Bronchoalveolar Lavage Fluid; Cultured Milk Products; Cytokines; Diet; Fermentation; Hypersensitivity; Immunoglobulin G; Immunoglobulins; Lung; Male; Mice, Inbred BALB C; Ovalbumin; Probiotics; T-Lymphocytes, Regulatory; Th1 Cells; Th1-Th2 Balance; Th2 Cells

Lipopolysaccharide (LPS) is ubiquitous in the environment and can therefore, exacerbate allergic responses. Studies have suggested immunoregulatory effects of LPS according to route, dose and stage of exposure. Present study has examined whether dose and stage of LPS exposure (during sensitization and challenge with OVA) exacerbates airway inflammations, antigen specific-IgE level, histamine release, Th1/Th2 cytokine response. Further, anti-asthmatic potential of curcumin, through intranasal route has been evaluated for the first time in LPS induced airway inflammation in an ovalbumin (OVA)-challenged mouse asthma model.. Balb/c mice were first sensitized with OVA on 1st and 8th day and exposed to two LPS doses (0.1/1.0 μg) separately on 2nd day and then further exposed to LPS with OVA-aerosol (from 9 to 14 day). Further, lower LPS dose (0.1 μg) was chosen for OVA exposed mouse model of asthma exacerbation study. Intranasal curcumin was administered from 9th to 14th day before every LPS exposure.. Exposure to LPS (0.1 μg) exacerbates airway inflammations in terms of IgE level, Th2-cytokine response (IL-4 and IL-5), histamine release, EPO and MPO activities and oxidative stress. Intranasal curcumin has effectively ameliorated airway exacerbations whereas dexamethasone, a known glucocorticosteroid, was not promising as compared to intranasal curcumin.. Schedule and dose of LPS exposure determines asthma exacerbations and intranasal curcumin could be better immunomodulatory agent in LPS exposed asthma exacerbations.

Topics: Administration, Intranasal; Animals; Asthma; Curcumin; Cytokines; Histamine; Hypersensitivity; Immunoglobulin E; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress

Airway mucus hypersecretion is a key pathophysiological feature in asthma. Fasudil, a selective Rho-A/Rho kinase inhibitor, has been used in clinical trials to treat pulmonary hypertension. However, its function in modulating airway mucus hypersecretion in asthma remains undefined.. We examined whether fasudil, a selective Rho-A/Rho kinase inhibitor, affects the mucus hypersecretion by suppressing MUC5AC via signal transducer and activator of transcription factor 6 (STAT6) and nuclear factor-kappa B (NFκB) in mice and cells.. We measured mucus secretion and the expression of Rho-kinase in the airway tissue of patients with asthma. BALB/c mice were sensitized and challenged with ovalbumin (OVA) followed with fasudil treatment. The lung tissues were assessed for airway inflammation and mucus secretion. Cytokine levels and airway responsiveness were determined. STAT6 and NFκB were quantified by Western blot. 16HBE cells were stimulated with house dust mite (HDM) extracts. MUC5AC and muc5ac promoter activities were measured. Using siRNA to knockdown STAT6 in epithelial cells, we determined the impact of STAT6 on muc5ac promoter activity. NFκB nuclear translocation was observed with immunostaining.. Fasudil administration significantly decreased the number of inflammatory cells, inflammation index in the lung and airway responsiveness. Fasudil also reduced mucous secretion and MUC5AC expression in OVA-challenged mice. Fasudil down-regulated the levels of IL-17, IL-4 and IL-13 in the lung tissue of OVA-challenged mice. Fasudil also decreased the expression and phosphorylation of NFκB and STAT6 as well as the nuclear translocation of NFκB. In addition, human airway epithelial cells (16HBE) were challenged with HDM extracts and then treated with fasudil. Fasudil inhibited HDM extract-induced MUC5AC expression, which is associated with a reduction in STAT6 and NFκB in epithelial cells.. These findings indicate that the Rho-A/Rho kinase inhibitor, fasudil, plays a negative regulatory role in allergen-induced mucus secretion and MUC5AC expression by regulating STAT6 and NFκB.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Allergens; Animals; Asthma; Case-Control Studies; Cell Line; Cytokines; Disease Models, Animal; Down-Regulation; Female; Gene Expression; Humans; Hypersensitivity; Male; Mice; Mucin 5AC; Mucus; NF-kappa B; Ovalbumin; Protein Kinase Inhibitors; Pyroglyphidae; Respiratory Mucosa; rho-Associated Kinases; Signal Transduction; STAT6 Transcription Factor

Aluminum salts gels (alum) are TLR-independent adjuvants and have been used to boost antibody responses in alum-based vaccines such as diphtheria, pertussis, and tetanus toxoid (DPT) triple vaccine. However, the pro-Th2 activity of alum-based vaccine formulations has not been fully appreciated. Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies. The adsorption into alum of prototypic TLR4 agonists such as lipopolysaccharides (LPS) derived from Escherichia coli consistently dampened TT-induced Th2 activities without inducing IFNγ or Th1-like responses in the lung. Conversely, adsorption of monophosphoryl lipid A (MPLA) extracted from Salmonella minnesota, which is a TIR-domain-containing adapter-inducing interferon-β- (TRIF-) biased TLR4 agonist, was less effective in decreasing Th-2 responses. Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization. Notably, LPS increased the production of IgG1 and IgG2a TT-specific antibodies. In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

Topics: Adjuvants, Immunologic; Adsorption; Alum Compounds; Animals; Antibody Formation; Antigens, Bacterial; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Hypersensitivity; Immunization; Immunoglobulin E; Lipopolysaccharides; Mice; Ovalbumin; Poly I-C; Tetanus Antitoxin; Tetanus Toxoid; Th2 Cells; Toll-Like Receptor 3; Toll-Like Receptor 4

Food allergy can influence the development of colorectal cancer, although the underlying mechanisms are unclear. While mast cells (MC) store and secrete histamine, immature myeloid cells (IMC) are the major site of histidine decarboxylase (HDC) expression, the enzyme responsible for histamine production. From our earlier work, we hypothesized that histamine is central to the association between allergy and colorectal carcinogenesis through its influence on the MC-MDSC axis. Here, we show that in wild type (WT) mice, ovalbumin (OVA) immunization elicits a typical TH2 response. In contrast, in HDC-/- mice, the response to OVA allergy is skewed towards infiltration by IL-17 expressing MCs. This response is inhibited by histamine treatment. The HDC-/- allergic IL-17-expressing MCs promote MDSC proliferation and upregulation of Cox-2 and Arg-1. OVA allergy in HDC-/- mice increases the growth of colon tumor cells in both the MC38 tumor cell implantation model and the AOM/DSS carcinogenesis model. Taken together, our results show that histamine represses IL-17-expressing MCs and their subsequent activation of MDSCs, attenuating the risk of colorectal cancer in the setting of food allergy. Targeting the MC-MDSC axis may be useful for cancer prevention and treatment in patients, particularly in those with food allergy.

Topics: Amino Acid Sequence; Animals; Colorectal Neoplasms; Disease Models, Animal; Female; Histamine; Hypersensitivity; Interleukin-17; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Sequence Data; Myeloid Cells; Ovalbumin; Peptide Fragments

Oxyresveratrol is known to possess anti-inflammatory property. Current study investigates the immunosuppressive effect of oxyresveratrol by using mouse model of ovalbumin (OVA)-induced allergic airway inflammation.. BALB/c mice were randomly divided into five groups having 8 mice in each group. Treatment with low dose (7 mg/kg) and high dose (15 mg/kg) of oxyresveratrol, and methylprednisolone (15 mg/kg; standard drug) was started 2 week after immunization of mice with ovalbumin and continued for 7d. Ovalbumin was also injected into pinna of right ear 24h before sacrificing the mice to evaluate delayed type hypersensitivity (DTH). H&E and PAS staining were used for histopathological evaluation of lungs. Reverse transcription polymerase chain reaction followed by gel electrophoresis were used for evaluation of mRNA expression levels of IL-4, IL-5, and IL-13.. Oxyresveratrol significantly reduced total leucocyte count in both blood and bronchoalveolar lavage fluid (BALF). Treatment with oxyresveratrol normalized altered eosinophil and neutrophil counts in both blood and BALF. OVA-specific T-cell response was also significantly inhibited by oxyresveratrol. A significant attenuation of inflammatory cell infiltration and goblet cell hyperplasia was observed after treatment with oxyresveratrol. Data showed that oxyresveratrol significantly suppressed Th2 (T helper cells) type immune response which was obvious by the reduction in mRNA expression levels of IL-4, IL-5, and IL-13. Similarly, treatment with methylprednisolone also significantly reduced all the immunomodulatory and inflammatory parameters.. Our study demonstrates that oxyresveratrol ameliorates allergic asthma. The anti-asthmatic activity might in part occur via the down regulation of IL-4, IL-5, and IL-13 expression levels.

Topics: Animals; Bronchoalveolar Lavage Fluid; Hypersensitivity; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Stilbenes; T-Lymphocytes

Surfactant protein D (SP-D) is a pulmonary collectin important in lung immunity. SP-D-deficient mice (Sftpd(-/-)) are reported to be susceptible to ovalbumin (OVA)- and fungal allergen-induced pulmonary inflammation, while treatment with exogenous SP-D has therapeutic effects in such disease models. β-Glucans are a diverse group of polysaccharides previously suggested to serve as fungal ligands for SP-D. We set out to investigate if SP-D could interact with 1,3-β-glucan and attenuate allergic pulmonary inflammation in the presence of 1,3-β-glucan. Allergic airway disease was induced in Sftpd(-/-) and Sftpd(+/+) mice by OVA sensitization and subsequent challenge with OVA, 1,3-β-glucan, or OVA/1,3-β-glucan together. Mice in the combined treatment group were further treated with a high dose of recombinant fragment of human SP-D (rfhSP-D). We demonstrated direct interaction between SP-D and 1,3-β-glucan. OVA-induced mucous cell metaplasia was increased in Sftpd(-/-) mice, supporting previously reported protective effects of endogenous SP-D in allergy. OVA-induced parenchymal CCL11 levels and eosinophilic infiltration in bronchoalveolar lavage were unaffected by 1,3-β-glucan, but were reversed with rfhSP-D treatment. 1,3-β-Glucan treatment did, however, induce pulmonary neutrophilic infiltration and increased TNF-α levels in bronchoalveolar lavage, independently of OVA-induced allergy. This infiltration was also reversed by treatment with rfhSP-D. 1,3-β-Glucan reduced OVA-induced mucous cell metaplasia, T helper 2 cytokines, and IFN-γ production. rfhSP-D treatment further reduced mucous metaplasia and T helper 2 cytokine secretion to background levels. In summary, rfhSP-D treatment resulted in attenuation of both allergic inflammation and 1,3-β-glucan-mediated neutrophilic inflammation. Our data suggest that treatment with high-dose SP-D protects from mold-induced exacerbations of allergic asthma.

Topics: Animals; beta-Glucans; Chemokine CCL11; Cytokines; Female; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Ligands; Metaplasia; Mice, Inbred C57BL; Microbiota; Ovalbumin; Protective Agents; Proteoglycans; Pulmonary Alveoli; Pulmonary Surfactant-Associated Protein D; Respiratory Hypersensitivity

Mast cells and basophils are effector cells in the pathophysiology of allergic diseases. Targeted elimination of these cells may be a promising strategy for the treatment of allergic disorders. Our present study aims at targeted delivery of anti-FcεRIα Fab-conjugated celastrol-loaded micelles toward FcεRIα receptors expressed on mast cells and basophils to have enhanced anti-allergic effect. To achieve this aim, we prepared celastrol-loaded (PEO-block-PPO-block-PEO, Pluronic) polymeric nanomicelles using thin-film hydration method. The anti-FcεRIα Fab Fragment was then conjugated to carboxyl groups on drug-loaded micelles via EDC amidation reaction. The anti-FcεRIα Fab-conjugated celastrol-loaded micelles revealed uniform particle size (93.43 ± 12.93 nm) with high loading percentage (21.2 ± 1.5% w/w). The image of micelles showed oval and rod like. The anti-FcεRIα Fab-conjugated micelles demonstrated enhanced cellular uptake and cytotoxity toward target KU812 cells than non-conjugated micelles in vitro. Furthermore, diffusion of the drug into the cells allowed an efficient induction of cell apoptosis. In mouse model of allergic asthma, treatment with anti-FcεRIα Fab-conjugated micelles increased lung accumulation of micelles, and significantly reduced OVA-sIgE, histamine and Th2 cytokines (IL-4, IL-5, TNF-α) levels, eosinophils infiltration and mucus production. In addition, in mouse model of passive cutaneous anaphylaxis, anti-FcεRIα Fab-conjugated celastrol-loaded micelles treatment significantly decreased extravasated evan's in the ear. These results indicate that anti-FcεRIα Fab-conjugated celastrol-loaded micelles can target and selectively kill mast cells and basophils which express FcεRIα, and may be efficient reagents for the treatment of allergic disorders and mast cell related diseases.

Topics: Animals; Apoptosis; Asthma; Basophils; Biological Transport; Cell Line; Drug Carriers; Drug Liberation; Hypersensitivity; Immunoglobulin Fab Fragments; Mast Cells; Mice; Micelles; Ovalbumin; Particle Size; Passive Cutaneous Anaphylaxis; Pentacyclic Triterpenes; Receptors, IgE; Tissue Distribution; Triterpenes

The protective effect of infectious agents against allergic reactions has been thoroughly investigated. Current studies have demonstrated the ability of some helminths to modulate the immune response of infected hosts. The objective of the present study was to investigate the relationship between Toxocara canis infection and the development of an allergic response in mice immunised with ovalbumin (OVA). We determined the total and differential blood and bronchoalveolar lavage fluid cells using BALB/c mice as a model. To this end, the levels of interleukin (IL)-4, IL-5 and IL-10 and anti-OVA-IgE were measured using an ELISA. The inflammatory process in the lungs was observed using histology slides stained with haematoxylin and eosin. The results showed an increase in the total number of leukocytes and eosinophils in the blood of infected and immunised animals at 18 days after infection. We observed a slight lymphocytic inflammatory infiltrate in the portal space in all infected mice. Anti-OVA-IgE levels were detected in smaller proportions in the plasma of immunised and infected mice compared with mice that were only infected. Therefore, we concluded that T. canis potentiates inflammation in the lungs in response to OVA, although anti-OVA-IgE levels suggest a potential reduction of the inflammatory process through this mechanism.

Topics: Animals; Antibodies; Biopsy; Bronchoalveolar Lavage Fluid; Enzyme-Linked Immunosorbent Assay; Eosinophils; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-10; Interleukin-4; Interleukin-5; Leukocyte Count; Lung; Mice, Inbred BALB C; Ovalbumin; Toxocara canis; Toxocariasis

Activated macrophages have been classified into classical (M1) and alternative (M2) macrophages. We aimed to establish a method to yield enough number of macrophages to analyze their molecular, biological and immunological functions. We used drugs; adjuvant albumin from chicken egg whites--Imject Alum (OVA-Alum) and OVA Complete Freund Adjuvant (OVA-CFA), to induce macrophages to M2 and M1 respectively. We analyzed the phenotype of purified macrophages induced under these immune conditions, using flow cytometry (FACS) to detect cell-surface molecules and the enzyme-linked immunosorbent assay (ELISA) was used to detect cytokines. The cDNA microarray was employed to measure changes in expression level of cell surface protein between M1 and M2 macrophages. Phenotype analysis of purified macrophages, induced under these immune conditions, showed macrophages induced by OVA-Alum was almost M2 while the proportion of M1 macrophages induced by OVA-CFA was significantly higher. The results also showed higher expression level of macrophage galactose N- acetyl-galactosamine specific lectin-2 protein (MGL1/2-PE), a known M2 macrophage marker, on the surface of Alum-induced macrophages. On the basis of these preliminary data, ELISA results revealed that after macrophage stimulation with lipopolysaccharides (LPS), the level of interleukin (IL)-10 produced by Alum- induced macrophages was higher than the level of IL-10 produced by CFA-induced macrophages. In contrast, the level of tumor necrosis factor-alpha (TNF-α) produced by CFA-induced macrophages was higher than Alum-induced macrophages. The cDNA microarray confirmed previous results and suggest immunoglobulin-like type 2 receptor alpha (Pilra) as a new marker for M1, macrophage galactose N-acetylgalactosamine-specific lectin 2 (Mgl2) as M2 macrophages marker.

Topics: Alum Compounds; Animals; Cell Polarity; Freund's Adjuvant; Hypersensitivity; Macrophages; Male; Mice; Oligonucleotide Array Sequence Analysis; Ovalbumin; Th1 Cells; Th2 Cells

Sulfate-modifying factor 2 (SUMF2), a member of the formylglycine-generating enzyme family, was earlier found to play a role in the regulation of interleukin (IL)-13 expression and secretion in airway smooth muscle cells. IL-13 is a T helper 2 cytokine that plays important roles in the pathogenesis of asthma. However, there is little evidence of the potential role of SUMF2 in the cellular inflammatory responses in asthma. Here, using an ovalbumin-induced asthma rat model, we show that SUMF2 gene expression is significantly decreased in allergic asthma rats. Moreover, several pathological changes were observed in the lung tissue and IL-13 was found to be overexpressed in the ovalbumin-induced asthma model. Additional studies on the lung bronchial epithelial tissues, peripheral blood lymphocytes and bronchoalveolar lavage fluid of the OVA-induced asthma rats showed that SUMF2 mRNA and protein expression were attenuated. However, there was only a little significant correlation was found between SUMF2 and IL-13 expression. These results indicate that SUMF2 may mediate airway inflammation in allergic asthma by modulating the expression of IL-13. More data from in vivo experiments are needed to clearly understand the role of SUMF2 in asthma.

Topics: Allergens; Animals; Asthma; Blotting, Western; Disease Models, Animal; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression Regulation; Hypersensitivity; Inflammation; Interleukin-13; Ovalbumin; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction; Sulfatases; Transcriptome

Matrine has been isolated from Sophora flavescens, and found to show anti-inflammatory effects in macrophages and anti-cachectic effects in hepatomas. The present study investigated whether matrine suppressed eosinophil infiltration and airway hyperresponsiveness (AHR) in mice, and decreased the inflammatory response of tracheal epithelial cells.. BALB/c mice were sensitized and challenged with ovalbumin to induce allergic asthma in mice. These asthmatic mice were given various doses of matrine by intraperitoneal injection. Additionally, activated human tracheal epithelial cells (BEAS-2B cells) were treated with matrine, and evaluated for levels of proinflammatory cytokines and chemokines.. We found that matrine significantly decreased AHR, and suppressed goblet cell hyperplasia, eosinophil infiltration, and inflammatory response in the lung tissue of asthmatic mice. Matrine also reduced the levels of Th2 cytokines and chemokines in bronchoalveolar lavage fluid, and suppressed OVA-IgE production in serum. Furthermore, matrine treatment of activated BEAS-2B cells decreased production of proinflammatory cytokines and eotaxins, as well as suppressed ICAM-1 expression and thus adhesion of eosinophils to inflammatory BEAS-2B cells in vitro.. Our findings suggest that matrine can improve allergic asthma in mice, and therefore has potential therapeutic potential in humans.

Topics: Alkaloids; Animals; Asthma; Cell Adhesion; Cell Line; Cytokines; Eosinophils; Gene Expression Regulation; Goblet Cells; Humans; Hypersensitivity; Lung; Matrines; Mice; Mice, Inbred BALB C; Ovalbumin; Quinolizines

Perillyl alcohol (POH) is an isoprenoid which inhibits farnesyl transferase and geranylgeranyl transferase, key enzymes that induce conformational and functional changes in small G proteins to conduct signal production for cell proliferation. Thus, it has been tried for the treatment of cancers. However, although it affects the proliferation of immunocytes, its influence on immune responses has been examined in only a few studies. Notably, its effect on antigen-induced immune responses has not been studied. In this study, we examined whether POH suppresses Ag-induced immune responses with a mouse model of allergic airway inflammation. POH treatment of sensitized mice suppressed proliferation and cytokine production in Ag-stimulated spleen cells or CD4(+) T cells. Further, sensitized mice received aerosolized OVA to induce allergic airway inflammation, and some mice received POH treatment. POH significantly suppressed indicators of allergic airway inflammation such as airway eosinophilia. Cytokine production in thoracic lymph nodes was also significantly suppressed. These results demonstrate that POH suppresses antigen-induced immune responses in the lung. Considering that it exists naturally, POH could be a novel preventive or therapeutic option for immunologic lung disorders such as asthma with minimal side effects.

Topics: Animals; Antigens; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Hypersensitivity; Immunosuppression Therapy; Immunosuppressive Agents; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Monoterpenes; Ovalbumin; Pulmonary Eosinophilia

Endogenous mediators, such as RvE1 (resolvin E1), promote resolution of an inflammatory response and have potential as novel therapeutic agents. In the present study, we investigated the activity of RvE1 in a model of an acute exacerbation of chronic allergic asthma in mice. Animals sensitized to OVA (ovalbumin) received controlled low-level challenge with aerosolized antigen for 4 weeks, followed by a single moderate-level challenge to simulate an allergen-induced exacerbation of asthmatic inflammation. Induction of an exacerbation was associated with rapid recruitment of neutrophils, lymphocytes and eosinophils, together with increased levels of Th2 and pro-inflammatory cytokines. When administered before the final moderate-level challenge, RvE1 had only a modest effect on airway inflammation. To assess its effects when administered after induction of an exacerbation, we first characterized the cellular and molecular events associated with spontaneous resolution of airway inflammation over the following 96 h. Subsequently, we showed that administration of RvE1 at 2 and 8 h after the final challenge accelerated this process significantly. Specifically, RvE1 promoted a decline in the number of inflammatory cells, concentration of cytokines in lavage fluid and expression of mRNA for cytokines by macrophages, confirming its pro-resolution activity. In vitro, RvE1 had no apparent effect on lymphocytes, but suppressed significantly cytokine production by pulmonary macrophages, with evidence of down-regulation of the nuclear translocation of NF-κB (nuclear factor κB) p65 in these cells. The present study provides novel evidence that RvE1 can facilitate resolution of airway inflammation in a clinically relevant model of an acute exacerbation of asthma, possibly via its effects on activated pulmonary macrophages.

Topics: Active Transport, Cell Nucleus; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Disease Models, Animal; Eicosapentaenoic Acid; Female; Hypersensitivity; Immunohistochemistry; Inflammation; Leukocytes; Macrophages; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; RNA, Messenger; Time Factors

Agglomerated carbon black nanoparticles (CBNPs) administered via respiratory or subcutaneous routes have been shown to promote allergic sensitization to coadministered ovalbumin (OVA) protein in rodents. In the present study, we aimed to model and elucidate the mechanism of this adjuvanticity using an in vitro assay based on T cell sensitization to ovalbumin₃₂₃₋₃₃₉ peptide (OVA(p)). CBNP base particles of 22 and 39 nm were characterized and termed CBNP22 and CBNP39 powders. Splenic leukocytes derived from transgenic DO11.10 mice were exposed to suspensions of media alone, concanavalin A mitogen, CBNP agglomerates smaller than 220 nm, OVA(p) alone, OVA(p) + anti-CD28 costimulant, OVA(p) + cyclosporin A immunosuppressant, or OVA(p) + CBNPs. Samples were analyzed at 72 h post-exposure. Proliferation rate, a marker of cellular mitosis, was assessed. Polymerase chain reaction arrays were used to assess genes involved in allergic response pathways. The mitogen control, costimulatory control, and immunosuppressive control chemicals modified the T helper cell proliferation rate. CBNP22 mildly reduced proliferation at 12 μg/ml, but CBNP39 did not. Gene expression analysis of cells treated with OVA(p) showed that coincubation with 12 μg/ml CBNP22 enhanced gene expression of interleukin-4 (IL-4), IL-10, and IL-13, all allergy-associated Th2 cytokines. Coincubation of OVA(p) with 12 μg/ml CBNP39 significantly enhanced IL-13 gene expression concurrent with downregulation of the Th1-associated transcription factor Stat4. IL-4 and IL-13 protein secretion reflected the mRNA trends. The changes were consistently higher in cells exposed to CBNP22 than CBNP39, suggesting that smaller particle size, higher surface area, and higher purity were associated with the direct adjuvant effect on Th2 cells in this genetically susceptible model of OVA allergy.

Topics: Adjuvants, Immunologic; Animals; Cell Proliferation; Cells, Cultured; Cytokines; Female; Flow Cytometry; Hypersensitivity; Male; Mice; Mice, Inbred Strains; Microscopy, Electron, Scanning; Nanoparticles; Ovalbumin; Particle Size; Porosity; Primary Cell Culture; Soot; Surface Properties; Th2 Cells

Low-level laser therapy (LLLT) controls bronchial hyperresponsiveness (BHR) associated with increased RhoA expression as well as pro-inflammatory mediators associated with NF-kB in acute lung inflammation. Herein, we explore if LLLT can reduce both BHR and Th2 cytokines in allergic asthma. Mice were studied for bronchial reactivity and lung inflammation after antigen challenge. BHR was measured through dose-response curves to acetylcholine. Some animals were pretreated with a RhoA inhibitor before the antigen. LLLT (660 nm, 30 mW and 5.4 J) was applied on the skin over the right upper bronchus and two irradiation protocols were used. Reduction of BHR post LLLT coincided with lower RhoA expression in bronchial muscle as well as reduction in eosinophils and eotaxin. LLLT also diminished ICAM expression and Th2 cytokines as well as signal transducer and activator of transduction 6 (STAT6) levels in lungs from challenged mice. Our results demonstrated that LLLT reduced BHR via RhoA and lessened allergic lung inflammation via STAT6.

Topics: Airway Remodeling; Amides; Animals; Asthma; Bronchi; Bronchial Hyperreactivity; Bronchoconstriction; Cytokines; Enzyme Inhibitors; Hypersensitivity; Low-Level Light Therapy; Lung; Male; Mice; Mice, Inbred BALB C; Muscle, Smooth; Ovalbumin; Pneumonia; Pyridines; rho GTP-Binding Proteins; rhoA GTP-Binding Protein; STAT6 Transcription Factor

Peptide immunotherapy (PIT) offers realistic prospects for the treatment of allergic diseases, including allergic asthma. Much is understood of the behavior of naive T cells in response to PIT. However, treatment of patients with ongoing allergic disease requires detailed understanding of the responses of allergen-experienced T cells. CD62L expression by allergen-experienced T cells corresponds to effector/effector memory (CD62L(lo)) and central memory (CD62L(hi)) subsets, which vary with allergen exposure (e.g., during, or out with, pollen season). The efficacy of PIT on different T helper 2 (Th2) cell memory populations is unknown. We developed a murine model of PIT in allergic airway inflammation (AAI) driven by adoptively transferred, traceable ovalbumin-experienced Th2 cells. PIT effectively suppressed AAI driven by unfractionated Th2 cells. Selective transfer of CD62L(hi) and CD62L(lo) Th2 cells revealed that these two populations behaved differently from one another and from previously characterized (early deletional) responses of naive CD4(+) T cells to PIT. Most notably, allergen-reactive CD62L(lo) Th2 cells were long-lived within the lung after PIT, before allergen challenge, in contrast to CD62L(hi) Th2 cells. Despite this, PIT was most potent against CD62L(lo) Th2 cells in protecting from AAI, impairing their ability to produce Th2 cytokines, whereas this capacity was heightened in PIT-treated CD62L(hi) Th2 cells. We conclude that Th2 cells do not undergo an early deletional form of tolerance after PIT. Moreover, memory Th2 subsets respond differently to PIT. These findings have implications for the clinical translation of PIT in different allergic scenarios.

Topics: Animals; Bronchoalveolar Lavage; Flow Cytometry; Hypersensitivity; Immunologic Memory; Immunotherapy; L-Selectin; Lung; Mice; Mice, Transgenic; Ovalbumin; Peptide Fragments; Th2 Cells

Skullcap (Scutellaria baicalensis) has been widely used as a dietary ingredient and traditional herbal medicine owing to its anti-inflammatory and anticancer properties. In this study, we investigated the anti-allergic effects of skullcap and its active compounds, focusing on T cell-mediated responses ex vivo and in vivo. Splenocytes from mice sensitized with ovalbumin (OVA) were isolated for analyses of cytokine production and cell viability. Mice sensitized with OVA were orally administered skullcap or wogonin for 16 days, and then immunoglobulin (Ig) and cytokine levels were measured by enzyme-linked immunosorbent assays. Treatment with skullcap significantly inhibited interleukin (IL)-4 production without reduction of cell viability. Moreover, wogonin, but not baicalin and baicalein, suppressed IL-4 and interferon-gamma production. In vivo, skullcap and wogonin downregulated OVA-induced Th2 immune responses, especially IgE and IL-5 prediction. Wogonin as an active component of skullcap may be applied as a therapeutic agent for IgE- and IL-5-mediated allergic disorders.

Topics: Animals; Anti-Allergic Agents; Flavanones; Hypersensitivity; Immunity, Innate; Mice; Ovalbumin; Phytotherapy; Plant Extracts; Scutellaria baicalensis; Th2 Cells

Maternal immunization with allergens, such as ovalbumin (OVA), can inhibit the development of an allergic response in offspring. The regulatory mechanisms seem to be mediated by maternal antibodies (MatAbs) and factors generated by the maternal-fetal interface. The aim of this study was to verify the pathways of inhibitory Ab transference after maternal immunization with OVA and the effect of the offspring's dendritic cells (DCs) on the generation of regulatory T (Treg) cells. We verified that preconceptional OVA immunization induces high levels of proinflammatory and regulatory cytokines in the amniotic fluid, allowing the transference of high levels of anti-OVA IgG1 Abs to the offspring. Using an adoptive nursing protocol, we verified that maternal immunization leads to MatAb transference by the placental route and by breastfeeding contribute to the inhibition of anaphylactic IgE and IgG1 Ab responses in immunized offspring. We observed that maternal immunization decreased eosinophil numbers in recovered bronchoalveolar lavage fluid and in the lung tissue, whereas with a lack of control of airway responsiveness to methacholine. Maternal immunization induced in young offspring a decreased percentage of CD11c+ DCs expressing MHC class II and CD40 molecules. Moreover, DCs from both groups of offspring when pulsed with OVA, were able to induce Treg cells in vitro. Similarly, OVA immunization at the neonatal stage increased the frequency of Treg cells, regardless of the mother's immunization status. These findings emphasize that maternal immunization leads to a complex interaction of regulatory factors, with MatAbs, DCs and Treg cells affecting the tolerance of offspring during an allergic response.

Topics: Allergens; Amniotic Fluid; Animals; Animals, Newborn; Bronchoalveolar Lavage Fluid; Cytokines; Dendritic Cells; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin E; Immunoglobulin G; Maternal Exposure; Mice; Ovalbumin; Rats; T-Lymphocyte Subsets

Epidemiological evidence suggesting that exposure to traffic air pollution may enhance sensitization to common allergens in children is increasing, and animal studies support biological plausibility and causality. The effect of air pollution on respiratory symptoms was suggested to be gender dependent. Previous studies showed that allergy-promoting activity of polystyrene particles (PSP) increased with decreasing particle size after footpad injection of mice. The primary aim of this study was to confirm the influence of particle size on the immunoglobulin E (IgE)-promoting capacity of particles in an airway allergy model. A second aim was to examine whether the allergy-promoting capacity of particles was influenced by gender. Female and male mice were intranasally exposed to the allergen ovalbumin (OVA) with or without ultrafine, fine, or coarse PSP modeling the core of ambient air particles. After intranasal booster immunizations with OVA, serum levels of OVA-specific IgE antibodies, and also markers of airway inflammation and cellular responses in the lung-draining mediastinal lymph nodes (MLN), were determined. PSP of all sizes promoted allergic responses, measured as increased serum concentrations of OVA-specific IgE antibodies. Further, PSP produced eosinophilic airway inflammation and elevated MLN cell numbers as well as numerically reducing the percentage of regulatory T cells. Ultrafine PSP produced stronger allergic responses to OVA than fine and coarse PSP. Although PSP enhanced sensitization in both female and male mice, significantly higher IgE levels and numbers of eosinophils were observed in females than males. However, the allergy-promoting effect of PSP was apparently independent of gender. Thus, our data support the notion that ambient air particle pollution may affect development of allergy in both female and male individuals.

Topics: Animals; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lymph Nodes; Male; Mice; Ovalbumin; Particle Size; Particulate Matter; Polystyrenes; Respiratory Hypersensitivity; Sex Factors; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Allergic asthma is characterized by chronic airway remodeling, which is associated with the expression of extracellular matrix proteins (ECM) by TGF-β. However, to date there are no reports demonstrating that structural proteins are directly expressed in mast cells. This study aimed to investigate whether ECM proteins are expressed in mast cells activated with antigen/antibody reaction, and whether the resolution effects of irradiation or 8-oxo-dG may contribute to allergic asthma prevention. Bone marrow-derived mast cells (BMMCs) were activated with DNP-HSA/anti-DNP IgE antibody (act-BMMCs). C57BL/6 mice were sensitized and challenged with ovalbumin (OVA) to induce allergic asthma. Mice were treated orally with 8-oxo-dG or exposed to whole body irradiation (using (137)Cs gamma ray at a dose of 0.5 Gy) for three consecutive days 24 h after OVA challenge. Expression of extracellular matrix (ECM) proteins, TGF-β signaling molecules and NF-κB/AP-1 was determined in the BMMCs, bronchoalveolar lavage (BAL) cells or lung tissues using Western blot, polymerase chain reaction (PCR) and electrophoretic mobility shift assay (EMSA), respectively. Act-BMMCs increased expression of ECM proteins, TGF-β/TGF-β receptor I, TGF-β signaling molecules and cytokines; and increased both NF-κB and AP-1 activity. In addition, the population of mast cells; expression of mast cell markers, TGF-β signaling molecules, ECM proteins/amounts; OVA-specific serum IgE level; numbers of goblet cells; airway hyperresponsiveness; cytokines/chemokines were increased in BAL cells and lung tissues of OVA-challenged mice. All of the above end points were reduced by irradiation or 8-oxo-dG in vitro and in vivo, respectively. The data suggest that mast cells induce expression of ECM proteins through TGF-β produced in inflammatory cells of OVA mice and that post treatment of irradiation or 8-oxo-dG after OVA-challenge may reduce airway remodeling through down-regulating mast cell re-activation by TGF-β/Smad signals.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Airway Remodeling; Animals; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Deoxyguanosine; Dose-Response Relationship, Radiation; Electrophoretic Mobility Shift Assay; Extracellular Matrix Proteins; Female; Hypersensitivity; Mast Cells; Mice; Mice, Inbred C57BL; Ovalbumin; Signal Transduction; Transforming Growth Factor beta

Ulinastatin (UTI), a serine protease inhibitor, was recently found to have an anti-inflammatory action. However, the mechanisms mediating this anti-inflammatory effect are not well understood. This study tested the hypothesis that UTI suppresses allergic inflammation by inducing the expression of haem oxygenase 1 (HO1).. Control mice and mice sensitized (on days 1, 9 and 14) and challenged (on days 21 to 27) with ovalbumin (OVA) were treated with UTI. The effects of UTI on basal expression of HO1 and that induced by OVA challenge were examined. The involvement of UTI-induced HO1 expression in anti-inflammatory and antioxidant effects of UTI was also evaluated.. UTI markedly increased basal HO1 protein expression in lungs of control mice in a time- and dose-dependent manner, and augmented HO1 protein expression induced by OVA. The up-regulation of HO1 mediated by UTI in sensitized and OVA-challenged mice was associated with reduced airway inflammation, alleviated tissue injury, reduced oxidant stress and enhanced antioxidant enzyme activities. Inhibition of HO1 activity using HO1 inhibitor, zinc protoporphyrin, attenuated inhibitory effects of UTI on inflammation and oxidant stress, and its stimulant effects on antioxidant enzyme activities. Mechanistic analysis showed that UTI increased nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), stimulated Nrf2 DNA binding activity and concomitantly up-regulated HO1 mRNA expression.. UTI is a potent and naturally occurring inducer of HO1 expression. HO1 up-regulation contributes significantly to the anti-inflammatory and organ-protective effects of UTI, which has important research and therapeutic implications.

Topics: Animals; Anti-Inflammatory Agents; Bronchoalveolar Lavage Fluid; Cytokines; Enzyme Induction; Glutathione; Glycoproteins; Heme Oxygenase-1; Hypersensitivity; Immunoglobulin E; Leukocyte Count; Lung; Malondialdehyde; Membrane Proteins; Mice; Ovalbumin; Protein Carbonylation; Superoxide Dismutase

The inflammatory immune response associated with allergic airway inflammation in asthma involves T helper type 2 (Th2) immunity. Given the data that a newly described late activator antigen-presenting cell (LAPC) population promotes Th2 immunity in viral infections, we undertook studies to investigate whether LAPCs have a pathogenic role in allergic airway inflammation.. We employed acute ovalbumin (OVA) and house dust mite (HDM) sensitization and challenge models to establish allergic airway inflammation in mice, followed by the analysis of lungs and draining lymph node (DLN) cell infiltrates, immunoglobulin E (IgE) production, and airway hyper-responsiveness (AHR). We tested whether adoptive transfer of LAPCs isolated from mice with established allergic airway inflammation augments the development of sensitization in naïve mice.. We provide evidence that in both OVA and HDM mouse models of allergic inflammation, LAPCs accumulate in the lungs and draining lymph nodes (DLNs), concomitant with the onset of lung pathology, allergen-specific IgE production, eosinophilia, and Th2 cytokine production. Adoptive transfer experiments using OVA-activated LAPCs reveal exacerbation of disease pathology with an increase in lung inflammatory cells, eosinophilia, circulating IgE, Th2 cytokine production, and a worsening of AHR. OVA-activated LAPCs preferentially increased GATA-3 induction in naïve CD4(+) T cells.. Together, these data suggest an important role for LAPCs in polarizing the Th2 response in mouse models of allergic airway inflammation.

Topics: Adoptive Transfer; Animals; Antigen-Presenting Cells; Asthma; Disease Models, Animal; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Inflammation; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Pyroglyphidae; Th2 Cells

Topics: Animals; Antigens, Plant; Hypersensitivity; Immunoglobulin E; Milk; Ovalbumin; Plant Extracts; San Francisco; United States

This study aimed to investigate the anti-allergic effects of Lactobacillus plantarum K37 (K37) on airway hyperresponsiveness (AHR) and systemic allergic responses in ovalbumin (OVA)-sensitized and -challenged BALB/c mice. Heat-inactivated K37 (105, 107, and 109 CFU/mouse, day) were orally administered to OVA-sensitized BALB/c mice to investigate their effects on AHR, immunoglobulin (Ig) and cytokine production. The results showed that K37 dose-dependently lowered the serum levels of IgE, OVA-specific IgE and OVA-specific IgG1, ameliorated AHR induced by methacholine and suppressed eosinophil infiltration in bronchoalveolar lavage fluid (BALF). The cytokine production in spleen cells culture and BALF showed that K37 drove the immune responses toward T-helper cell type 1 (Th1) responses, elevated levels of IL-2 and IFN-γ, and reduced of IL-4, IL-5 and IL-13. K37 also improved cell infiltration in lung sections. Our results demonstrated that oral administration of K37 alleviated effectively the allergic responses in vivo. Thus, K37 can be a good source material and a promising candidate for prophylactic and therapeutic treatments of allergic diseases, like asthma.

Topics: Administration, Oral; Animals; Cytokines; Female; Hypersensitivity; Immunoglobulins; Lactobacillus plantarum; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Invariant natural killer T (iNKT) cells play important immunoregulatory functions in allergen-induced airway hyperresponsiveness and inflammation. To clarify the role of iNKT cells in allergic rhinitis (AR), we generated bone marrow-derived dendritic cells (BMDCs), which were pulsed by ovalbumin (OVA) and α-galactosylceramide (OVA/α-GalCer-BMDCs) and administered into the oral submucosa of OVA-sensitized mice before nasal challenge. Nasal symptoms, level of OVA-specific immunoglobulin (IgE), and T helper type 2 (Th2) cytokine production in cervical lymph nodes (CLNs) were significantly ameliorated in wild-type (WT) mice treated with OVA/α-GalCer-BMDCs, but not in WT mice treated with OVA-BMDCs. These anti-allergic effects were not observed in Jα18(-/-) recipients that lack iNKT cells, even after similar treatment with OVA/α-GalCer-BMDCs in an adoptive transfer study with CD4(+) T cells and B cells from OVA-sensitized WT mice. In WT recipients of OVA/α-GalCer-BMDCs, the number of interleukin (IL)-21-producing iNKT cells increased significantly and the Th1/Th2 balance shifted towards the Th1 dominant state. Treatment with anti-IL-21 and anti-interferon (IFN)-γ antibodies abrogated these anti-allergic effects in mice treated with α-GalCer/OVA-BMDCs. These results suggest that activation of iNKT cells in regional lymph nodes induces anti-allergic effects through production of IL-21 or IFN-γ, and that these effects are enhanced by simultaneous stimulation with antigen. Thus, iNKT cells might be a useful target in development of new treatment strategies for AR.

Topics: Animals; B-Lymphocytes; CD4-Positive T-Lymphocytes; Dendritic Cells; Epitopes; Female; Galactosylceramides; Hypersensitivity; Immunoglobulin E; Immunotherapy, Adoptive; Interferon-gamma; Interleukins; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Natural Killer T-Cells; Ovalbumin; Rhinitis; Th1 Cells; Th2 Cells

Calcitriol (1α,25-dihydroxyvitamin D3) is the active vitamin D metabolite and mediates immunological functions, which are relevant in allergy. Its therapeutic use is limited by hypercalcaemic toxicity. We have previously shown that the activation of the vitamin D receptor inhibits IgE production and that B cells can synthesize calcitriol from its precursor 25-hydroxyvitamin D3 (inactive precursor) [25(OH)D] upon antigenic stimulation. In this study, we address the impact of 25(OH)D on the development of type I sensitization and determine its role in allergen-specific immunotherapy. BALB/c mice were sensitized to OVA, under 25(OH)D-deficient or sufficient conditions. The humoral immune response over time was measured by ELISA. OVA-specific immunotherapy was established and studied in a murine model of allergic airway inflammation using lung histology, pulmonary cytokine expression analysis, and functional parameters in isolated and perfused mouse lungs. In 25(OH)D-deficient mice, OVA-specific IgE and IgG1 serum concentrations were increased compared with control mice. OVA-specific immunotherapy reduced the humoral immune reaction after OVA recall dose-dependently. Coadministration of 25(OH)D in the context of OVA-specific immunotherapy reduced the allergic airway inflammation and responsiveness upon OVA challenge. These findings were paralleled by reduced Th2 cytokine expression in the lungs. In conclusion, 25(OH)D deficiency promotes the development of type I sensitization and correction of its serum concentrations enhances the benefit of specific immunotherapy.

Topics: Animals; Calcifediol; Desensitization, Immunologic; Disease Models, Animal; Dose-Response Relationship, Immunologic; Drug Therapy, Combination; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Time Factors

The Th17 cytokines interleukin-17A (IL-17A), IL-17F, and IL-22 are critical for the lung immune response to a variety of bacterial pathogens, including Klebsiella pneumoniae. Th2 cytokine expression in the airways is a characteristic feature of asthma and allergic airway inflammation. The Th2 cytokines IL-4 and IL-13 diminish ex vivo and in vivo IL-17A protein expression by Th17 cells. To determine the effect of IL-4 and IL-13 on IL-17-dependent lung immune responses to acute bacterial infection, we developed a combined model in which allergic airway inflammation and lung IL-4 and IL-13 expression were induced by ovalbumin sensitization and challenge prior to acute lung infection with K. pneumoniae. We hypothesized that preexisting allergic airway inflammation decreases lung IL-17A expression and airway neutrophil recruitment in response to acute K. pneumoniae infection and thereby increases the lung K. pneumoniae burden. As hypothesized, we found that allergic airway inflammation decreased the number of K. pneumoniae-induced airway neutrophils and lung IL-17A, IL-17F, and IL-22 expression. Despite the marked reduction in postinfection airway neutrophilia and lung expression of Th17 cytokines, allergic airway inflammation significantly decreased the lung K. pneumoniae burden and postinfection mortality. We showed that the decreased lung K. pneumoniae burden was independent of IL-4, IL-5, and IL-17A and partially dependent on IL-13 and STAT6. Additionally, we demonstrated that the decreased lung K. pneumoniae burden associated with allergic airway inflammation was both neutrophil and CCL8 dependent. These findings suggest a novel role for CCL8 in lung antibacterial immunity against K. pneumoniae and suggest new mechanisms of orchestrating lung antibacterial immunity.

Topics: Animals; Chemokine CCL8; Eosinophils; Female; Hypersensitivity; Inflammation; Interleukins; Klebsiella Infections; Klebsiella pneumoniae; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Pneumonia, Bacterial

Curine is a bisbenzylisoquinoline alkaloid and the major constituent isolated from Chondrodendron platyphyllum, a plant that is used to treat inflammatory diseases in Brazilian folk medicine. This study investigates the effectiveness of curine on mast cell-dependent responses in mice.. To induce mast cell-dependent responses, Swiss mice were subcutaneously sensitized with ovalbumin (OVA-12 μg/mouse) and Al(OH)3 in a 0.9% NaCl solution. Fifteen days later, the animals were challenged with OVA through different pathways. Alternatively, the animals were injected with compound 48/80 or histamine, and several parameters, including anaphylaxis, itching, edema and inflammatory mediator production, were analyzed. Promethazine, cromoglycate, and verapamil were used as control drugs, and all of the treatments were performed 1h before the challenges.. Curine pre-treatment significantly inhibited the scratching behavior and the paw edema induced by either compound 48/80 or OVA, and this protective effect was comparable in magnitude with those associated with treatment with either cromoglycate or verapamil. In contrast, curine was a weak inhibitor of histamine-induced paw edema, which was completely inhibited by promethazine. Curine and verapamil significantly inhibited pleural protein extravasations and prostaglandin D2 (PGD2) and cysteinyl leukotrienes (CysLTs) production following allergen-induced pleurisy. Furthermore, like verapamil, curine inhibited the anaphylactic shock caused by either compound 48/80 or an allergen. In in vitro settings, these treatments also inhibited degranulation as well as PGD2 and CysLT production through IgE-dependent activation of the mast cell lineage RBL-2H3.. Curine significantly inhibited immediate allergic reactions through mechanisms more related to mast cell stabilization and activation inhibition than interference with the pro-inflammatory effects of mast cell products. These findings are in line with the hypothesis that the alkaloid curine may be beneficial for the treatment of allergic disorders.

Topics: Allergens; Animals; Anti-Allergic Agents; Brazil; Disease Models, Animal; Histamine; Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Isoquinolines; Male; Mast Cells; Medicine, Traditional; Menispermaceae; Mice; Ovalbumin

The prevalence of allergic diseases has doubled in developed countries in the past several decades. Cyclooxygenase (COX)-inhibiting drugs augmented allergic diseases in mice by increasing allergic sensitization and memory immune responses. However, whether COX inhibition can promote allergic airway diseases by inhibiting immune tolerance is not known.. To determine the role of the COX pathway and prostaglandin I2 (PGI2) signaling through the PGI2 receptor (IP) in aeroallergen-induced immune tolerance.. Wild-type (WT) BALB/c mice and IP knockout mice were aerosolized with ovalbumin (OVA) to induce immune tolerance prior to immune sensitization with an intraperitoneal injection of OVA/alum. The COX inhibitor indomethacin or vehicle was administered in drinking water to inhibit enzyme activity during the sensitization phase. Two weeks after sensitization, the mice were challenged with OVA aerosols. Mouse bronchoalveolar lavage fluid was harvested for cell counts and TH2 cytokine measurements.. WT mice treated with indomethacin had greater numbers of total cells, eosinophils, and lymphocytes, and increased IL-5 and IL-13 protein expression in BAL fluid compared to vehicle-treated mice. Similarly, IP knockout mice had augmented inflammation and TH2 cytokine responses compared to WT mice. In contrast, the PGI2 analog cicaprost attenuated the anti-tolerance effect of COX inhibition.. COX inhibition abrogated immune tolerance by suppressing PGI2 IP signaling, suggesting that PGI2 signaling promotes immune tolerance and that clinical use of COX-inhibiting drugs may increase the risk of developing allergic diseases.

Topics: Air Pollution; Allergens; Animals; Enzyme Inhibitors; Epoprostenol; Humans; Hypersensitivity; Immune Tolerance; Indomethacin; Mice; Mice, 129 Strain; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Prostaglandin-Endoperoxide Synthases; Receptors, Epoprostenol; Signal Transduction

Genetically modified (GM) foods are evaluated carefully for their ability to induce allergic disease. However, few studies have tested the capacity of a GM food to act as an adjuvant, i.e. influencing allergic responses to other unrelated allergens at acute onset and in individuals with pre-existing allergy. We sought to evaluate the effect of short-term feeding of GM Bacillus thuringiensis (Bt)-maize (MON810) on the initiation and relapse of allergic asthma in mice. BALB/c mice were provided a diet containing 33% GM or non-GM maize for up to 34 days either before ovalbumin (OVA)-induced experimental allergic asthma or disease relapse in mice with pre-existing allergy. We observed that GM-maize feeding did not affect OVA-induced eosinophilic airway and lung inflammation, mucus hypersecretion or OVA-specific antibody production at initiation or relapse of allergic asthma. There was no adjuvant effect upon GM-maize consumption on the onset or severity of allergic responses in a mouse model of allergic asthma.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Asthma; Bacillus thuringiensis; Female; Hypersensitivity; Lung; Mice, Inbred BALB C; Mucus; Ovalbumin; Plants, Genetically Modified; Pneumonia; Recurrence; Zea mays

Th17 cells are critical for the clearance of extracellular bacteria and fungi, but also contribute to the pathology of autoimmune diseases and allergic inflammation. After exposure to an appropriate cytokine environment, Th17 cells can acquire a Th1-like phenotype, but less is known about their ability to adopt Th2 and Th9 effector programs. To explore this in more detail, we used an IL-17F lineage tracer mouse strain that allows tracking of cells that formerly expressed IL-17F. In vitro-derived Th17 cells adopted signature cytokine and transcription factor expression when cultured under Th1-, Th2-, or Th9-polarizing conditions. In contrast, using two models of allergic airway disease, Th17 cells from the lungs of diseased mice did not adopt Th1, Th2, or Th9 effector programs, but remained stable IL-17 secretors. Although in vitro-derived Th17 cells expressed IL-4Rα, those induced in vivo during allergic airway disease did not, possibly rendering them unresponsive to IL-4-induced signals. However, in vitro-derived, Ag-specific Th17 cells transferred in vivo to OVA and aluminum hydroxide-sensitized mice also maintained IL-17 secretion and did not produce alternative cytokines upon subsequent OVA challenge. Thus, although Th17 cells can adopt new phenotypes in response to some inflammatory environments, our data suggest that in allergic inflammation, Th17 cells are comparatively stable and retain the potential to produce IL-17. This might reflect a cytokine environment that promotes Th17 stability, and allow a broader immune response at tissue barriers that are susceptible to allergic inflammation.

Topics: Aluminum Hydroxide; Animals; Asthma; Autoimmune Diseases; Cell Differentiation; Cell Lineage; Cytokines; Hypersensitivity; Interleukin-17; Lung; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Receptors, Cell Surface; Th1 Cells; Th17 Cells; Th2 Cells

Di-(2-ethylhexyl) phthalate (DEHP) has been considered as a widespread environmental persistent organic pollutant and its potential concern on human health is raised by previous studies. In particular, children are more likely to be exposed to DEHP through gastrointestinal route and consequently are more susceptible to DEHP hazards. Some reports have uncovered a positive association between DEHP exposure and an increased prevalence of allergic diseases in infants and juveniles. Allergy is a hypersensitive reaction rooted in imbalanced humoral immunity. T follicular helper cell (Tfh), an important CD4(+) Th cell subset, until recently has been identified as a key player in humoral immune response by modifying B cells functions. Tfh cells are therefore perceived as the therapeutic target of immune disorders. In the present study, focusing on the newly confirmed Tfh cells, we examined the effects of DEHP on humoral immunity and investigated the underlying mechanisms. Using ovalbumin (OVA) sensitization weanling mice model under the condition of gastrointestinal exposure to DEHP, we found that DEHP acted as an immunoadjuvant to augment OVA-specific IgE and IgG1 production, amplified germinal center formation in lymphoid nodule, as well as stimulated the expansion of CD4(+)CXCR5(+)ICOS(+)/CD4(+)CXCR5(+)PD-1(+) Tfh cells and CD19(+)CD138(+)GL7(+) plasma cells. Based on the results of immune adoptive transfusion, DEHP-related anaphylactic response was ascribed to Tfh cells hyperfunction directly. We further proved that DEHP gavage together with OVA sensitization adjuvantly promoted the synthesis of cytokines IL-21 and IL-4 and the expression of transcription factors Bcl-6 and c-Maf in Tfh cells. In conclusion, our study demonstrates that DEHP has adjuvant toxic effects on Tfh cells by synthesizing an excess of cytokines IL-21 and IL-4 via over-expression of transcription factors Bcl-6 and c-Maf, leading to an increasing secretion of allergy-related IgE and IgG1.

Topics: Administration, Oral; Adoptive Transfer; Animals; B-Lymphocytes; Cell Proliferation; Diethylhexyl Phthalate; Disease Models, Animal; DNA-Binding Proteins; Environmental Pollutants; Germinal Center; Hypersensitivity; Immunity, Humoral; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Interleukins; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-bcl-6; Proto-Oncogene Proteins c-maf; Risk Assessment; Risk Factors; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer

Our and others' previous studies have shown that Schistosoma japonicum (SJ) infection can inhibit allergic reactions. We recently reported that DCs played an important role in SJ infection-mediated inhibition of allergy, which was associated with enhanced IL-10 and T regulatory cell responses. Here, we further compared the role of CD8α(+) DC and CD8α(-) DC subsets for the inhibitory effect. We sorted CD8α(+) DC (SJCD8α(+) DC) and CD8α(-) DC (SJCD8α(-) DC) from SJ-infected mice and tested their ability to modulate allergic responses in vivo. The data showed that the adoptive transfer of SJCD8α(-) DC was much more efficient than SJCD8α(+) DC for the suppression of allergic airway eosinophilia, mucus overproduction, antigen-specific IgE responses, and Th2 cytokines (IL-4 and IL-5). More importantly, we found that the transfer of SJCD8α(-) DC, but not SJCD8α(+) DC, significantly increased IL-10 and TGF-β production following OVA exposure. As control, the transfer of DC subsets from naïve mice had no significant effect on allergic inflammation. In addition, SJCD8α-DC expressed significantly higher IL-10 but lower IL-12, CD80 and CD86 than SJCD8α(+) DC, fitting a tolerogenic phenotype. The results suggest that CD8α(-) DC is the predominant DC subset which is involved in the parasitic infection-mediated inhibition of allergic inflammation and possibly through enhancing immunomodulatory cytokine (IL-10 and TGF-β) production.

Topics: Adoptive Transfer; Animals; Antigens, Helminth; CD11b Antigen; Dendritic Cells; Female; Hypersensitivity; Interleukin-10; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Schistosomiasis japonica; T-Lymphocytes, Helper-Inducer; Transforming Growth Factor beta

The root bark of Morus alba L. (Mori Cortex Radicis; MCR) is traditionally used in Korean medicine for upper respiratory diseases. In this study, we investigated the antiasthmatic effect of kuwanon G isolated from MCR on ovalbumin (OVA)-induced allergic asthma in mice. Kuwanon G (1 and 10 mg/kg) was administered orally in mice once a day for 7 days during OVA airway challenge. We measured the levels of OVA-specific IgE and Th2 cytokines (IL-4, IL-5, and IL-13) in the sera or bronchoalveolar lavage (BAL) fluids and also counted the immune cells in BAL fluids. Histopathological changes in the lung tissues were analyzed. Kuwanon G significantly decreased the levels of OVA-specific IgE and IL-4, IL-5, and IL-13 in the sera and BAL fluids of asthma mice. Kuwanon G reduced the numbers of inflammatory cells in the BAL fluids of asthma mice. Furthermore, the pathological feature of lungs including infiltration of inflammatory cells, thickened epithelium of bronchioles, mucus, and collagen accumulation was inhibited by kuwanon G. These results indicate that kuwanon G prevents the pathological progression of allergic asthma through the inhibition of lung destruction by inflammation and immune stimulation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Flavonoids; Hypersensitivity; Immunoglobulin E; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Male; Mice, Inbred BALB C; Morus; Ovalbumin; Plant Bark; Plant Roots

This study investigates whether exposure to allergen elicits insulin resistance as a result of adipose tissue inflammation. Male C57BL/6 mice were challenged with ovalbumin (OVA) allergen for 12 weeks, and blood and adipose tissue samples were collected at 24h after the last challenge. Levels of adhesion molecules, fasting insulin, fasting glucose, and adipokines in the blood were analyzed, and fasting homeostasis model assessment was applied to determine insulin resistance (HOMA-IR). The expression of pro- and anti-inflammatory genes in dissected adipose tissues was analyzed by real-time RT-PCR. Our results showed that OVA exposure increased insulin resistance as well as resistin and E-selectin, but reduced adiponectin in the serum. Resistin level was significantly correlated with HOMA-IR. Moreover, in adipose tissues of OVA-challenged mice, the pro-inflammatory M1 genes were more abundant while the anti-inflammatory M2 genes were less than those of PBS-treated mice. The expressional changes of both M1 and M2 genes were significantly associated with serum levels of adiponectin, resistin, and E-selectin. Hematoxylin and eosin (HE) and immunohistochemistry (IHC) stain also showed that there was more obvious inflammation in OVA-challenged mice. In conclusion, the current study suggests the relationship between allergen-elicited adipose tissue inflammation and circulating inflammatory molecules, which are possible mediators for the development of insulin resistance. Therefore, we propose that allergen exposure might be one risk factor for insulin resistance.

Topics: Adiponectin; Adipose Tissue; Allergens; Animals; E-Selectin; Environmental Exposure; Humans; Hypersensitivity; Inflammation; Insulin Resistance; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Resistin

IL-22 is a Th17/Th22 cytokine that is increased in asthma. However, recent animal studies showed controversial findings in the effects of IL-22 in allergic asthma. To determine the role of IL-22 in ovalbumin-induced allergic inflammation we generated inducible lung-specific IL-22 transgenic mice. Transgenic IL-22 expression and signaling activity in the lung were determined. Ovalbumin (OVA)-induced pulmonary inflammation, immune responses, and airway hyperresponsiveness (AHR) were examined and compared between IL-22 transgenic mice and wild type controls. Following doxycycline (Dox) induction, IL-22 protein was readily detected in the large (CC10 promoter) and small (SPC promoter) airway epithelial cells. IL-22 signaling was evidenced by phosphorylated STAT3. After OVA sensitization and challenge, compared to wild type littermates, IL-22 transgenic mice showed decreased eosinophils in the bronchoalveolar lavage (BAL), and in lung tissue, decreased mucus metaplasia in the airways, and reduced AHR. Among the cytokines and chemokines examined, IL-13 levels were reduced in the BAL fluid as well as in lymphocytes from local draining lymph nodes of IL-22 transgenic mice. No effect was seen on the levels of serum total or OVA-specific IgE or IgG. These findings indicate that IL-22 has immune modulatory effects on pulmonary inflammatory responses in allergen-induced asthma.

Topics: Allergens; Animals; Chemokines; Gene Expression Regulation; Hypersensitivity; Immunomodulation; Interleukin-22; Interleukins; Mice; Mice, Transgenic; Organ Specificity; Ovalbumin; Pneumonia; STAT3 Transcription Factor

The human commensal microbiota interacts in a complex manner with the immune system, and the outcome of these interactions might depend on the immune status of the subject.. Previous studies have suggested a strong allergy-protective effect for Gammaproteobacteria. Here we analyze the skin microbiota, allergic sensitization (atopy), and immune function in a cohort of adolescents, as well as the influence of Acinetobacter species on immune responses in vitro and in vivo.. The skin microbiota of the study subjects was identified by using 16S rRNA sequencing. PBMCs were analyzed for baseline and allergen-stimulated mRNA expression. In in vitro assays human monocyte-derived dendritic cells and primary keratinocytes were incubated with Acinetobacter lwoffii. Finally, in in vivo experiments mice were injected intradermally with A lwoffii during the sensitization phase of the asthma protocol, followed by readout of inflammatory parameters.. In healthy subjects, but not in atopic ones, the relative abundance of Acinetobacter species was associated with the expression of anti-inflammatory molecules by PBMCs. Moreover, healthy subjects exhibited a robust balance between anti-inflammatory and TH1/TH2 gene expression, which was related to the composition of the skin microbiota. In cell assays and in a mouse model, Acinetobacter species induced strong TH1 and anti-inflammatory responses by immune cells and skin cells and protected against allergic sensitization and lung inflammation through the skin.. These results support the hypothesis that skin commensals play an important role in tuning the balance of TH1, TH2, and anti-inflammatory responses to environmental allergens.

Topics: Acinetobacter; Adolescent; Allergens; Animals; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Dendritic Cells; Gene Expression Profiling; Humans; Hypersensitivity; Keratinocytes; Leukocytes, Mononuclear; Mice; Microbiota; Ovalbumin; Pneumonia; RNA, Bacterial; RNA, Messenger; RNA, Ribosomal, 16S; Skin; Th1 Cells; Th2 Cells

Abnormality in mitochondria has been suggested to be associated with development of allergic airway disorders. In this study, to evaluate the relationship between mitochondrial reactive oxygen species (ROS) and NLRP3 inflammasome activation in allergic asthma, we used a newly developed mitochondrial ROS inhibitor, NecroX-5. NecroX-5 reduced the increase of mitochondrial ROS generation in airway inflammatory cells, as well as bronchial epithelial cells, NLRP3 inflammasome activation, the nuclear translocation of nuclear factor-κB, increased expression of various inflammatory mediators and pathophysiological features of allergic asthma in mice. Finally, blockade of IL-1β substantially reduced airway inflammation and hyperresponsiveness in the asthmatic mice. These findings suggest that mitochondrial ROS have a critical role in the pathogenesis of allergic airway inflammation through the modulation of NLRP3 inflammasome activation, providing a novel role of airway epithelial cells expressing NLRP3 inflammasome as an immune responder.

Topics: Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Carrier Proteins; Caspase 1; Cells, Cultured; DNA, Mitochondrial; Epithelial Cells; Heterocyclic Compounds, 4 or More Rings; Humans; Hypersensitivity; Inflammasomes; Inflammation; Interleukin-1beta; Intracellular Space; Lipopolysaccharides; Lung; Male; Mice; Middle Aged; Mitochondria; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Peroxidase; Reactive Oxygen Species; Sulfones; Trachea

The recruitment of CD4+CD25+Foxp3+T (Treg) cells is one of the most important mechanisms by which parasites down-regulate the immune system.. We compared the effects of Treg cells from Trichinella spiralis-infected mice and uninfected mice on experimental allergic airway inflammation in order to understand the functions of parasite-induced Treg cells. After four weeks of T. spiralis infection, we isolated Foxp3-GFP-expressing cells from transgenic mice using a cell sorter. We injected CD4+Foxp3+ cells from T. spiralis-infected [Inf(+)Foxp3+] or uninfected [Inf(-)Foxp3+] mice into the tail veins of C57BL/6 mice before the induction of inflammation or during inflammation. Inflammation was induced by ovalbumin (OVA)-alum sensitization and OVA challenge. The concentrations of the Th2-related cytokines IL-4, IL-5, and IL-13 in the bronchial alveolar lavage fluid and the levels of OVA-specific IgE and IgG1 in the serum were lower in mice that received intravenous application of Inf(+)Foxp3+ cells [IV(inf):+(+) group] than in control mice. Some features of allergic airway inflammation were ameliorated by the intravenous application of Inf(-)Foxp3+ cells [IV(inf):+(-) group], but the effects were less distinct than those observed in the IV(inf):+(+) group. We found that Inf(+)Foxp3+ cells migrated to inflammation sites in the lung and expressed higher levels of Treg-cell homing receptors (CCR5 and CCR9) and activation markers (Klrg1, Capg, GARP, Gzmb, OX40) than did Inf(-)Foxp3+ cells.. T. spiralis infection promotes the proliferation and functional activation of Treg cells. Parasite-induced Treg cells migrate to the inflammation site and suppress immune responses more effectively than non-parasite-induced Treg cells. The adoptive transfer of Inf(+)Foxp3+ cells is an effective method for the treatment and prevention of allergic airway diseases in mice and is a promising therapeutic approach for the treatment of allergic airway diseases.

Topics: Adoptive Transfer; Alum Compounds; Animals; Asthma; Cytokines; Female; Forkhead Transcription Factors; Hypersensitivity; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocytes, Regulatory; Trichinella spiralis; Trichinellosis

Oral tolerance (OT) is considered as a preventive and therapeutic strategy for treating asthma and allergic rhinitis (AR). We investigated the preventive effects of OT on allergic inflammation and remodeling in the upper and lower airways in a mouse model of allergy.. BALB/c mice were divided into four groups: control, allergy, low-dose OT, and high-dose OT. To induce OT, mice were fed ovalbumin (OVA) before sensitization with OVA/Al(OH)(3) at a dose of 1 mg for 6 days in low-dose OT group and a single dose of 25 mg in high-dose OT group. After sensitization followed by OVA challenge, nasal symptoms, interleukin (IL)-13, interferon (IFN)-gamma, IL-10, and transforming growth factor (TGF) beta-1 levels in nasal lavage (NAL) and bronchoalveolar lavage (BAL) fluids were measured, and OVA-specific IgE, IgG1, and IgG2a levels were measured in the serum. The airway hyperresponsiveness (AHR) was measured by enhanced pause. The goblet cell hyperplasia and the thickness of lamina propria were observed in the upper and lower airways.. In the allergy group, the allergic behavior scores, AHR, and OVA-specific IgE, IgG1, and IgG2a levels; inflammatory cells; IFN-gamma levels; and IL-13 levels in NAL/BAL fluids were elevated compared with the control group, low-dose OT group, and high-dose OT group. The allergy group had higher levels of IL-10 and TGF-beta-1 in BAL fluids when compared with the other groups. The goblet cell hyperplasia and the thickness of the lamina propria were attenuated in both OT groups compared with the allergy group.. OT may effectively prevent AHR, allergic inflammation, and airway remodeling in the upper and lower airways.

Topics: Administration, Oral; Airway Remodeling; Allergens; Animals; Cytokines; Disease Models, Animal; Female; Goblet Cells; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Immunotherapy; Inflammation; Mice; Mice, Inbred BALB C; Mouth; Mucous Membrane; Ovalbumin

Nucleotides released to the extracellular space stimulate purinergic receptors, and their effects are modulated by ectonucleotidases. The role of ATP in the allergic bronchospasm has been scantly studied.. We used several techniques (plethysmography, organ baths, confocal microscopy, RT-PCR, ATP measurement) to explore the role of nucleotides and ectonucleotidases in the allergic bronchospasm in guinea pigs.. While allergenic challenge with a low-dose ovalbumin (OVA) only produced a small bronchospasm (~2-fold the basal lung resistance), previous inhibition of ectonucleotidases by ARL-67156 greatly intensified this response (~11-fold the basal lung resistance, with 44% mortality). Bronchoalveolar lavage fluid obtained during this bronchospasm contained increased ATP concentration. This potentiation was abolished by antagonism of purinergic receptors (suramin+RB2) or TXA2 receptor (SQ29548), or by intratracheal apyrase. In tracheal rings and lung parenchyma strips, OVA caused a concentration-dependent contraction. Suramin+RB2 or levamisole produced a significant rightward displacement of this response, and ARL-67156 did not modify it. Platelets stimulated with OVA released ATP. Confocal images of nonsensitized tracheas showed slight fluorescence for P2Y6 receptors in epithelium and none for P2Y4 . Sensitized animals showed strong fluorescence to both receptors and to alkaline phosphatase in the airway epithelium. This correlated with a large increment in mRNA for P2Y4 and P2Y6 receptors in sensitized animals.. Nucleotides greatly potentiate the allergic bronchospasm when ectonucleotidases activity is diminished, and this effect is probably favored by the upregulation of P2Y4 and P2Y6 receptors in airway epithelium during sensitization. These results prompt for further research on these mechanisms in human asthma.

Topics: Adenosine Triphosphate; Alkaline Phosphatase; Animals; Blood Platelets; Bronchial Spasm; Bronchoalveolar Lavage Fluid; Dose-Response Relationship, Drug; Extracellular Space; Guinea Pigs; Hydrolysis; Hypersensitivity; Nucleotidases; Nucleotides; Ovalbumin; Receptors, Purinergic P2

A specific antigen-sensitized animal has antigen-specific immune cells that recognize the antigen. Therefore, an antigen-modified drug carrier would be recognized by the immune cells. When such a carrier encapsulates certain drugs, these drugs should be specifically delivered to the immune cells. To examine this strategy, ovalbumin (OVA) was used as model antigen, and mice were presensitized with 100 μg of OVA with Alum. For preparing OVA-modified liposomes (OVA-lipo), OVA was incubated with DSPE-PEG-NHS and resulting DSPE-PEG-OVA was inserted into liposomes. OVA-specific IgG was produced 6-fold higher by intravenous injection of OVA-lipo thrice (10 μg as OVA in each injection) in OVA-sensitized mice, than that by the injection of control liposomes, suggesting that OVA-lipo was recognized by the antigen-specific immune cells. Moreover, intra-splenic accumulation of OVA-lipo was observed in OVA-sensitized mice, but not in naive mice. To achieve the delivery of a drug to specific immune cells, OVA-lipo encapsulated low dose of doxorubicin (DOX) as a model drug (20 μg DOX/mouse, Ca. 1 mg/kg) was injected in the sensitized mice. The injection of OVA-lipo encapsulating DOX suppressed the production of IgE against OVA, suggesting that the specific delivery of the drug to immune cells responsible for OVA recognition was achieved and that these immune cells were removed by the drug treatment. This strategy would be useful for the fundamental treatment of allergy by the use of immunosuppressing agents.

Topics: Animals; Antibiotics, Antineoplastic; Antigens; Cholesterol; Doxorubicin; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Kidney; Liposomes; Liver; Lung; Mice; Mice, Inbred BALB C; Myocardium; Ovalbumin; Phosphatidylcholines; Phosphatidylethanolamines; Polyethylene Glycols; Spleen

The immunological adjuvant effect of silver nanoparticles (AgNPs) was investigated both in vitro and in vivo. The in vivo adjuvant effect of AgNPs was evaluated with model antigen ovalbumin (OVA) and bovine serum albumin (BSA) in mice by intraperitoneal and subcutaneous immunization. Serum antigen-specific IgG level significantly increased in AgNPs-treated mice comparing to the control group. AgNPs induced the increase of IgG1/IgG2a ratio and antigen-specific IgE, indicating that AgNPs elicited Th2-biased immune responses. By in vitro assay, the mechanism of adjuvant effect was explored. After 48h treatment with AgNPs, both the number of leukocytes and levels of cytokines TNF-α and IFN-γ in abdominal lavage fluid of mice increased. The expression of the major histocompatibility complex class II molecule on the surface of peritoneal macrophages significantly increased. AgNPs can be easily phagocytosed by peritoneal macrophages, while do not affect antigen uptake by the cell. We therefore conclude that AgNPs have significant adjuvant effect and the mechanism of this effect is mainly ascribed to the recruitment and activation of local leukocytes and especially macrophages. For the first time we found the remarkable adjuvant effect of AgNPs, and the result is beneficial for the future applications, especially in biomedicine.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Cytokinins; Female; Histocompatibility Antigens Class II; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Immunoglobulin Isotypes; Leukocytes; Macrophages, Peritoneal; Metal Nanoparticles; Mice; Mice, Inbred BALB C; Microscopy, Electron, Transmission; Ovalbumin; Particle Size; Serum Albumin, Bovine; Silver; Surface Properties

Long-term and unresolved airway inflammation and airway remodeling, characteristic features of chronic asthma, if not treated could lead to permanent structural changes in the airways. Aldose reductase (AR), an aldo-sugar and lipid aldehyde metabolizing enzyme, mediates allergen-induced airway inflammation in mice, but its role in the airway remodeling is not known. In the present study, we have examined the role of AR on airway remodeling using ovalbumin (OVA)-induced chronic asthma mouse model and cultured human primary airway epithelial cells (SAECs) and mouse lung fibroblasts (mLFs).. Airway remodeling in chronic asthma model was established in mice sensitized and challenged twice a week with OVA for 6 weeks. AR inhibitor, fidarestat, was administered orally in drinking water after first challenge. Inflammatory cells infiltration in the lungs and goblet cell metaplasia, airway thickening, collagen deposition and airway hyper-responsiveness (AHR) in response to increasing doses of methacholine were assessed. The TGFβ1-induced epithelial-mesenchymal transition (EMT) in SAECs and changes in mLFs were examined to investigate AR-mediated molecular mechanism(s) of airway remodeling.. In the OVA-exposed mice for 6 wks inflammatory cells infiltration, levels of inflammatory cytokines and chemokines, goblet cell metaplasia, collagen deposition and AHR were significantly decreased by treatment with AR inhibitor, fidarestat. Further, inhibition of AR prevented TGFβ1-induced altered expression of E-cadherin, Vimentin, Occludin, and MMP-2 in SAECs, and alpha-smooth muscle actin and fibronectin in mLFs. Further, in SAECs, AR inhibition prevented TGFβ1- induced activation of PI3K/AKT/GSK3β pathway but not the phosphorylation of Smad2/3.. Our results demonstrate that allergen-induced airway remodeling is mediated by AR and its inhibition blocks the progression of remodeling via inhibiting TGFβ1-induced Smad-independent and PI3K/AKT/GSK3β-dependent pathway.

Topics: Airway Remodeling; Aldehyde Reductase; Animals; Asthma; Biomarkers; Chronic Disease; Disease Models, Animal; Enzyme Inhibitors; Epithelial Cells; Epithelial-Mesenchymal Transition; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Humans; Hypersensitivity; Imidazolidines; Inflammation; Lung; Metaplasia; Mice; Mucus; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Signal Transduction; Smad Proteins; Transforming Growth Factor beta1

Up-regulation of arginase contributes to airways hyperresponsiveness (AHR) in asthma by reducing L-arginine bioavailability for the nitric oxide (NO) synthase isozymes. The product of arginase activity, L-ornithine, can be metabolized into polyamines by ornithine decarboxylase. We tested the hypothesis that increases in L-ornithine-derived polyamines contribute to AHR in mouse models of allergic airways inflammation. After measuring significantly increased polyamine levels in sputum samples from human subjects with asthma after allergen challenge, we used acute and subacute ovalbumin sensitization and challenge mouse models of allergic airways inflammation and naive mice to investigate the relationship of AHR to methacholine and polyamines in the lung. We found that spermine levels were elevated significantly in lungs from the acute model, which exhibits robust AHR, but not in the subacute murine model of asthma, which does not develop AHR. Intratracheal administration of spermine significantly augmented airways responsiveness to methacholine in both naive mice and mice with subacute airways inflammation, and reduced nitrite/nitrate levels in lung homogenates, suggesting that the AHR developed as a consequence of inhibition of constitutive NO production in the airways. Chronic inhibition of polyamine synthesis using an ornithine decarboxylase inhibitor significantly reduced polyamine levels, restored nitrite/nitrate levels to normal, and abrogated the AHR to methacholine in the acute model of allergic airways inflammation. We demonstrate that spermine increases airways responsiveness to methacholine, likely through inhibition of constitutive NO synthesis. Thus, inhibition of polyamine production may represent a new therapeutic target to treat airway obstruction in allergic asthma.

Topics: Adolescent; Adult; Animals; Asthma; Disease Models, Animal; Eflornithine; Female; Humans; Hypersensitivity; Inflammation; Lung; Male; Methacholine Chloride; Mice; Middle Aged; Nitrates; Nitric Oxide; Nitric Oxide Synthase; Ornithine; Ornithine Decarboxylase; Ornithine Decarboxylase Inhibitors; Ovalbumin; Polyamines; Spermine; Sputum; Young Adult

KCa3.1 has been suggested to be involved in regulating cell activation, proliferation, and migration in multiple cell types, including airway inflammatory and structural cells. However, the contributions of KCa3.1 to airway inflammation and remodeling and subsequent airway hyperresponsiveness (AHR) in allergic asthma remain to be explored. The main purpose of this study was to elucidate the roles of KCa3.1 and the potential therapeutic value of KCa3.1 blockers in chronic allergic asthma. Using real-time PCR, Western blotting, or immunohistochemical analyses, we explored the precise role of KCa3.1 in the bronchi of allergic mice and asthmatic human bronchial smooth muscle cells (BSMCs). We found that KCa3.1 mRNA and protein expression were elevated in the bronchi of allergic mice, and double labeling revealed that up-regulation occurred primarily in airway smooth muscle cells. Triarylmethane (TRAM)-34, a KCa3.1 blocker, dose-dependently inhibited the generation and maintenance of the ovalbumin-induced airway inflammation associated with increased Th2-type cytokines and decreased Th1-type cytokine, as well as subepithelial extracellular matrix deposition, goblet-cell hyperplasia, and AHR in a murine model of asthma. Moreover, the pharmacological blockade and gene silencing of KCa3.1, which was evidently elevated after mitogen stimulation, suppressed asthmatic human BSMC proliferation and migration, and arrested the cell cycle at the G0/G1 phase. In addition, the KCa3.1 activator 1-ethylbenzimidazolinone-induced membrane hyperpolarization and intracellular calcium increase in asthmatic human BSMCs were attenuated by TRAM-34. We demonstrate for the first time an important role for KCa3.1 in the pathogenesis of airway inflammation and remodeling in allergic asthma, and we suggest that KCa3.1 blockers may represent a promising therapeutic strategy for asthma.

Topics: Airway Remodeling; Animals; Asthma; Blotting, Western; Bronchi; Calcium Channel Agonists; Calcium Channel Blockers; Cell Membrane; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; Female; G1 Phase Cell Cycle Checkpoints; Gene Expression Regulation; Gene Silencing; Humans; Hypersensitivity; Immunohistochemistry; Inflammation; Intermediate-Conductance Calcium-Activated Potassium Channels; Membrane Potentials; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Pyrazoles; Real-Time Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Th2 Cells; Up-Regulation

The responses of airway rapidly adapting receptors (RARs) to ovalbumin challenge and histamine were investigated in guinea pigs which were sensitized with ovalbumin. Sensitization alone increased the basal RAR activity. Antigen challenge stimulated them. Histamine doses which caused a 50% increase in airway resistance (ED50) were reduced immediately and 24h after antigen challenge indicating respectively early and late onset airway hyperresponsiveness. At these doses, there was a greater stimulation of the RARs compared to controls. An increase in lipid peroxidation and a decrease in glutathione peroxidase were observed also. With oral intake of vitamins C and E, attenuations in the basal RAR activity, the responses of RARs to antigen challenge and the oxidative stress were observed. With an increase in ED50, the RAR response to histamine became similar as in control. It is concluded that by decreasing the RAR responses to allergen and histamine, antioxidants may reduce reflex bronchoconstriction occurring in asthmatics.

Topics: Action Potentials; Adaptation, Physiological; Airway Resistance; Animals; Ascorbic Acid; Bronchoconstriction; Erythrocyte Count; Glutathione Peroxidase; Guinea Pigs; Hemoglobins; Histamine; Histamine Agonists; Hypersensitivity; Ovalbumin; Oxidative Stress; Reactive Oxygen Species; Respiratory System; Spectrophotometry; Thiobarbituric Acid Reactive Substances; Vitamin E

Gamma radiation is used for several therapeutic indications such as cancers and autoimmune diseases. Low-dose whole-body γ irradiation has been shown to activate immune responses in several ways, however, the effect and mechanism of irradiation on allergic asthma remains poorly understood. This study investigated whether or not irradiation exacerbates allergic asthma responses and its potential mechanism. C57BL/6 mice were sensitized and challenged with ovalbumin (OVA) to induce asthma. The mice received whole-body irradiation once daily for 3 consecutive days with a dose of 0.667 Gy using (137)Cs γ rays 24 h before every OVA challenge. Repeated low-dose irradiation reduced OVA-specific IgE levels, the number of inflammatory cells including mast cells, goblet cell hyperplasia, collagen deposition, airway hyperresponsiveness, expression of inflammatory cytokines, CCL2/CCR2, as well as nuclear factor kappa B (NF-κB) and activator protein-1 activities. All of these factors were increased in BAL cells and lung tissue of OVA-challenged mice. Irradiation increased the number of Treg cells, expression of interleukin (IL)-10, IL-2 and IL-35 in BAL cells and lung tissue. Irradiation also increased Treg cell-expressed Foxp3 and IL-10 by NF-κB and RUNX1 in OVA-challenged mice. Furthermore, while Treg cell-expressing OX40 and IL-10 were enhanced in lung tissue or act-bone marrow-derived mast cells (BMMCs) with Treg cells, but BMMCs-expressing OX40L and TGF-β were decreased. The data suggest that irradiation enhances Foxp3(+)- and IL-10-producing Treg cells, which reduce OVA-induced allergic airway inflammation and tissue remodeling through the down-regulation of migration by the CCL2/CCR2 axis and activation of mast cells via OX40/OX40L in lung tissue of OVA-challenged mice.

Topics: Animals; Antibody Specificity; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Cell Size; Chemokines; Dose-Response Relationship, Radiation; Female; Forkhead Transcription Factors; Gamma Rays; Gene Expression Regulation; Histones; Hypersensitivity; Immunoglobulin E; Lung; Mast Cells; Mice; Mice, Inbred C57BL; NF-kappa B; Organ Specificity; Ovalbumin; OX40 Ligand; Protein Transport; Radiography; Receptors, Chemokine; Receptors, OX40; RNA, Messenger; Spleen; T-Lymphocytes, Regulatory; Transcription Factor AP-1

Volatile inhaled anesthetics exert a differential protective effect against bronchospasm development after cholinergic stimulation. However, their ability to inhibit the adverse respiratory consequences of an anaphylactic reaction after exposure to an allergen has not been characterized. We therefore compared the abilities of isoflurane, sevoflurane, and desflurane to prevent the lung constriction induced by an allergic reaction in a pediatric model of an anaphylactic reaction.. Low-frequency respiratory input impedance (Zrs) was measured in 4 groups of ovalbumin (OVA)-sensitized 5-week-old rabbit pups anesthetized with midazolam (group IV) and with inhaled isoflurane (group ISO), sevoflurane (group SEVO), or desflurane (group DES) at 1 minimum alveolar concentration. Zrs was measured under baseline conditions and after an anaphylactic lung response provoked by IV allergen injection (OVA 1 mg), during which the changes in airway resistance (Raw), tissue damping (G), and elastance obtained from Zrs were followed for 15 minutes.. Allergen provocation generated immediate severe bronchoconstriction, with no statistically significant difference in Raw increase among the groups in the first 3 minutes. Conversely, the inhalation of volatile anesthetics accelerated the recovery from the allergen-induced bronchoconstriction, particularly in group SEVO where the Raw was significantly lower than that in group IV 4 minutes after the allergen challenge. These changes were paralleled by significant elevations in G in all groups, with a significantly more pronounced deterioration in the animals in group DES. The anesthetic regimen did not statistically significantly affect the sustained increases in elastance after OVA injections.. Our results reveal the lack of potential of the commonly used volatile anesthetics to inhibit the most severe acute phase of the constrictor response to allergen after anaphylaxis in both the central airway and peripheral lung compartments. Inhalation of volatile anesthetics, particularly sevoflurane, promotes an earlier easing of the bronchospasm; this beneficial profile may be advantageous in children with atopic lung diseases.

Topics: Anesthetics, Inhalation; Animals; Blood Pressure; Bronchoconstriction; Female; Hypersensitivity; Male; Ovalbumin; Rabbits

We investigated the effects of orally administered probiotic bacteria (Lactobacillus species) as allergic immune modulators in ovalbumin (OVA)-sensitized mice. BALB/c mice were intraperitoneally injected with OVA twice at a 2-week interval for allergy sensitization. The mice were then orally administered Lactobacillus casei YIT9029 (L1), L. casei HY7201 (L2), L. brevis HY7401 (L3), or L. plantarum HY20301 (L4) every 2 days for 3 weeks. Total IgE levels significantly decreased in sera of L3-administered mice but increased in the other groups. OVA-specific IgE levels decreased slightly in sera of mice administered L1, L3, and L4 but increased significantly in L2-administered mice. In passive cutaneous anaphylaxis (PCA) using sera from administered mice, only the L3-administered group showed reaction inhibition. High expression of TLR-2 with interferon (IFN)-gamma stimulation on peripheral blood mononuclear cells occurred in L3- or L4-administered mice. Th1 cytokines, including IFN-gamma and interleukin (IL)- 12, increased in splenocytes of L3-administered mice; however, IL-4 decreased in L1- and L4-administered groups; IL-5 decreased in all experimental groups. IL-6 decreased in the L3-administered group; and IL-10 decreased in L1-, L2-, and L3-administered groups. L3 induced antiallergic effects by increasing Th1 cytokines, decreasing Th2 cytokines, and inhibiting the PCA reaction, whereas L2 administration increased allergic effects.

Topics: Administration, Oral; Animals; Anti-Allergic Agents; Cytokines; Female; Humans; Hypersensitivity; Immunoglobulin E; Immunologic Factors; Lactobacillus; Leukocytes, Mononuclear; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics

Asthma is a chronic inflammatory condition of the lower respiratory tract associated with airway hyperreactivity and mucus obstruction in which a majority of cases are due to an allergic response to environmental allergens. Glucocorticoids such as prednisone have been standard treatment for many inflammatory diseases for the past 60 years. However, despite their effectiveness, long-term treatment is often limited by adverse side effects believed to be caused by glucocorticoid receptor-mediated gene transcription. This has led to the pursuit of compounds that retain the anti-inflammatory properties yet lack the adverse side effects associated with traditional glucocorticoids. We have developed a novel series of steroidal analogues (VBP compounds) that have been previously shown to maintain anti-inflammatory properties such as NFκB-inhibition without inducing glucocorticoid receptor-mediated gene transcription. This study was undertaken to determine the effectiveness of the lead compound, VBP15, in a mouse model of allergic lung inflammation. We show that VBP15 is as effective as the traditional glucocorticoid, prednisolone, at reducing three major hallmarks of lung inflammation--NFκB activity, leukocyte degranulation, and pro-inflammatory cytokine release from human bronchial epithelial cells obtained from patients with asthma. Moreover, we found that VBP15 is capable of reducing inflammation of the lung in vivo to an extent similar to that of prednisone. We found that prednisolone--but not VBP15 shortens the tibia in mice upon a 5 week treatment regimen suggesting effective dissociation of side effects from efficacy. These findings suggest that VBP15 may represent a potent and safer alternative to traditional glucocorticoids in the treatment of asthma and other inflammatory diseases.

Topics: Animals; Asthma; Cell Degranulation; Cell Movement; Cytokines; Disease Models, Animal; Epithelial Cells; Female; Glucocorticoids; Humans; Hypersensitivity; Leukocytes; Lung; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Osteogenesis; Ovalbumin; Pneumonia; Pregnadienediols; Tibia

Allergic asthma is one of the most common chronic inflammatory diseases of airways. Severe asthma may lead to hospitalization and death. Sesame oil is a natural product with anti-inflammatory property. However, the effect of sesame oil on allergic asthma has never been studied.. We investigate the effect of sesame oil on pulmonary inflammation in allergic asthma model.. Allergic airway inflammation was induced by sensitizing with two doses of 10 mg ovalbumin (OVA) and then challenged with 1% OVA nebulizer exposure (1 h/day) for 3 days. Sesame oil (0.25, 0.5, or 1 mL/kg/day) was given orally 30 min before each challenge. Samples were collected 24 h after the last challenge.. Data showed that sesame oil inhibited pulmonary edema and decreased interleukin (IL)-1 β and IL-6 levels in bronchoalveolar lavage fluid in OVA-treated mice. Sesame oil also decreased pulmonary nitrite level, inducible nitric oxide synthase expression, and neutrophil infiltration induced by OVA. Further, sesame oil decreased serum IgE level in OVA-treated mice.. Sesame oil may attenuate pulmonary edema and bronchial neutrophilic inflammation by inhibiting systemic IgE level in allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Gene Expression Regulation; Humans; Hypersensitivity; Interleukin-1beta; Interleukin-6; Mice; Neutrophils; Nitric Oxide Synthase Type II; Ovalbumin; Pneumonia; Pulmonary Edema; Sesame Oil

Immunoglobulin E (IgE) is important for the development of allergic rhinitis (AR), though the contribution of IgE to the infiltration of eosinophils in the nasal mucosa has not been fully elucidated. In this study, antigen-induced sneezing and nasal eosinophil accumulation were comparatively investigated in anti-ovalbumin (OVA)-IgE transgenic (Tg) and wild-type (WT) mice.. Tg and OVA-immunized WT mice were intranasally challenged with OVA. Antigen-specific serum IgE level, sneezing and infiltration of eosinophil into the nasal cavity were then examined.. The level of serum OVA-specific IgE in Tg mice was significantly higher than that in antigen-immunized WT mice. Compared to saline challenge, intranasal challenge with OVA significantly induced sneezing in both Tg and immunized WT mice. However, antigen-induced nasal eosinophil infiltration was observed in immunized WT mice but not in Tg mice.. IgE-mediated responses might not play a crucial role in antigen-induced eosinophil infiltration in AR.

Topics: Animals; Antigens; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Transgenic; Nasal Mucosa; Ovalbumin; Rhinitis, Allergic, Perennial

It is well established that female sex hormones have a pivotal role in inflammation. For instance, our group has previously reported that estradiol has proinflammatory actions during allergic lung response in animal models. Based on these findings, we have decided to further investigate whether T regulatory cells are affected by female sex hormones absence after ovariectomy. We evaluated by flow cytometry the frequencies of CD4(+)Foxp3(+) T regulatory cells (Tregs) in central and peripheral lymphoid organs, such as the thymus, spleen and lymph nodes. Moreover, we have also used the murine model of allergic lung inflammation a to evaluate how female sex hormones would affect the immune response in vivo. To address that, ovariectomized or sham operated female Balb/c mice were sensitized or not with ovalbumin 7 and 14 days later and subsequently challenged twice by aerosolized ovalbumin on day 21. Besides the frequency of CD4(+)Foxp3(+) T regulatory cells, we also measured the cytokines IL-4, IL-5, IL-10, IL-13 and IL-17 in the bronchoalveolar lavage from lungs of ovalbumine challenged groups. Our results demonstrate that the absence of female sex hormones after ovariectomy is able to increase the frequency of Tregs in the periphery. As we did not observe differences in the thymus-derived natural occurring Tregs, our data may indicate expansion or conversion of peripheral adaptive Tregs. In accordance with Treg suppressive activity, ovariectomized and ovalbumine-sensitized and challenged animals had significantly reduced lung inflammation. This was observed after cytokine analysis of lung explants showing significant reduction of pro-inflammatory cytokines, such as IL-4, IL-5, IL-13 and IL-17, associated to increased amount of IL-10. In summary, our data clearly demonstrates that OVA sensitization 7 days after ovariectomy culminates in reduced lung inflammation, which may be directly correlated with the expansion of Tregs in the periphery and further higher IL-10 secretion in the lungs.

Topics: Animals; Bronchoalveolar Lavage Fluid; CD4 Antigens; Cytokines; Female; Flow Cytometry; Forkhead Transcription Factors; Gonadal Steroid Hormones; Hypersensitivity; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Ovariectomy; Spleen; T-Lymphocytes, Regulatory

One feature of allergic asthma, the EAR (early allergic reaction), is not present in the commonly used mouse models. We therefore investigated the mediators involved in EAR in a guinea-pig in vivo model of allergic airway inflammation. Animals were sensitized using a single OVA (ovalbumin)/alum injection and challenged with aerosolized OVA on day 14. On day 15, airway resistance was assessed after challenge with OVA or MCh (methacholine) using the forced oscillation technique, and lung tissue was prepared for histology. The contribution of mast cell mediators was investigated using inhibitors of the main mast cell mediators [histamine (pyrilamine) and CysLTs (cysteinyl-leukotrienes) (montelukast) and prostanoids (indomethacin)]. OVA-sensitized and challenged animals demonstrated AHR (airway hyper-responsiveness) to MCh, and lung tissue eosinophilic inflammation. Antigen challenge induced a strong EAR in the sensitized animals. Treatment with a single compound, or indomethacin together with pyrilamine or montelukast, did not reduce the antigen-induced airway resistance. In contrast, dual treatment with pyrilamine together with montelukast, or triple inhibitor treatment, attenuated approximately 70% of the EAR. We conclude that, as in humans, the guinea-pig allergic inflammation model exhibits both EAR and AHR, supporting its suitability for in vivo identification of mast cell mediators that contribute to the development of asthma. Moreover, the known mast cell mediators histamine and leukotrienes were major contributors of the EAR. The data also lend further support to the concept that combination therapy with selective inhibitors of key mediators could improve asthma management.

Topics: Acetates; Animals; Asthma; Bronchial Hyperreactivity; Constriction, Pathologic; Cyclopropanes; Disease Models, Animal; Guinea Pigs; Histamine Antagonists; Hypersensitivity; Indomethacin; Leukotriene Antagonists; Lung; Mast Cells; Ovalbumin; Prostaglandin Antagonists; Pyrilamine; Quinolines; Sulfides

Rapid desensitization transiently prevents severe allergic reactions, allowing administration of life-saving therapies in previously sensitized patients. However, the mechanisms underlying successful rapid desensitization are not fully understood.. We sought to investigate whether the mast cell (MC) is an important target of rapid desensitization in mice sensitized to exhibit IgE-dependent passive systemic anaphylaxis in vivo and to investigate the antigen specificity and underlying mechanisms of rapid desensitization in our mouse model.. C57BL/6 mice (in vivo) or primary isolated C57BL/6 mouse peritoneal mast cells (PMCs; in vitro) were passively sensitized with antigen-specific anti-2,4-dinitrophenyl IgE, anti-ovalbumin IgE, or both. MCs were exposed over a short period of time to increasing amounts of antigen (2,4-dinitrophenyl-human serum albumin or ovalbumin) in the presence of extracellular calcium in vitro or by means of intravenous administration to sensitized mice in vivo before challenging the mice with or exposing the PMCs to optimal amounts of specific or irrelevant antigen.. Rapidly exposing mice or PMCs to progressively increasing amounts of specific antigen inhibited the development of antigen-induced hypothermia in sensitized mice in vivo and inhibited antigen-induced PMC degranulation and prostaglandin D2 synthesis in vitro. Such MC hyporesponsiveness was induced antigen-specifically and was associated with a significant reduction in antigen-specific IgE levels on MC surfaces.. Rapidly exposing MCs to progressively increasing amounts of antigen can both enhance the internalization of antigen-specific IgE on the MC surface and also desensitize these cells in an antigen-specific manner in vivo and in vitro.

Topics: 2,4-Dinitrophenol; Anaphylaxis; Animals; Antibodies, Anti-Idiotypic; Antibody Specificity; Antigens; Desensitization, Immunologic; Humans; Hypersensitivity; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred C57BL; Ovalbumin; Time Factors

Asthma is a chronic inflammatory disease of the small airways, with airway hyperresponsiveness (AHR) and inflammation as hallmarks. Recent studies suggest a role for arginase in asthma pathogenesis, possibly because arginine is the substrate for both arginase and NO synthase and because NO modulates bronchial tone and inflammation. Our objective was to investigate the importance of increased pulmonary arginase 1 expression on methacholine-induced AHR and lung inflammation in a mouse model of allergic asthma. Arginase 1 expression in the lung was ablated by crossing Arg1(fl/fl) with Tie2Cre(tg/-) mice. Mice were sensitized and then challenged with ovalbumin. Lung function was measured with the Flexivent. Adaptive changes in gene expression, chemokine and cytokine secretion, and lung histology were quantified with quantitative PCR, ELISA, and immunohistochemistry. Arg1 deficiency did not affect the allergic response in lungs and large-airway resistance, but it improved peripheral lung function (tissue elastance and resistance) and attenuated adaptive increases in mRNA expression of arginine-catabolizing enzymes Arg2 and Nos2, arginine transporters Slc7a1 and Slc7a7, chemokines Ccl2 and Ccl11, cytokines Tnfa and Ifng, mucus-associated epithelial markers Clca3 and Muc5ac, and lung content of IL-13 and CCL11. However, expression of Il4, Il5, Il10, and Il13 mRNA; lung content of IL-4, IL-5, IL-10, TNF-α, and IFN-γ protein; and lung pathology were not affected. Correlation analysis showed that Arg1 ablation disturbed the coordinated pulmonary response to ovalbumin challenges, suggesting arginine (metabolite) dependence of this response. Arg1 ablation in the lung improved peripheral lung function and affected arginine metabolism but had little effect on airway inflammation.

Topics: Airway Resistance; Animals; Arginase; Asthma; Blotting, Western; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Chemokines; Cytokines; Dendritic Cells; Female; Gene Expression Profiling; Hypersensitivity; Immunoenzyme Techniques; Lung; Macrophages; Male; Methacholine Chloride; Mice; Mice, Inbred C57BL; Mice, Knockout; Myeloid Cells; Ovalbumin; Pneumonia; Real-Time Polymerase Chain Reaction; Respiratory System; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

To constitute an animal model of laryngeal allergy and evaluate the laryngeal effects of inhaled corticosteroids and ß2-agonists on the laryngeal mucosa in an allergic rat model.. Prospective randomized.. The Experimental Medical Research Institute (DETAE) at Istanbul University.. Wistar Albino rats (n = 32) were sensitized with ovalbumin. Unsensitized rats (n = 8) served as controls. The rats were exposed to aerosolized ovalbumin (1%). On days 28 through 42, every 2 days preceeding ovalbumin exposure, rats were further exposed to aerosolized phosphate buffered saline (n = 8), fluticasone propionate (n = 8), salbutamol (n = 8), and combined salbutamol+fluticasone propionate (n = 8). Inflammatory cell infiltration was graded semi-quantitatively. The quantitative data included mast cell count and degranulation. Ultrathin sections were investigated under transmission electron microscope.. The simultaneous and pairwise comparison of groups (Kruskal-Wallis) revealed statistically significant difference among groups at supraglottic level (critical P < .05, <.01) and no difference at glottic level. In ovalbumin+phosphate buffered saline exposed rats, the light microscopy of supraglottic mucosa revealed regular epithelium with severe inflammatory cell infiltration and increased mast cell count. Electron microscopy revealed increased mast cell degranulation. Increased inflammatory cell infiltration was detected along with reduced mast cell count among fluticasone propionate treated rats. Mild inflammatory cell infiltration was encountered in combined salbutamol+fluticasone propionate treated rats.. This study supported the presence of localized allergic reaction in the supraglottic laryngeal mucosa through the observation of increased mast cell number and degranulation. It was also shown that inhaled corticosteroids increase inflammation whereas combined inhaled corticosteroids and ß2-agonists minimize allergic and inflammatory reactions in supraglottic laryngeal mucosa providing a safer therapeutic option.

Topics: Administration, Inhalation; Adrenal Cortex Hormones; Albuterol; Androstadienes; Animals; Disease Models, Animal; Fluticasone; Hypersensitivity; Laryngeal Mucosa; Mast Cells; Ovalbumin; Prospective Studies; Random Allocation; Rats; Rats, Wistar

This study was carried out to determine the effect of allergic inflammation on the airway response to increasing airway temperature. Our results showed the following: 1) In Brown-Norway rats actively sensitized by ovalbumin (Ova), isocapnic hyperventilation with humidified warm air (HWA) for 2 min raised tracheal temperature (Ttr) from 33.4 ± 0.6°C to 40.6 ± 0.1°C, which induced an immediate and sustained (>10 min) increase in total pulmonary resistance (Rl) from 0.128 ± 0.004 to 0.212 ± 0.013 cmH2O·ml(-1)·s (n = 6, P < 0.01). In sharp contrast, the HWA challenge caused the same increase in Ttr but did not generate any increase in Rl in control rats. 2) The increase in Rl in sensitized rats was reproducible when the same HWA challenge was repeated 60-90 min later. 3) This bronchoconstrictive effect was temperature dependent: a slightly smaller increase in peak Ttr (39.6 ± 0.2°C) generated a significant but smaller increase in Rl in sensitized rats. 4) The HWA-induced bronchoconstriction was not generated by the humidity delivered by the HWA challenge alone, because the same water content delivered by saline aerosol at room temperature had no effect. 5) The HWA-evoked increase in Rl in sensitized rats was not blocked by atropine but was completely prevented by pretreatment either with a combination of neurokinin (NK)-1 and NK-2 antagonists or with formoterol, a β2 agonist, before the HWA challenge. This study showed that increasing airway temperature evoked a pronounced and reversible increase in airway resistance in sensitized rats and that tachykinins released from the vagal bronchopulmonary C-fiber endings were primarily responsible.

Topics: 2,4-Dichlorophenoxyacetic Acid; Aerosols; Airway Resistance; Animals; Atropine; Body Temperature; Bronchoconstriction; Humidity; Hypersensitivity; Hyperventilation; Inflammation; Male; Ovalbumin; Rats; Rats, Inbred BN; Tachykinins; Trachea; Vagus Nerve

Neuroimmune semaphorin 4D (Sema4D) was found to be expressed and function in the nervous and immune systems. In the immune system, Sema4D is constitutively expressed on T cells and regulates T cell priming. In addition, it displays a stimulatory function on macrophages, DC, NK cells, and neutrophils. As all these cells are deeply involved in asthma pathology, we hypothesized that Sema4D plays a critical non-redundant regulatory role in allergic airway response. To test our hypothesis, we exposed Sema4D(-/-) and WT mice to OVA injections and challenges in the well-defined mouse model of OVA-induced experimental asthma. We observed a significant decrease in eosinophilic airway infiltration in allergen-treated Sema4D(-/-) mice relative to WT mice. This reduced allergic inflammatory response was associated with decreased BAL IL-5, IL-13, TGFβ1, IL-6, and IL-17A levels. In addition, T cell proliferation in OVA₃₂₃₋₃₃₉-restimulated Sema4D(-/-) cell cultures was downregulated. We also found increased Treg numbers in spleens of Sema4D(-/-) mice. However, airway hyperreactivity (AHR) to methacholine challenges was not affected by Sema4D deficiency in either acute or chronic experimental disease setting. Surprisingly, lung DC number and activation were not affected by Sema4D deficiency. These data provide a new insight into Sema4D biology and define Sema4D as an important regulator of Th2-driven lung pathophysiology and as a potential target for a combinatory disease immunotherapy.

Topics: Animals; Antigens, CD; Asthma; Bronchoalveolar Lavage Fluid; Cell Proliferation; Cells, Cultured; Cytokines; Flow Cytometry; Humans; Hypersensitivity; Lung; Lymphocyte Activation; Lymphocytes; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pneumonia; Pulmonary Eosinophilia; Semaphorins; Th1 Cells; Th17 Cells; Th2 Cells

The importance of the lung parenchyma in the pathophysiology of asthma has previously been demonstrated. Considering that nitric oxide synthases (NOS) and arginases compete for the same substrate, it is worthwhile to elucidate the effects of complex NOS-arginase dysfunction in the pathophysiology of asthma, particularly, related to distal lung tissue. We evaluated the effects of arginase and iNOS inhibition on distal lung mechanics and oxidative stress pathway activation in a model of chronic pulmonary allergic inflammation in guinea pigs.. Guinea pigs were exposed to repeated ovalbumin inhalations (twice a week for 4 weeks). The animals received 1400 W (an iNOS-specific inhibitor) for 4 days beginning at the last inhalation. Afterwards, the animals were anesthetized and exsanguinated; then, a slice of the distal lung was evaluated by oscillatory mechanics, and an arginase inhibitor (nor-NOHA) or vehicle was infused in a Krebs solution bath. Tissue resistance (Rt) and elastance (Et) were assessed before and after ovalbumin challenge (0.1%), and lung strips were submitted to histopathological studies.. Ovalbumin-exposed animals presented an increase in the maximal Rt and Et responses after antigen challenge (p<0.001), in the number of iNOS positive cells (p<0.001) and in the expression of arginase 2, 8-isoprostane and NF-kB (p<0.001) in distal lung tissue. The 1400 W administration reduced all these responses (p<0.001) in alveolar septa. Ovalbumin-exposed animals that received nor-NOHA had a reduction of Rt, Et after antigen challenge, iNOS positive cells and 8-isoprostane and NF-kB (p<0.001) in lung tissue. The activity of arginase 2 was reduced only in the groups treated with nor-NOHA (p <0.05). There was a reduction of 8-isoprostane expression in OVA-NOR-W compared to OVA-NOR (p<0.001).. In this experimental model, increased arginase content and iNOS-positive cells were associated with the constriction of distal lung parenchyma. This functional alteration may be due to a high expression of 8-isoprostane, which had a procontractile effect. The mechanism involved in this response is likely related to the modulation of NF-kB expression, which contributed to the activation of the arginase and iNOS pathways. The association of both inhibitors potentiated the reduction of 8-isoprostane expression in this animal model.

Topics: Administration, Inhalation; Animals; Arginase; Chronic Disease; Dinoprost; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Lung; Male; NF-kappa B; Nitric Oxide Synthase Type II; Ovalbumin; Oxidative Stress; Pneumonia; Respiratory Mechanics

Previous studies have indicated that colitis increases intestinal permeability to food antigens. This condition also generates an immunoreactive milieu in the gut, which may exacerbate or counteract allergy reactions. This, along with the fact that both colitis and allergy are being codiagnosed more frequently, means the scientific interest on the immune relation between these pathologies is increasing. We evaluated the immune response to an internalized food antigen that was initiated during a concomitant active intestinal inflammatory response.. An ovalbumin (OVA)-induced immune response was analyzed in healthy mice and in mice suffering from colitis induced by the administration of dinitrofluorobenzene/dinitrosulfonic acid (DNFB/DNS) at the moment of OVA challenge. The OVA-induced clinical score and allergy response both in plasma and in splenocyte cultures from these animals were compared.. Although no differences were observed in the allergy clinical score, the concomitant active colitis led to an increase in the immune response to OVA antigen, as shown by increased spleen size and OVA-induced splenocyte proliferation, exacerbated expression of total and OVA-specific IgG1 levels, increased colonic IL-4 expression and OVA-induced IL-4 and IL-5 cytokine expression in spleen cells.. Our results indicate that animals with active colitis undergo an exacerbated immune response to an internalized antigen. This finding could be relevant for the allergy management of patients presenting simultaneously with chronic colitis.

Topics: Animals; Antigens; Colitis; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lymphocyte Activation; Lymphocytes; Mice; Ovalbumin; Spleen

There is a growing concern for the possible health impact of nanoparticles. The main objective of this study was to investigate the allergy-promoting capacity of four different carbon nanofiber (CNF) samples in an injection and an airway mouse model of allergy. Secondly, the potency of the CNF was compared to the previously reported allergy-promoting capacity of carbon nanotubes (CNT) in the airway model. Ultrafine carbon black particles (ufCBP) were used as a positive control. Particles were given together with the allergen ovalbumin (OVA) either by subcutaneous injection into the footpad or intranasally to BALB/cA mice. After allergen booster, OVA-specific IgE, IgG1, and IgG2a in serum were measured. In the airway model, inflammation was determined as influx of inflammatory cells (eosinophils, neutrophils, lymphocytes, and macrophages) and by mediators (MCP-1 and TNF-α present in bronchoalveolar fluid (BALF)). CNF and CNT both increased OVA-specific IgE levels in the two models, but in the airway model, the CNT gave a significantly stronger IgE response than the CNF. Furthermore, the CNT and not the CNF promoted eosinophil lung inflammation. Our data therefore suggest that nanotube-associated properties are particularly potent in promoting allergic responses.

Topics: Adjuvants, Immunologic; Animals; Carbon; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Nanofibers; Nanotubes, Carbon; Ovalbumin; Tumor Necrosis Factor-alpha

A reproducible method to inhibit allergic immune responses is accomplished with hi-dose Ag sensitization, via intraperitoneal (IP) injection. However, the role of CD4+ CD25+ FoxP3+ T regulatory cells (Treg) in this process is unknown, as is whether such modulation extends to ocular allergy. We therefore determined herein whether hi-dose sensitization modulates ocular allergy, and whether CD4+ CD25+ FoxP3+ Treg are involved. C57BL/6 mice were IP sensitized via low-dose (100 µg) versus hi-dose (1000 µg) ovalbumin (OVA), in aluminum hydroxide (1 mg) and pertussis-toxin (300 ng). Other mice received anti-CD25 Ab (PC61) to ablate Treg during sensitization. In another experiment, Treg from hi-dose sensitized mice were adoptively transferred into low-dose sensitized mice. Once daily OVA challenges were administered. Clinical signs, IgE, T cell cytokines, and eosinophils were assessed. Data revealed that hi-dose, but not low-dose, sensitization led to allergy modulation, indicated by decreased clinical signs, serum IgE levels, Th2 recall responses, and eosinophil recruitment. T cells from hi-dose sensitized mice showed a robust increase in TGF-b production, and Treg from these mice were able to efficiently suppress effector T cell proliferation in vitro. In addition, in vivo Treg ablation in hi-dose sensitized mice revoked allergy modulation. Lastly, Treg from hi-dose sensitized mice were able to adoptively transfer allergy modulation to their low-dose sensitized counterparts. Collectively, these findings indicate that modulation to hi-dose sensitization, which is extended to ocular allergy, occurs in a Treg-dependent manner. In addition, our data suggest that hi-dose sensitization may henceforth facilitate the further examination of CD4+ CD25+ FoxP3+ Treg in allergic disease.

Topics: Animals; Antigens; Cell Proliferation; Cytokines; Eosinophils; Forkhead Transcription Factors; Hypersensitivity; Immunization; Immunoglobulin E; Interleukin-2 Receptor alpha Subunit; Male; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells; Transforming Growth Factor beta

Stimulatory IgG receptors (FcγRs) on bone marrow-derived cells contribute to the pathogenesis of several autoimmune and inflammatory disorders. Monoclonal antibodies that block FcγRs might suppress these diseases, but they can induce anaphylaxis.. We wanted to determine whether a rapid desensitization approach can safely suppress IgG/FcγR-mediated anaphylaxis.. Mice were injected with serially increasing doses of 2.4G2, a rat mAb that blocks the inhibitory FcγR, FcγRIIb, and the stimulatory receptor, FcγRIII. Rectal temperature was used to detect the development of anaphylaxis. Passive and active IgG-mediated anaphylaxis were evaluated in mice that had been rapidly desensitized with 2.4G2 or mock-desensitized in mice in which monocyte/macrophages, basophils, or neutrophils had been depleted or desensitized and in mice in which FcγRI, FcγRIII, and/or FcγRIV had been deleted or blocked.. Rapid desensitization with 2.4G2 prevented 2.4G2-induced shock and completely suppressed IgG-mediated anaphylaxis. Rapid desensitization of ovalbumin-sensitized mice with 2.4G2 was safer and more effective than rapid desensitization with ovalbumin. 2.4G2 treatment completely blocked FcγRIII and removed most FcγRI and FcγRIV from nucleated peripheral blood cells. Because IgG(2a)-mediated anaphylaxis was partially FcγRI and FcγRIV dependent, the effects of 2.4G2 on FcγRI and FcγRIV were probably crucial for its complete inhibition of IgG(2a)-mediated anaphylaxis. IgG(2a)-mediated anaphylaxis was partially inhibited by depletion or desensitization of monocyte/macrophages, basophils, or neutrophils.. IgG-mediated anaphylaxis can be induced by ligation of FcγRI, FcγRIII, or FcγRIV on monocycte/macrophages, basophils, or neutrophils and can be safely suppressed by rapid desensitization with anti-FcγRII/RIII mAb. A similar approach may safely suppress other FcγR-dependent immunopathology.

Topics: Anaphylaxis; Animals; Antibodies, Blocking; Antibodies, Monoclonal; Basophils; Body Temperature; Desensitization, Immunologic; Disease Models, Animal; Female; Hypersensitivity; Macrophages; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Rats; Receptors, IgG

Compare the effects of montelukast or dexamethasone in distal lung parenchyma and airway walls of guinea pigs (GP) with chronic allergic inflammation.. GP have inhaled ovalbumin (OVA group-2x/week/4weeks). After the 4th inhalation, GP were treated with montelukast or dexamethasone. After 72 hours of the 7th inhalation, GP were anesthetised, and lungs were removed and submitted to histopathological evaluation.. Montelukast and dexamethasone treatments reduced the number of eosinophils in airway wall and distal lung parenchyma compared to OVA group (P < 0.05). On distal parenchyma, both treatments were effective in reducing RANTES, NF- κ B, and fibronectin positive cells compared to OVA group (P < 0.001). Montelukast was more effective in reducing eotaxin positive cells on distal parenchyma compared to dexamethasone treatment (P < 0.001), while there was a more expressive reduction of IGF-I positive cells in OVA-D group (P < 0.001). On airway walls, montelukast and dexamethasone were effective in reducing IGF-I, RANTES, and fibronectin positive cells compared to OVA group (P < 0.05). Dexamethasone was more effective in reducing the number of eotaxin and NF- κ B positive cells than Montelukast (P < 0.05).. In this animal model, both treatments were effective in modulating allergic inflammation and remodeling distal lung parenchyma and airway wall, contributing to a better control of the inflammatory response.

Topics: Acetates; Administration, Inhalation; Animals; Chronic Disease; Cyclopropanes; Dexamethasone; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Inflammation; Leukotriene Antagonists; Lung; Ovalbumin; Quinolines; Sulfides

Allergic airway inflammation is attenuated by oral tolerization (oral exposure to allergen, followed by conventional sensitization and challenge with homologous antigen), which decreases airway allergen challenge-induced eosinophilic infiltration of the lungs and bone marrow eosinophilia. We examined its effects on bone marrow eosinophil and neutrophil production. Mice of wild type (BP-2, BALB/c, and C57BL/6) and mutant strains (lacking iNOS or CD95L) were given ovalbumin (OVA) or water (vehicle) orally and subsequently sensitized and challenged with OVA (OVA/OVA/OVA and H2O/OVA/OVA groups, resp.). Anti-OVA IgG and IgE, bone marrow eosinophil and neutrophil numbers, and eosinophil and neutrophil production ex vivo were evaluated. T lymphocytes from OVA/OVA/OVA or control H2O/OVA/OVA donors were transferred into naïve syngeneic recipients, which were subsequently sensitized/challenged with OVA. Alternatively, T lymphocytes were cocultured with bone marrow eosinophil precursors from histocompatible sensitized/challenged mice. OVA/OVA/OVA mice of the BP-2 and BALB/c strains showed, relative to H2O/OVA/OVA controls, significantly decreased bone marrow eosinophil counts and ex vivo eosinopoiesis/neutropoiesis. Full effectiveness in vivo required sequential oral/subcutaneous/intranasal exposures to the same allergen. Transfer of splenic T lymphocytes from OVA/OVA/OVA donors to naive recipients prevented bone marrow eosinophilia and eosinopoiesis in response to recipient sensitization/challenge and supressed eosinopoiesis upon coculture with syngeneic bone marrow precursors from sensitized/challenged donors.

Topics: Administration, Oral; Allergens; Animals; Bone Marrow Cells; Eosinophils; Hematopoiesis; Humans; Hypersensitivity; Immunity, Innate; Inflammation; Lung; Mice; Ovalbumin; T-Lymphocytes

To study the biomedical functions of dioscorins isolated from various species of Dioscorea , we investigated their antiallergic potential using an OVA-induced allergy mouse model. All the dioscorins suppressed allergic reactions by decreasing the serum IgE and histamine levels. The serum IFN-γ and IgG2a levels increased in all the dioscorin-treated mice. The spleen cells from the dioscorin-treated mice also exhibited an up-regulation of IFN-γ secretion in response to ConA stimulation. Although dioscorins did not affect the IgG1 levels, the IL-5 levels decreased to basal levels in mice treated with dioscorins of D. alata or D. japonica and in most of the lymphoid cells of the dioscorin-treated mice in response to ConA stimulation. The decrease of IgE and histamine levels was concomitant with an increase in IFN-γ and IgG2a levels and with a decrease in IL-5 levels, suggesting that dioscorins suppressed the OVA-induced allergic reactions, possibly through modulating an imbalanced Th1/Th2 immune response.

Topics: Animals; Anti-Allergic Agents; Dioscorea; Histamine; Hypersensitivity; Immunoglobulin G; Interferon-gamma; Interleukin-5; Male; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Proteins; Plant Tubers; Spleen; Th1 Cells; Th2 Cells

To clarify the relationship between selenium supplementation and type I allergic reaction, we investigated the effect of seleno-L-methionine (SeMet) supplementation on the active cutaneous anaphylaxis (ACA) reaction and cytokine production in splenocytes. Female BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), and SeMet was administered orally for 2 weeks followed by a challenge with OVA to induce an ACA reaction. SeMet supplementation suppressed the ACA reaction in a dose-dependent manner. Plasma OVA-specific immunoglobulin E (IgE) level was strongly inhibited in SeMet-supplemented mice compared with control mice. The mRNA expression levels of the T helper 2 (Th2) cytokines interleukin (IL)-4 and IL-13 in the spleen of SeMet-supplemented mice were lower than those in control mice. The mRNA expression level of a Th1 cytokine, interferon (IFN)-γ, in the spleen of SeMet-supplemented mice was higher than that in control mice. Splenocytes restimulated with OVA in vitro from SeMet-supplemented mice produced lower amounts of IL-4 and IL-13 than those of control mice and higher amounts of IFN-γ than those from the control mice. These results suggest that oral SeMet supplementation suppresses OVA-induced ACA reaction by lowered Th2 cytokine production and augmenting Th1 cytokine production.

Topics: Anaphylaxis; Animals; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Liver; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Messenger; Selenomethionine; Skin Tests; Spleen

Vascular endothelial growth factor (VEGF) is supposed to contribute to the pathogenesis of allergic airway disease. VEGF expression is regulated by a variety of stimuli such as nitric oxide, growth factors, and hypoxia-inducible factor-1 alpha (HIF-1α). Recently, inhibition of the mammalian target of rapamycin (mTOR) has been shown to alleviate cardinal asthmatic features, including airway hyperresponsiveness, eosinophilic inflammation, and increased vascular permeability in asthma models. Based on these observations, we have investigated whether mTOR is associated with HIF-1α-mediated VEGF expression in allergic asthma. In studies with the mTOR inhibitor rapamycin, we have elucidated the stimulatory role of a mTOR-HIF-1α-VEGF axis in allergic response. Next, the mechanisms by which mTOR is activated to modulate this response have been evaluated. mTOR is known to be regulated by phosphoinositide 3-kinase (PI3K)/Akt or protein kinase C-delta (PKC δ) in various cell types. Consistent with these, our results have revealed that suppression of PKC δ by rottlerin leads to the inhibition of PI3K/Akt activity and the subsequent blockade of a mTOR-HIF-1α-VEGF module, thereby attenuating typical asthmatic attack in a murine model. Thus, the present data indicate that PKC δ is necessary for the modulation of the PI3K/Akt/mTOR signaling cascade, resulting in a tight regulation of HIF-1α activity and VEGF expression. In conclusion, PKC δ may represent a valuable target for innovative therapeutic treatment of allergic airway disease.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Female; Gene Expression Regulation; Humans; Hypersensitivity; Hypoxia-Inducible Factor 1, alpha Subunit; Lung; Mice; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoproteins; Protein Kinase C-delta; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; RNA Interference; RNA, Small Interfering; Signal Transduction; Th2 Cells; TOR Serine-Threonine Kinases; Vascular Endothelial Growth Factor A

Topics: Adjuvants, Immunologic; Animals; Humans; Hypersensitivity; Ligands; Ovalbumin; Recombinant Fusion Proteins; Toll-Like Receptors; Vaccines

Calcium release-activated calcium channels (CRAC) play unambiguous role in secretory functions of mast cells, T cells, and eosinophils. Less knowledge exists about the role of CRAC, widely distributed in airway smooth muscle (ASM) cells, in airway contractility. The presented study seeks to determine the possible participation of CRAC in ASM-based inflammatory airway disorders in guinea pigs. The acute and long-term administration (14 days) of the CRAC antagonist 3-fluoropyridine-4-carboxylic acid was used to examine the ASM contractility and associated reflexes in the guinea pig model of allergic airway inflammation by the following methods: (i) evaluation of specific airway resistance in vivo; (ii) evaluation of the contractile response of isolated ASM strips in vitro; and (iii) citric acid-induced cough reflex; (iv) measurement of exhaled NO levels (E(NO)). Allergic airway inflammation was induced by repetitive exposure of guinea pigs to ovalbumin (10(-6) M). The CRAC antagonist administered in a single dose to guinea pigs with confirmed allergic inflammation significantly reduced the cough response and the airway resistance, which corresponded with the findings in vitro. Long-term application of the CRAC antagonist had more strongly expressed effects. The results confirm the role of CRAC in the pathophysiology of experimental animal asthma and have a potential meaning for anti-asthma therapy.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Calcium; Calcium Channel Blockers; Calcium Channels; Cough; Endoplasmic Reticulum; Eosinophils; Guinea Pigs; Humans; Hypersensitivity; Isonicotinic Acids; Male; Mast Cells; Muscle Contraction; Myocytes, Smooth Muscle; Nitric Oxide; Ovalbumin; Respiratory System; T-Lymphocytes

Fish oil supplementation during pregnancy has been associated with lower levels of cord blood IL-13, suggesting that the administration of n-3 fatty acids may attenuate the development of allergic disease. The present study aimed to investigate the mechanism by which n-3 fatty acid administration influences the production of IL-13. Pregnant BALB/c mice were fed nutritionally complete high-fat diets (15 %, w/w) with an n-3 fatty acid-enriched (DHA 1 %, w/w) or control diet (0 % DHA) immediately following delivery. Pups were exposed during suckling and weaned to the maternal diet for the remainder of the study. The production of IL-13, IL-4, IL-10 and interferon-γ from the splenocytes of ovalbumin (ova)-sensitised animals was assessed following in vitro ova stimulation or unstimulated conditions. Human T helper type 2 (Th2) cells were mitogen-stimulated in the presence or absence of DHA (10 μM) and assessed for IL-13 and IL-4 expression using intracellular flow cytometry. The influence on transcriptional activation was studied using a human IL-13 promoter reporter construct and electromobility shift assay. Ova-activated splenocytes from DHA-fed mice produced less IL-13 (57.2 (se 21.7) pg/ml) and IL-4 (7.33 (SE 3.4) pg/ml) compared with cells from the animals fed the control diet (161.5 (SE 45.0), P< 0.05; 33.2 (SE 11.8), P< 0.05). In vitro, DHA inhibited the expression of IL-13 protein from human Th2 cells as well as transcriptional activation and binding of the transcription factors cyclic AMP response element binding and activating transcription factor 2 to the human IL-13 promoter. These data indicate the potential of n-3 fatty acids to attenuate IL-13 expression, and suggest that they may subsequently reduce allergic sensitisation and the development of allergic disease.

Topics: Animals; Diet, High-Fat; Docosahexaenoic Acids; Fatty Acids, Omega-3; Female; Humans; Hypersensitivity; Interferon-gamma; Interleukin-10; Interleukin-13; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Promoter Regions, Genetic; Spleen; Th2 Cells; Transcription, Genetic

Cissampelos sympodialis Eichl. (Menispermaceae) root infusion is used in Northeast Brazil to treat allergic asthma. We have previously shown that oral use of the plant extract reduces eosinophil infiltration into the lung of ovalbumin (OVA)- sensitized mice. However, drugs taken by inhalation route to treat asthma achieve better outcomes. Thereby, in this study, we evaluated the inhaled C. sympodialis alcoholic extract as a therapeutic treatment in OVA-sensitized BALB/c mice. The parameters which were analyzed consisted of leukocyte recruitment to the airway cavity, tissue remodeling and cell profile. The inhaled extract inhibited mainly eosinophil recruitment to the pleural cavity, bronchoalveolar lavage and peripheral blood. This treatment reduced the OVA-specific IgE serum titer and leukocyte infiltration in the peribronchiolar and pulmonary perivascular areas as well as mucus production. In addition, we also tested isolated alkaloids from the plant extract. The flow cytometric analysis showed that methylwarifteine (MW) and, mainly, the inhaled extract reduced the number of CD3+T cells and eosinophil-like cells. Therefore, inhaled C. sympodialis extract and MW lead to down-regulation of inflammatory cell infiltration with remarkable decrease in the number of T cells in an experimental model of respiratory allergy, suggesting that the plant can be delivered via inhalation route to treat allergic asthma.

Topics: Administration, Inhalation; Alkaloids; Animals; Benzylisoquinolines; CD3 Complex; Cissampelos; Down-Regulation; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Pleurisy; Rats; Rats, Wistar; T-Lymphocytes

A recent epidemiological study has revealed the positive association between atopy morbidity in children and phthalate esters, environmental chemicals in house dust. Nonetheless, experimental and molecular evidences regarding the correlation between phthalates and allergic response/pathophysiology are not fully investigated. Among phthalate esters, di-(2-ethylhexyl) phthalate (DEHP) has been widely used for flexible polyvinyl chloride products including vinyl flooring and wall covering. In the present study, we examined the effects of exposure to DEHP on allergen (ovalbumin: OVA) -induced peritonitis in ICR mice. Repeated administration of OVA via intraperitoneal route induced peritoneal inflammation characterized by infiltration of granulocytes (neutrophils and eosinophils) into the cavity. DEHP synergistically exaggerated the OVA-related neutrophilic inflammation. Furthermore, DEHP + OVA profoundly amplified OVA-elicited inflammation- and allergy-related molecules such as interleukin-5, eotaxin, and keratinocyte-derived chemoattractant production/release in the peritoneal cavity. Taken together, DEHP aggravated OVA-related peritoneal inflammation, which is concomitant with local enhanced production/release of inflammation- and allergy-related molecules.

Topics: Animals; Ascitic Fluid; Chemokines; Cytokines; Diethylhexyl Phthalate; Eosinophils; Hypersensitivity; Inflammation; Interleukin-5; Keratinocytes; Male; Mice; Mice, Inbred ICR; Neutrophils; Ovalbumin; Peritonitis

During the last decade, there has been a remarkable and unexplained increase in the prevalence of asthma. These studies were conducted to investigate the role of dermal exposure to triclosan, an endocrine-disrupting compound, on the hypersensitivity response to ovalbumin (OVA) in a murine model of asthma. Triclosan has had widespread use in the general population as an antibacterial and antifungal agent and is commonly found in consumer products such as soaps, deodorants, toothpastes, shaving creams, mouthwashes, and cleaning supplies. For these studies, BALB/c mice were exposed dermally to concentrations of triclosan ranging from 0.75 to 3% (0.375-1.5mg/mouse/day) for 28 consecutive days. Concordantly, mice were ip injected with OVA (0.9 µg) and aluminum hydroxide (0.5mg) on days 1 and 10 and challenged with OVA (125 µg) by pharyngeal aspiration on days 19 and 27. Compared with the animals exposed to OVA alone, increased spleen weights, OVA-specific IgE, interleukin-13 cytokine levels, and numbers of lung eosinophils were demonstrated when mice were coexposed to OVA and triclosan. Statistically significant increases in OVA-specific and nonspecific airway hyperreactivity were observed for all triclosan coexposed groups compared with the vehicle and OVA controls. In these studies, exposure to triclosan alone was not demonstrated to be allergenic; however, coexposure with a known allergen resulted in enhancement of the hypersensitivity response to that allergen, suggesting that triclosan exposure may augment the allergic responses to other environmental allergens.

Topics: Administration, Cutaneous; Animals; Anti-Infective Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Interleukin-13; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Messenger; Triclosan

Allergic asthma is a chronic airway disorder characterized by airway hyperresponsiveness to allergens, chronic airway inflammation, airway edema, increased mucus secretion, excess production of Th2 cytokines, and eosinophil accumulation in the lungs. Ursolic acid is known for its pharmacological effects, such as its anti-tumor, anti-inflammatory and antimicrobial activities. To investigate the anti-asthmatic effects and mechanism of ursolic acid, we studied the development of pulmonary eosinophilic inflammation and enhanced pause (Penh) in a mouse model of allergic asthma. In this study, BALB/c mice were systemically sensitized to ovalbumin followed by intratracheal, intraperitoneal, and aerosol allergen challenges. We investigated the effect of ursolic acid and Cyclosporin A (CsA) on Penh, pulmonary eosinophilic infiltration, various immune cell phenotypes, Th2 cytokines, IL-17 production, and ovalbumin specific IgE production in a mouse model of asthma. In BALB/c mice, ursolic acid had suppressed eosinophil infiltration, allergic airway inflammation, and Penh, which occurred by suppressing the production of IL-5, IL-13, IL-17, and ovalbumin-specific IgE by blocking the GATA-3 and STAT6 pathways. Our data suggest the therapeutic mechanism of ursolic acid in asthma is based on reductions of Th2 cytokines (IL-5 and IL-13), ovalbumin-specific IgE production, and eosinophil infiltration via the Th2-GATA-3, STAT6, and IL-17-NF-κB pathways.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cell Line; Cyclosporine; Disease Models, Animal; Down-Regulation; Eosinophils; Female; GATA3 Transcription Factor; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-17; Interleukin-5; Interleukins; Lung; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; RNA, Messenger; STAT6 Transcription Factor; Th2 Cells; Triterpenes; Ursolic Acid

Human rhinovirus (HRV) infections lead to exacerbations of lower airways disease in asthmatic patients but not in healthy individuals. However, underlying mechanisms remain to be completely elucidated. We hypothesized that the Th2-driven allergic environment enhances HRV-induced CC chemokine production, leading to asthma exacerbations. Ovalbumin (OVA)-sensitized and -challenged mice inoculated with HRV showed significant increases in the expression of lung CC chemokine ligand (CCL)-2/monocyte chemotactic protein (MCP)-1, CCL4/macrophage inflammatory protein (MIP)-1β, CCL7/MCP-3, CCL19/MIP-3β, and CCL20/MIP3α compared with mice treated with OVA alone. Inhibition of CCL2 with neutralizing antibody significantly attenuated HRV-induced airways inflammation and hyperresponsiveness in OVA-treated mice. Immunohistochemical stains showed colocalization of CCL2 with HRV in epithelial cells and CD68-positive macrophages, and flow cytometry showed increased CCL2(+), CD11b(+) cells in the lungs of OVA-treated, HRV-infected mice. Compared with lung macrophages from naïve mice, macrophages from OVA-exposed mice expressed significantly more CCL2 in response to HRV infection ex vivo. Pretreatment of mouse lung macrophages and BEAS-2B human bronchial epithelial cells with interleukin (IL)-4 and IL-13 increased HRV-induced CCL2 expression, and mouse lung macrophages from IL-4 receptor knockout mice showed reduced CCL2 expression in response to HRV, suggesting that exposure to these Th2 cytokines plays a role in the altered HRV response. Finally, bronchoalveolar macrophages from children with asthma elaborated more CCL2 upon ex vivo exposure to HRV than cells from nonasthmatic patients. We conclude that CCL2 production by epithelial cells and macrophages contributes to HRV-induced airway hyperresponsiveness and inflammation in a mouse model of allergic airways disease and may play a role in HRV-induced asthma exacerbations.

Topics: Animals; Antibodies, Neutralizing; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Chemokine CCL19; Chemokine CCL2; Chemokine CCL20; Chemokine CCL4; Chemokine CCL7; Child; Disease Models, Animal; Epithelial Cells; Gene Expression; Humans; Hypersensitivity; Inflammation; Interleukin-13; Interleukin-4; Lung; Macrophages; Mice; Ovalbumin; Rhinovirus; Th2 Cells

Asthma is characterized by airway hyperresponsiveness and remodeling. Pravastatin and atorvastatin are used clinically as cholesterol-lowering agents but also exhibit anti-inflammatory and immunomodulating properties.. To investigate the therapeutic effect of oral statins on airway hyperresponsiveness and allergic reaction.. BALB/c mice received intraperitoneal sensitization and aerosol inhalation with ovalbumin consequently. One week after ovalbumin aerosol challenge, pravastatin, atorvastatin, or phosphate-buffered saline were given by intragastric gavage daily for 2 weeks. Airway hyperresponsiveness, serum allergen specific antibody levels, cytokine production by splenocytes, and bronchoalveolar lavage fluid were examined.. Both pravastatin and atorvastatin effectively reduced airway hyperresponsiveness. Pravastatin effectively suppressed both T(H)1- and T(H)2-mediated antibody responses, reducing serum specific IgE, IgG, IgG1, and IgG2a levels. Pravastatin also effectively reduced interleukin (IL) 4, IL-5, and interferon γ production but significantly enhanced IL-10 levels in splenocytes and BALF. Similarly, atorvastatin effectively attenuated production of specific IgE, IgG1, and IgG2a antibodies. It also significantly attenuated IL-4, interferon γ, and increased IL-10 concentration in bronchoalveolar lavage fluid and splenocytes.. Oral administration of pravastatin or atorvastatin not only was able to inhibit T(H)1 inflammatory responses but also had therapeutic effects on airway hyperresponsiveness and T(H)2 allergic responses. These results seem to suggest that these drugs have potential as a nonimmunosuppressive therapy for asthma and allergic diseases.

Topics: Administration, Oral; Animals; Asthma; Atorvastatin; Bronchial Hyperreactivity; Cytokines; Female; Heptanoic Acids; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hypersensitivity; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; Pravastatin; Pyrroles; Rats, Sprague-Dawley; Th1 Cells

Asthma is a complex disease involving genetic and environment interactions. Atopy is a strong risk factor for asthma. The airway epithelium not only forms a physical barrier but also provides immune defense against harmful materials. To explore the effects of airway epithelium on asthma, we hypothesized that environmental injuries could act on bronchial epithelial cells and damage the physical barrier, which might facilitate allergens to stimulate immunoreactions and play an important role in the pathogenesis of asthma.. Thirty eight-week-old male Wistar rats were randomly divided into five groups with six rats in each group: control group, asthma group, ovalbumin (OVA) + OVA group, lipopolysaccharide (LPS) group and LPS + OVA group. In the control group, 0.9% saline was injected intraperitoneally on day 1. Fourteen days later, the rats were exposed to aerosolized 0.9% saline. In the asthma group, the rats were sensitized with an injection of 10 mg of OVA, followed by an aerosolized 2% OVA challenge 14 days later. The OVA + OVA group was sensitized by an inhalation 2% OVA, 20 minutes a day, from day 1 to day 7, and then OVA challenged in the same way as the asthma group. In the LPS group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with saline as in the control group. While in the LPS + OVA group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with OVA as in the asthma group. The expression of interleukin (IL)-4, interferon-gamma (IFN-γ) and thymic stromal lymphopoietin (TSLP) in the lungs was detected by reverse transcription polymerase chain reaction (RT-PCR) and the pulmonary pathological changes were also observed. The level of IL-4, IFN-γ and IgE in plasma was detected by enzyme-linked immunosorbent assay (ELISA). Bronchoalveolar lavage fluid (BALF) was collected to conduct differential cell counts. Flow cytometry analysis was also used to count Th1 and Th2 cells.. The pathological changes in the LPS + OVA group were similar to the asthma group, while in other groups, the pathological changes were not obvious. The ratio of lymphocytes in BALF, IL-4/IFN-γ in plasma and the expression of the TSLP and IL-4 in the asthma and LPS + OVA groups were higher than in the control group and the OVA + OVA group (P < 0.05). The level of IgE was higher in the asthma, LPS and LPS + OVA groups than in the control group and the OVA + OVA group (P < 0.05). By flow cytometry analysis, the Th1/Th2 ratio was lower in the LPS + OVA and asthma groups than in other groups (P < 0.05).. The experiment results show that the injury to the bronchial epithelial layer may be the initial event of allergic responses. This finding implies that a rational approach to therapeutics would be to increase the resistance of the airways to environmental injuries rather than concentrating on suppressing inflammation.

Topics: Animals; Bronchi; Cell Count; Cytokines; Disease Models, Animal; Epithelial Cells; Hypersensitivity; Immunoglobulin E; Interferon-gamma; Interleukin-4; Lipopolysaccharides; Male; Ovalbumin; Rats; Rats, Wistar; Thymic Stromal Lymphopoietin

Although CD23-dependent transcytosis of IgE and IgE-derived immune complexes across respiratory epithelial cells is likely to play a pivotal role in the initiation and development of airway allergic inflammation, there is currently a lack of physiological support for this phenomena to suggest that the targeting of CD23 could be used as a means of therapeutic intervention. The present study was designed to detect the CD23 expression in the nasal mucosa of allergic rhinitis (AR) murine model by immunohistochemistry and western blotting, and to investigate whether intranasal anti-CD23 treatment could inhibit allergen-induced upper airway inflammation in the AR model. This is the first report to show that CD23 was constitutively expressed in murine nasal epithelial cells, and its expression was significantly up-regulated in the AR murine model. In vivo, the up-regulation of CD23 expression was correlated with increased serum IL-4 levels. Following intranasal anti-CD23 treatment, nasal symptoms were alleviated and histopathologic examination showed a significant decrease in eosinophilic infiltration. Meanwhile, ELISA analysis showed levels of serum leukotriene C4 (LTC4), eosinophil cation protein (ECP), ovalbumin (OVA)-specific IgE and IL-4 also significantly decreased, as were LTC4 and OVA-specific IgE in the nasal lavage fluid. Furthermore, Western blotting analysis showed that ECP expression in the nasal mucosa was down-regulated. Finally, flow cytometric analysis revealed anti-CD23 treatment inhibited Th2 cell responses. These results indicate that intranasal anti-CD23 treatment can reduce allergic responses in a murine model of allergic rhinitis.

Topics: Administration, Intranasal; Allergens; Animals; Budesonide; Disease Models, Animal; Down-Regulation; Eosinophil Cationic Protein; Eosinophils; Epithelial Cells; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Leukotriene C4; Mice; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Random Allocation; Receptors, IgE; Rhinitis, Allergic; Rhinitis, Allergic, Perennial; Th2 Cells; Up-Regulation

Glucocorticoids are widely regarded as the most effective treatment for asthma. However, the direct impact of glucocorticoids on the innate immune system and antibacterial host defense during asthma remain unclear. Understanding the mechanisms underlying this process is critical to the clinical application of glucocorticoids for asthma therapy. After sensitization and challenge with ovalbumin (OVA), BALB/c mice were treated with inhaled budesonide and infected with Pseudomonas aeruginosa (P. aeruginosa). The number of viable bacteria in enflamed lungs was evaluated, and levels of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum were measured. A lung epithelial cell line was pretreated with budesonide. Levels of cathelicidin-related antimicrobial peptide (CRAMP) were measured by immunohistochemistry and western blot analysis. Intracellular bacteria were observed in lung epithelial cells.. Inhaled budesonide enhanced lung infection in allergic mice exposed to P. aeruginosa and increased the number of viable bacteria in lung tissue. Higher levels of IL-4 and lower levels of IFN-γ were observed in the serum. Budesonide decreased the expression of CRAMP, increased the number of internalized P. aeruginosa in OVA-challenged mice and in lung epithelial cell lines. These data indicate that inhaled budesonide can suppress pulmonary antibacterial host defense by down-regulating CRAMP in allergic inflammation mice and in cells in vitro.. Inhaled budesonide suppressed pulmonary antibacterial host defense in an asthmatic mouse model and in lung epithelium cells in vitro. This effect was dependent on the down-regulation of CRAMP.

Topics: Animals; Anti-Bacterial Agents; Antimicrobial Cationic Peptides; Budesonide; Cathelicidins; Colony Count, Microbial; Down-Regulation; Epithelial Cells; Host-Pathogen Interactions; Hypersensitivity; Immunosuppression Therapy; Inflammation; Interferon-gamma; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pseudomonas aeruginosa; Pseudomonas Infections

Development of allergic asthma is a complex process involving immune, neuronal and tissue cells. In the lung, Clara cells represent a major part of the "immunomodulatory barrier" of the airway epithelium. To understand the contribution of these cells to the inflammatory outcome of asthma, disease development was assessed using an adjuvant-free ovalbumin model. Mice were sensitised with subcutaneous injections of 10 μg endotoxin-free ovalbumin in conjunction with naphthalene-induced Clara cell depletion. Clara epithelial cell depletion in the lung strongly reduced eosinophil influx, which correlated with decreased eotaxin levels and, moreover, diminished the T-helper cell type 2 inflammatory response, including interleukin (IL)-4, IL-5 and IL-13. In contrast, airway hyperresponsiveness was increased. Further investigation revealed Clara cells as the principal source of eotaxin in the lung. These findings are the first to show that Clara airway epithelial cells substantially contribute to the infiltration of eotaxin-responsive CCR3+ immune cells and augment the allergic immune response in the lung. The present study identifies Clara cells as a potential therapeutic target in inflammatory lung diseases such as allergic asthma.

Topics: Allergens; Animals; Asthma; Chemokine CCL11; Cytokines; Eosinophils; Female; Hypersensitivity; Mice; Mice, Inbred BALB C; Naphthalenes; Ovalbumin; Receptors, CCR3; Respiratory Mucosa; T-Lymphocytes

Allergic diseases are featured by an increased production of IgE due to an imbalance in the immune response towards a Th2 profile. In this work, the ability of Enterococcus faecalis CECT7121 to regulate this Th2-exaggerated response in a murine model of ovalbumin (OVA)-induced allergy was studied.. BALB/c mice intragastrically inoculated with E. faecalis CECT7121 before and during a subcutaneous immunization protocol with OVA were studied in comparison with an immunized control group. The allergen-specific immune response (IgE, IgG, IgG1 and IgG2a) was assessed. The proliferative activity of memory splenocytes and the levels of IL-4, IL-5, IL-13, IL-10, IL-12 and IFN-γ were also determined.. Upon treatment with E. faecalis CECT7121 the following effects were observed: (1) a decrease in specific IgE levels, (2) an increase in anti-OVA IgG2a levels, (3) the levels of anti-OVA IgG and IgG1 remained unaltered, (4) a reduction in the proliferation rate of memory cells, (5) a decrease in the levels of the Th2 cytokines IL-4, IL-5 and IL-13, and (6) the secretion of IL-10, IL-12 and IFN-γ remained unchanged. Moreover, the incubation of human basophils with non-viable E. faecalis CECT7121 together with an allergen preparation induced the release of β-hexosaminidase at levels that were lower than control reactions and similar i.g. the spontaneous release.. In this model, the i.g. administration of E. faecalis CECT7121 hampers the establishment of the OVA-induced allergic immune response, suggesting that this strain could be useful for the treatment of IgE-mediated allergic diseases.

Topics: Allergens; Animals; Antigens, Bacterial; Cytokines; Desensitization, Immunologic; Enterococcus faecalis; Female; Hypersensitivity; Immunity, Cellular; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Skin Tests

Dendritic cells (DCs) have the ability to present antigen and play a critical role in the induction of the acquired immune response. Skin DCs uptake antigen and subsequently migrate to regional draining lymph nodes (LNs), where they activate naive T cells. Here we show that the water/glycerol channel protein aquaporin 7 (AQP7) is expressed on epidermal and dermal DCs and involved in the initiation of primary immune responses. AQP7-deficient DCs showed a decreased cellular uptake of low-molecular-mass compounds (fluorescein isothiocyanate and Lucifer yellow) and high-molecular-mass substances (ovalbumin and dextran), suggesting that AQP7 is involved in antigen uptake. AQP7-deficient DCs also exhibited reduced chemokine-dependent cell migration in comparison to wild-type DCs. Consistent with these in vitro results, AQP7-deficient mice demonstrated a reduced accumulation of antigen-retaining DCs in the LNs after antigen application to the skin, which could be attributed to decreased antigen uptake and migration. Coincidentally, AQP7-deficient mice had impaired antigen-induced sensitization in a contact hypersensitivity model. These observations suggested that AQP7 in skin DCs is primarily involved in antigen uptake and in the subsequent migration of DCs and is responsible for antigen presentation and the promotion of downstream immune responses.

Topics: Animals; Antigens; Aquaporins; Cell Movement; Cells, Cultured; Chemotaxis; Dendritic Cells; Dermatitis, Contact; Dermis; Disease Models, Animal; Epidermal Cells; Epidermis; Glycerol; Haptens; Hypersensitivity; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pinocytosis; Water

Epidemiological studies suggest that stress has an impact on asthmatic exacerbations. We evaluated if repeated stress, induced by forced swimming, modulates lung mechanics, distal airway inflammation and extracellular matrix remodeling in guinea pigs with chronic allergic inflammation.. Guinea pigs were submitted to 7 ovalbumin or saline aerosols (1-5 mg/ml during 4 weeks; OVA and SAL groups). Twenty-four hours after the 4th inhalation, guinea pigs were submitted to the stress protocol 5 times a week during 2 weeks (SAL-S and OVA-S groups). Seventy-two hours after the 7th inhalation, guinea pigs were anesthetized and mechanically ventilated. Resistance and elastance of the respiratory system were obtained at baseline and after ovalbumin challenge. Lungs were removed, and inflammatory and extracellular matrix remodeling of distal airways was assessed by morphometry. Adrenals were removed and weighed.. The relative adrenal weight was greater in stressed guinea pigs compared to non-stressed animals (p < 0.001). Repeated stress increased the percent elastance of the respiratory system after antigen challenge and eosinophils and lymphocytes in the OVA-S compared to the OVA group (p < 0.001, p = 0.003 and p < 0.001). Neither collagen nor elastic fiber contents were modified by stress in sensitized animals.. In this animal model, repeated stress amplified bronchoconstriction and inflammatory response in distal airways without interfering with extracellular matrix remodeling.

Topics: Administration, Inhalation; Adrenal Glands; Analysis of Variance; Animals; Disease Models, Animal; Extracellular Matrix; Guinea Pigs; Hypersensitivity; Inflammation; Male; Neutrophil Infiltration; Organ Size; Ovalbumin; Physical Stimulation; Respiration Disorders; Stress, Psychological; Swimming

Aiming the extract of Cordyceps sinensis significantly inhibits airway inflammation, airway hyperresponsiveness, and the infiltration of eosinophils in the airway of rats and may be related to the modulation of T helper (Th)1 and Th2 cells functions. The mechanisms of C. sinensis involved in modulation of suppression inflammation are not yet determined. In this study, the mechanism involved in the extract of C. sinensis-C.S.3-modulated suppression of inflammation was investigated in vivo and in vitro systems. The results showed that C.S.3 reduced airway inflammation in ovalbumin-induced allergic mice. Furthermore, we found C.S.3 could decrease extracellular signal-regulated kinase 1/2 signaling pathway to suppress activity of nuclear factor-κB in lung cells and cultured airway smooth muscle cells. Conclusion C.S.3 may provide clinical applications for asthma in the future.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cordyceps; Eosinophils; Extracellular Signal-Regulated MAP Kinases; Hypersensitivity; Inflammation; Male; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; NF-kappa B; Ovalbumin; Rats; Respiratory System

Pim kinases are a family of serine/threonine kinases whose activity can be induced by cytokines involved in allergy and asthma. These kinases play a role in cell survival and proliferation, but have not been examined, to the best of our knowledge, in the development of allergic disease. This study sought to determine the role of Pim1 kinase in the development of allergic airway responses. Mice were sensitized and challenged with antigen (primary challenge), or were sensitized, challenged, and rechallenged with allergen in a secondary model. To assess the role of Pim1 kinase, a small molecule inhibitor was administered orally after sensitization and during the challenge phase. Airway responsiveness to inhaled methacholine, airway and lung inflammation, cell composition, and cytokine concentrations were assessed. Lung Pim1 kinase concentrations were increased after ovalbumin sensitization and challenge. In the primary allergen challenge model, treatment with the Pim1 kinase inhibitor after sensitization and during airway challenges prevented the development of airway hyperresponsiveness, eosinophilic airway inflammation, and goblet cell metaplasia, and increased Th2 cytokine concentrations in bronchoalveolar fluid in a dose-dependent manner. These effects were also demonstrated after a secondary allergen challenge, where lung allergic disease was established before treatment. After treatment with the inhibitor, a significant reduction was evident in the number of CD4(+) and CD8(+) T cells and concentrations of cytokines in the airways. The inhibition of Pim1 kinase was effective in preventing the development of airway hyperresponsiveness, airway inflammation, and cytokine production in allergen-sensitized and allergen-challenged mice. These data identify the important role of Pim1 kinase in the full development of allergen-induced airway responses.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Enzyme Inhibitors; Female; Goblet Cells; Hypersensitivity; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Proto-Oncogene Proteins c-pim-1; Respiratory Hypersensitivity

Protease-activated receptor 2 (PAR-2) is a G protein-coupled receptor possibly involved in the pathogenesis of asthma. PAR-2 also modulates ion transport in cultured epithelial cells, but these effects in native airways are controversial. The influence of allergic inflammation on PAR-2-induced changes in ion transport has received little attention. Here, we studied immediate changes in transepithelial short circuit current (I (sc)) induced by PAR-2 activation in the tracheas of naive and allergic mice. Activation of PAR-2 with an apically added activation peptide (AP) induced a small increase in I (sc), while a much larger increase was observed following basolateral AP addition. In ovalbumin-sensitized and -challenged animals used as a model of allergic airway inflammation, the effect of basolateral AP addition was enhanced. Responses to basolateral AP in both naive and allergic mice were not decreased by blocking sodium absorption with amiloride or CFTR function with CFTR(inh)172 but were reduced by the cyclooxygenase inhibitor indomethacin and largely blocked (>80%) by niflumic acid, a calcium-activated chloride channels' (CaCC) blocker. Allergic mice also showed an enhanced response to ATP and thapsigargin. There was no change in mRNA expression of Par-2 or of the chloride channels Ano1 (Tmem16a) and Bestrophin 2 in tracheas from allergic mice, while mRNA levels of Bestrophin 1 were increased. In conclusion, basolateral PAR-2 activation in the mouse airways led to increased anion secretion through apical CaCC, which was more pronounced in allergic animals. This could be a protective mechanism aimed at clearing allergens and defending against mucus plugging.

Topics: Amiloride; Animals; Asthma; Benzoates; Bestrophins; Chloride Channels; Eye Proteins; Hypersensitivity; Indomethacin; Ion Channels; Male; Mice; Mice, Inbred BALB C; Niflumic Acid; Oligopeptides; Ovalbumin; Receptor, PAR-2; Thiazolidines; Tracheitis

MicroRNAs (miRNAs) are known to regulate the inflammatory response in various cell types. However, the ability of miRNAs to modulate dendritic cells (DCs) function for allergen immunotherapy is unclear.. To assess the role of miR-23b in the regulation of ovalbumin (OVA)-induced DC differentiation and function and to investigate the related molecular mechanisms.. Bone marrow-derived dendritic cells (BMDCs) were generated from murine bone marrow progenitor cells and subsequently stimulated with OVA to examine the profile of miRNA expression. After transfection with miR-23b reagents, DCs were evaluated for endocytic ability, surface marker expression, cytokine secretion and CD4+ T-cell differentiation. The possible roles of the Notch and NF-κB signalling pathways were also evaluated. Human monocyte-derived dendritic cells (MDDCs) were similarly evaluated as well.. Significant upregulation of miR-23b was observed in BMDCs pulsed with OVA. Following miR-23b transfection, BMDCs showed decreased OVA uptake, increased IL-10 production, decreased IL-12 production and an enhanced capacity to promote FoxP3+ CD4+ T regulatory cells (Tregs) differentiation. In addition, inactivation of the Notch1 and NF-κB signalling pathways were observed. Conversely, inhibition of miR-23b in BMDCs resulted in the opposite effects. In human MDDCs, miRNA23b transfection similarly increased IL-10 and decreased IL-12 production, and that treated human MDDCs induced increased FoxP3+ CD4+ T cells.. Our findings provide evidence that miR-23b is capable of inducing tolerogenic DC activity and Treg responses in vitro through the inhibition of the Notch1 and NF-κB signalling pathways; thus, miR-23b might represent a therapeutic target for the management of allergic diseases.

Topics: Allergens; Animals; Bone Marrow Cells; Cell Differentiation; Cells, Cultured; Dendritic Cells; Humans; Hypersensitivity; Immune Tolerance; Immunotherapy; Male; Mice; Mice, Inbred BALB C; MicroRNAs; NF-kappa B; Ovalbumin; Receptor, Notch1; Signal Transduction; Up-Regulation

The purpose of this study was to assess the effects of corticosteroid therapy on a murine model of allergic asthma using hyperpolarized (3)He magnetic resonance imaging (MRI) and respiratory mechanics measurements before, during, and after methacholine (MCh) challenge. Three groups of mice were prepared, consisting of ovalbumin sensitized/ovalbumin challenged (Ova/Ova, n = 5), Ova/Ova challenged but treated with the corticosteroid dexamethasone (Ova/Ova+Dex, n = 3), and ovalbumin-sensitized/saline-challenged (Ova/PBS, n = 4) control animals. All mice underwent baseline 3D (3)He MRI, then received a MCh challenge while 10 2D (3)He MR images were acquired for 2 min, followed by post-MCh 3D (3)He MRI. Identically treated groups underwent respiratory mechanics evaluation (n = 4/group) and inflammatory cell counts (n = 4/group). Ova/Ova animals exhibited predominantly large whole lobar defects at baseline, with significantly higher ventilation defect percentage (VDP = 19 ± 4%) than Ova/PBS (+2 ± 1%, P = 0.01) animals. Such baseline defects were suppressed by dexamethasone (0%, P = 0.009). In the Ova/Ova group, MCh challenge increased VDP on both 2D (+30 ± 8%) and 3D MRI scans (+14 ± 2%). MCh-induced VDP changes were diminished in Ova/Ova+Dex animals on both 2D (+21 ± 9%, P = 0.63) and 3D scans (+7 ± 2%, P = 0.11) and also in Ova/PBS animals on 2D (+6 ± 3%, P = 0.07) and 3D (+4 ± 1%, P = 0.01) scans. Because MCh challenge caused near complete cessation of ventilation in four of five Ova/Ova animals, even as large airways remained patent, this implies that small airway (<188 μm) obstruction predominates in this model. This corresponds with respiratory mechanics observations that MCh challenge significantly increases elastance and tissue damping but only modestly affects Newtonian airway resistance.

Topics: Adrenal Cortex Hormones; Airway Resistance; Animals; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Bronchoconstrictor Agents; Dexamethasone; Disease Models, Animal; Elasticity; Helium; Hypersensitivity; Isotopes; Lung; Magnetic Resonance Imaging; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Models, Biological; Ovalbumin; Pulmonary Ventilation; Respiratory Mechanics

Lactic acid bacteria (LAB) are microorganisms that benefit animals with allergic diseases and intestinal disorders such as inflammatory bowel disease. We propose that LAB can prevent cardiomyocytes inflammation and apoptosis in BALB/c mice using an ovalbumin (OVA)-induced allergy. Thirty-nine male BALB/c mice were divided into five groups: normal control, allergy control and three allergy groups each treated with Kefir I (Kefir I), Kefir II (Kefir II) or GM080 products (GM080). The myocardial architecture and apoptotic molecules in the excised left ventricle from these mice were investigated and post-treatment effects were evaluated. The inflammatory pathway, including toll-like receptor 4 (TLR4), phospholate-Jun-N-terminal kinase (p-JNK), JNK1/2 and tumor necrosis factor- alpha (TNF-α) and the mitochondria-dependent apoptosis phospholate-p38 (p-p38), Bcl-2 associated agonist of cell death (Bad), Bcl-2 associated X (Bax) and activated caspase 3, were found to be significant- ly increased in the hearts of allergy mice. The expression of phospholate-nuclear factor-κB (p-NFκB), TNF-α, p-p38 and Bad protein products were reduced or retarded in the Kefir I- or II-treated allergy group. The GM080-treated allergy group exhibited significantly lower p-JNK, JNK1/2, phospholate- Ikappa B (p-IκB), Bax and Bad protein products than the Kefir I and Kefir II allergy groups. These results indicate that LAB can reduce inflammation and prevent apoptosis of cardiomyocytes in the heart of OVA-induced allergy mice.

Topics: Animals; Apoptosis; Caspase 3; Cytochromes c; Heart Ventricles; Hypersensitivity; Lactobacillus; Male; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Mitochondria, Heart; Myocarditis; Myocardium; Ovalbumin; Probiotics; Proto-Oncogene Proteins c-bcl-2; Toll-Like Receptor 4

The anti-inflammatory effects of low-molecular weight chitosan oligosaccharides (LM-COS) prepared from high-molecular weight chitosan by enzymatic digestion were investigated against allergic reaction and allergic asthma in vivo and in vitro. Allergic asthma is an inflammatory disease of the airways associated with enhanced degranulation and cytokine generation. The LM-COS (<1 kDa), consisting of glucosamine (GlcN)(n), n=3-5, were capable of inhibiting both antigen-stimulated degranulation and cytokine generation in rat basophilic leukemia RBL-2H3 cells. The protective effect of LM-COS against ovalbumin (OVA)-induced lung inflammation in asthma model mice was also examined. Oral administration of LM-COS (16 mg/kg body weight/day) resulted in a significant reduction in both mRNA and protein levels of interleukin (IL)-4, IL-5, IL-13, tumor necrosis factor (TNF)-α in the lung tissue and bronchoalveolar lavage fluid (BALF); The protein levels of IL-4, IL-13 and TNF-α in BALF were decreased by 5.8-fold, 3.0-fold and 9.9-fold, respectively, compared to those in the OVA-sensitized/challenged asthma control group. These results suggest that the oral administration of LM-COS is effective in alleviating the allergic inflammation in vivo and thus can be a good source material for the development of a potent therapeutic agent against mast cell-mediated allergic inflammatory responses and airway inflammation in allergic inflammatory diseases, including asthma.

Topics: Animals; Anti-Inflammatory Agents; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Cell Line, Tumor; Chitosan; Disease Models, Animal; Female; Glucosamine; Hypersensitivity; Immunoglobulin E; Interleukins; Leukemia, Basophilic, Acute; Lung; Mice; Mice, Inbred BALB C; Molecular Weight; Oligosaccharides; Ovalbumin; Tumor Necrosis Factor-alpha

The IL-4 receptor α (IL-4Rα) chain has a broad expression pattern and participates in IL-4 and IL-13 signaling, allowing it to influence several pathological components of allergic lung inflammation. We previously reported that IL-4Rα expression on both bone marrow-derived and non-bone marrow-derived cells contributed to the severity of allergic lung inflammation. There was a correlation between the number of macrophages expressing the IL-4Rα, CD11b, and IA(d), and the degree of eosinophilia in ovalbumin challenged mice. The engagement of the IL-4Rα by IL-4 or IL-13 is able to stimulate the alternative activation of macrophages (AAM). The presence of AAM has been correlated with inflammatory responses to parasites and allergens. Therefore, we hypothesized that IL-4Rα⁺ AAM play an active role in allergic lung inflammation. To directly determine the role of AAM in allergic lung inflammation, M-CSF-dependent macrophages (BMM) were prepared from the bone-marrow of IL-4Rα positive and negative mice and transferred to IL-4RαxRAG2(-/-) mice. Wild type TH2 cells were provided exogenously.. Mice receiving IL-4Rα(+/+) BMM showed a marked increase in the recruitment of eosinophils to the lung after challenge with ovalbumin as compared to mice receiving IL-4Rα(-/-) BMM. As expected, the eosinophilic inflammation was dependent on the presence of TH2 cells. Furthermore, we observed an increase in cells expressing F4/80 and Mac3, and the AAM marker YM1/2 in the lungs of mice receiving IL-4Rα(+/+) BMM. The BAL fluid from these mice contained elevated levels of eotaxin-1, RANTES, and CCL2.. These results demonstrate that transfer of IL-4Rα + macrophages is sufficient to enhance TH2-driven, allergic inflammation. They further show that stimulation of macrophages through IL-4Rα leads to their alternative activation and positive contribution to the TH2-driven allergic inflammatory response in the lung. Since an increase in AAM and their products has been observed in patients with asthma exacerbations, these results suggest that AAM may be targeted to alleviate exacerbations.

Topics: Adoptive Transfer; Animals; Bone Marrow Transplantation; Bronchoalveolar Lavage Fluid; Chickens; Disease Models, Animal; Eosinophils; Humans; Hypersensitivity; Interleukin-4 Receptor alpha Subunit; Lung; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Phenotype; Pneumonia; Staining and Labeling; Th2 Cells; Tumor Necrosis Factor-alpha

Nilotinib is a new orally bioavailable potent tyrosine kinase inhibitor that is used for the treatment of BCR-ABL-positive chronic myelogenous leukemia. However, its effect on mast cell-mediated anaphylactic reaction is still not known. The present study aimed to investigate the effect of nilotinib on the anaphylactic allergic reaction and study its possible mechanism(s) of action. Nilotinib administration prevented systemic anaphylaxis in mice, mediated by compound 48/80, in a dose- and time-dependent manner. Also, nilotinib significantly inhibited (P<0.05) allergic paw edema in rats. Furthermore, nilotinib significantly decreased (P<0.05) the IgE-mediated passive cutaneous anaphylaxis in a dose dependent manner. In addition, nilotinib dose-dependently reduced histamine release from the rat peritoneal mast cells activated either by compound 48/80 or by ovalbumin. Moreover, nilotinib attenuated the secretion of pro-inflammatory cytokine, tumor necrosis factor (TNF)-α expression in the rat peritoneal mast cells. These findings provide evidence that nilotinib inhibits mast cell-derived immediate-type allergic reactions and so it could be a candidate as an anti-allergic agent.

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Cytokines; Edema; Histamine Release; Hypersensitivity; Immunoglobulin E; Male; Mast Cells; Mice; Ovalbumin; p-Methoxy-N-methylphenethylamine; Protein-Tyrosine Kinases; Pyrimidines; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

Previous studies in young rats reported the impact of cocoa intake on healthy immune status and allow suggesting it may have a role in the prevention of some immune-mediated diseases. The aim of this study was to ascertain the effect of a cocoa diet in a model of allergy in young rats. Three-week-old Brown Norway rats were immunized by i.p. injection of ovalbumin (OVA) with alum as adjuvant and Bordetella pertussis toxin. During the next 4 weeks rats received either a cocoa diet (containing 0.2% polyphenols, w/w) or a standard diet. Animals fed a standard diet showed high concentrations of anti-OVA IgG1, IgG2a, IgG2b and high anti-OVA IgE titres, which is the antibody involved in allergic response. In contrast, animals fed a cocoa diet showed significantly lower concentrations of anti-OVA IgG1 and IgG2a antibodies. Interestingly, the cocoa diet prevented anti-OVA IgE synthesis and decreased total serum IgE concentration. Analysis of cytokine production in lymph node cells at the end of the study revealed that, in this compartment, the cocoa diet decreased the tumor necrosis factor (TNF)-α and the interleukin (IL)-10 secretion but not IL-4 production. In conclusion, a cocoa-enriched diet in young rats produces an immunomodulatory effect that prevents anti-allergen IgE synthesis, suggesting a potential role for cocoa flavonoids in the prevention or treatment of allergic diseases.

Topics: Alum Compounds; Animals; Anti-Allergic Agents; Body Weight; Cacao; Diet; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Interleukin-10; Interleukin-4; Lymph Nodes; Ovalbumin; Pertussis Toxin; Polyphenols; Rats; Rats, Inbred BN; Time Factors; Tumor Necrosis Factor-alpha

Despite the existing knowledge regarding the neuropathology of Alzheimer's disease (AD), the cause of sporadic forms of the disease is unknown. It has been suggested that systemic inflammation may have a role, but the exact mechanisms through which inflammatory processes influence the pathogenesis and progress of AD are not obvious. Allergy is a chronic inflammatory disease affecting more than 20% of the Western population, but the effects of allergic conditions on brain functions are largely unknown. The aim of this study was to investigate whether or not chronic peripheral inflammation associated with allergy affects the expression of AD-related proteins and inflammatory markers in the brain. On the basis of previously described models for allergy in mice we developed a model of chronic airway allergy in mouse, with ovalbumin as allergen. The validity of the chronic allergy model was confirmed by a consistent and reproducible eosinophilia in the bronchoalveolar lavage (BAL) fluid of allergic animals. Allergic mice were shown to have increased brain levels of both immunoglobulin (Ig) G and IgE with a widespread distribution. Allergy was also found to increase phosphorylation of tau protein in the brain. The present data support the notion that allergy-dependent chronic peripheral inflammation modifies the brain inflammatory status, and influences phosphorylation of an AD-related protein, indicating that allergy may be yet another factor to be considered for the development and/or progression of neurodegenerative diseases such as AD.

Topics: Allergens; Alzheimer Disease; Animals; Brain; Bronchoalveolar Lavage Fluid; Chronic Disease; Disease Models, Animal; Eosinophilia; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; tau Proteins

Since the Gβγ subunit of Gi protein has been importantly implicated in regulating immune and inflammatory responses, this study investigated the potential role and mechanism of action of Gβγ signaling in regulating the induction of airway hyperresponsiveness (AHR) in a rabbit model of allergic asthma. Relative to non-sensitized animals, OVA-sensitized rabbits challenged with inhaled OVA exhibited AHR, lung inflammation, elevated BAL levels of IL-13, and increased airway phosphodiesterase-4 (PDE4) activity. These proasthmatic responses were suppressed by pretreatment with an inhaled membrane-permeable anti-Gβγ blocking peptide, similar to the suppressive effect of glucocorticoid pretreatment. Extended mechanistic studies demonstrated that: 1) corresponding proasthmatic changes in contractility exhibited in isolated airway smooth muscle (ASM) sensitized with serum from OVA-sensitized+challenged rabbits or IL-13 were also Gβγ-dependent and mediated by MAPK-upregulated PDE4 activity; and 2) the latter was attributed to Gβγ-induced direct stimulation of the non-receptor tyrosine kinase, c-Src, resulting in downstream activation of ERK1/2 and its consequent transcriptional upregulation of PDE4. Collectively, these data are the first to identify that a mechanism involving Gβγ-induced direct activation of c-Src, leading to ERK1/2-mediated upregulation of PDE4 activity, plays a decisive role in regulating the induction of AHR and inflammation in a rabbit model of allergic airway disease.

Topics: Animals; Asthma; Cyclic Nucleotide Phosphodiesterases, Type 4; GTP-Binding Protein beta Subunits; GTP-Binding Protein gamma Subunits; Humans; Hypersensitivity; Hypersensitivity, Immediate; Inflammation; Male; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Models, Biological; Muscle, Smooth; Ovalbumin; Rabbits; Respiratory System; Signal Transduction; Transcription, Genetic

To investigate the effect of Asian sand dust (ASD) on mucin production in human respiratory epithelial cells in vitro and in allergic murine nasal epithelial cells.. Controlled, in vitro.. Academic research laboratory.. Human NCI-H292 cells were treated with ASD and analyzed by immunostaining, reverse transcriptase-polymerase chain reaction for MUC5AC mRNA expression, and Periodic Acid Schiff (PAS) staining. Forty female BALB/c mice were classified into 4 groups. Two groups were sensitized with ovalbumin (OVA), and 1 of these was treated with ASD (ASD+OVA). The 2 nonsensitized groups were treated with ASD or saline. Then the murine nasal mucosal tissues were evaluated by hematoxylin and eosin (H&E) staining, PAS staining, and immunostaining for MUC5AC and transforming growth factor (TGF)-α proteins.. The numbers of MUC5AC-immunopositive NCI-H292 cells and PAS-positive NCI-H292 cells were significantly higher in the ASD-treated cells than in the control cells (P = .039 and P = .029, respectively). MUC5AC mRNA expression in the cells increased with increasing concentrations of ASD. In the murine nasal epithelial tissues, the numbers of eosinophils and PAS-positive cells were significantly higher in the ASD+OVA group than in the OVA group (H&E staining, P = .037; PAS staining, P = .019). At 2 weeks, the numbers of MUC5AC- and TGF-α-positive cells in the nasal epithelial tissue were significantly higher in the ASD+OVA group than in the OVA group (P = .031 and P = .033, respectively).. ASD can induce mucin production in respiratory epithelial cells.

Topics: Animals; Cell Culture Techniques; Disease Models, Animal; Dust; Epithelial Cells; Female; Humans; Hypersensitivity; Mice; Mice, Inbred BALB C; Mucin 5AC; Ovalbumin; Respiratory Mucosa; Silicon Dioxide; Transforming Growth Factor alpha

Elevated PGE(2) is a hallmark of most inflammatory lesions. This lipid mediator can induce the cardinal signs of inflammation, and the beneficial actions of nonsteroidal anti-inflammatory drugs are attributed to inhibition of cyclooxygenase (COX)-1 and COX-2, enzymes essential in the biosynthesis of PGE(2) from arachidonic acid. However, both clinical studies and rodent models suggest that, in the asthmatic lung, PGE(2) acts to restrain the immune response and limit physiological change secondary to inflammation. To directly address the role of PGE(2) in the lung, we examined the development of disease in mice lacking microsomal PGE(2) synthase-1 (mPGES1), which converts COX-1/COX-2-derived PGH(2) to PGE(2). We show that mPGES1 determines PGE(2) levels in the naive lung and is required for increases in PGE(2) after OVA-induced allergy. Although loss of either COX-1 or COX-2 increases the disease severity, surprisingly, mPGES1(-/-) mice show reduced inflammation. However, an increase in serum IgE is still observed in the mPGES1(-/-) mice, suggesting that loss of PGE(2) does not impair induction of a Th2 response. Furthermore, mPGES1(-/-) mice expressing a transgenic OVA-specific TCR are also protected, indicating that PGE(2) acts primarily after challenge with inhaled Ag. PGE(2) produced by the lung plays the critical role in this response, as loss of lung mPGES1 is sufficient to protect against disease. Together, this supports a model in which mPGES1-dependent PGE(2) produced by populations of cells native to the lung contributes to the effector phase of some allergic responses.

Topics: Animals; Cell Proliferation; Cyclooxygenase 1; Cyclooxygenase 2; Cytokines; Dinoprostone; Female; Hypersensitivity; Inflammation; Intramolecular Oxidoreductases; Lung; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Prostaglandin-E Synthases; Receptors, Antigen, T-Cell; Th2 Cells

Epidemiological studies suggest that viral infections during childhood are a risk factor for the development of asthma. However, the role of virus-specific pattern recognition receptors in this process is not well defined. In the current study, we compare the effects of the inhaled viral TLR ligands polyinosinic-polycytidylic acid (TLR3) and resiquimod (TLR7/8) on sensitization to a model allergen (OVA) in a murine model. Both compounds enhance the migration, activation, and Ag-processing of myeloid dendritic cells from the lung to the draining lymph nodes comparable to the effects of LPS. Application of polyinosinic-polycytidylic acid [poly(I:C)] or LPS induces production of allergen-specific IgE and IgG1, whereas resiquimod (R848) had no effect. In addition, rechallenge of mice with OVA resulted in airway inflammation and mucus production in animals that received either poly(I:C) or LPS but not after application of R848. In summary, these results show that activation of TLR3 in combination with inhaled allergen results in induction of dendritic cell activation and migration similar to the effects of LPS. This leads to the development of allergic airway disease after allergen rechallenge, whereas mice treated with R848 did not develop allergic airway disease. These findings give further insight into the effects of stimulation of different TLRs on the development of asthma.

Topics: Administration, Inhalation; Allergens; Animals; Dendritic Cells; Disease Models, Animal; Hypersensitivity; Imidazoles; Ligands; Lipopolysaccharides; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Ovalbumin; Poly I-C; Toll-Like Receptor 3; Toll-Like Receptor 7; Toll-Like Receptor 8

Under inflammatory conditions, T cell-dependent (TD) protein antigens induce proinflammatory T- and B-cell responses. In contrast, tolerance induction by TD antigens without costimulation triggers the development of regulatory T cells. Under both conditions, IgG antibodies are generated, but whether they have different immunoregulatory functions remains elusive.. It was shown recently that proinflammatory or anti-inflammatory effector functions of IgG molecules are determined by different Fc N-linked glycosylation patterns. We sought to examine the Fc glycosylation and anti-inflammatory quality of IgG molecules formed on TD tolerance induction.. We administered chicken ovalbumin (OVA) with or without costimulus to mice and analyzed OVA-reactive IgG Fc glycosylation. The anti-inflammatory function of differentially glycosylated anti-OVA IgGs was further investigated in studies with dendritic cell cultures and in an in vivo model of allergic airway disease. Additionally, we analyzed the Fc glycosylation pattern of birch pollen-reactive serum IgGs after successful allergen-specific immunotherapy in patients.. Stimulation with TD antigens under inflammatory conditions induces plasma cells expressing low levels of α2,6-sialyltransferase and producing desialylated IgGs. In contrast, plasma cells induced on tolerance induction did not downregulate α2,6-sialyltransferase expression and secreted immunosuppressive sialylated IgGs that were sufficient to block antigen-specific T- and B-cell responses, dendritic cell maturation, and allergic airway inflammation. Importantly, successful specific immunotherapy in allergic patients also induced sialylated allergen-specific IgGs.. Our data show a novel antigen-specific immunoregulatory mechanism mediated by anti-inflammatory sialylated IgGs that are formed on TD tolerance induction. These findings might help to develop novel antigen-specific therapies for the treatment of allergy and autoimmunity.

Topics: Animals; Antigen-Antibody Complex; Antigens; beta-D-Galactoside alpha 2-6-Sialyltransferase; Desensitization, Immunologic; Epitopes; Female; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin G; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Plasma Cells; Receptors, IgG; Sialyltransferases; T-Lymphocytes

The Th1/Th2 balance represents an important factor in the pathogenesis of renal ischemia-reperfusion injury (IRI). In addition, IRI causes a systemic inflammation that can affect other tissues, such as the lungs. To investigate the ability of renal IRI to modulate pulmonary function in a specific model of allergic inflammation, C57Bl/6 mice were immunized with ovalbumin/albumen on days 0 and 7 and challenged with an ovalbumin (OA) aerosol on days 14 and 21. After 24 h of the second antigen challenge, the animals were subjected to 45 minutes of ischemia. After 24 h of reperfusion, the bronchoalveolar lavage (BAL) fluid, blood and lung tissue were collected for analysis. Serum creatinine levels increased in both allergic and non-immunized animals subjected to IRI. However, BAL analysis showed a reduction in the total cells (46%) and neutrophils (58%) compared with control allergic animals not submitted to IRI. In addition, OA challenge induced the phosphorylation of ERK and Akt and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lung homogenates. After renal IRI, the phosphorylation of ERK and expression of COX-2 and iNOS were markedly reduced; however, there was no difference in the phosphorylation of Akt between sham and ischemic OA-challenged animals. Mucus production was also reduced in allergic mice after renal IRI. IL-4, IL-5 and IL-13 were markedly down-regulated in immunized/challenged mice subjected to IRI. These results suggest that renal IRI can modulate lung allergic inflammation, probably by altering the Th1/Th2 balance and, at least in part, by changing cellular signal transduction factors.

Topics: Animals; Blood Cell Count; Bronchoalveolar Lavage Fluid; Creatinine; Cyclooxygenase 2; Hypersensitivity; Inflammation; Interleukins; Kidney; Lung; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Mucus; Neutrophils; Nitric Oxide Synthase Type II; Ovalbumin; Phosphorylation; Reperfusion Injury; Th1-Th2 Balance

Eosinophilic inflammation is a hallmark of chronic rhinosinusitis with nasal polyps. To model this disease process experimentally, nasal sensitization of mice with ovalbumin or aspergillus has been described. Here, we describe a genetically mutant mouse that develops robust spontaneous nasal eosinophilic inflammation. These mice lack the enzyme SHP-1 that down-regulates the IL-4Rα/stat6 signaling pathway. We compared nasal inflammation and inflammatory mediators in SHP-1 deficient mice (mev) and an ovalbumin-induced nasal allergy model.. A novel technique of trans-pharyngeal nasal lavage was developed to obtain samples of inflammatory cells from the nasal passages of allergic and mev mice. Total and differential cell counts were performed on cytospin preparations. Expression of tissue mRNA for IL-4, IL-13, and mouse beta-defensin-1 (MBD-1) was determined by quantitative PCR. Eotaxin in the lavage fluid was assessed by ELISA.. Allergic and mev mice had increased total cells and eosinophils compared with controls. Expression of IL-4 was similarly increased in both allergic and mev mice, but expression of IL-13 and eotaxin was significantly greater in the allergic mice than mev mice. Eotaxin was significantly up-regulated in both allergic rhinitis and mev mice. In both models of eosinophilic inflammation, down-regulation of the innate immune marker MBD-1 was observed.. The mev mice display spontaneous chronic nasal eosinophilic inflammation with potential utility for chronic rhinosinusitis with nasal polyps research. The eosinophilic infiltrate is more robust in the mev mice than allergic mice, but Th2 cytokine expression is not as pronounced. Decreased MBD-1 expression in both models supports the concept that Th2-cytokines down-regulate sinonasal innate immunity in humans, and suggests a role for mouse models in investigating the interaction between adaptive and innate immunity in the sinonasal mucosa.

Topics: Animals; beta-Defensins; Eosinophils; Gene Expression Regulation; Hypersensitivity; Inflammation; Interleukin-13; Interleukin-4; Mice; Mutation; Nasal Lavage Fluid; Nasal Mucosa; Nose Diseases; Ovalbumin; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Signal Transduction; STAT6 Transcription Factor

Aerobic conditioning (AC) performed either during or after sensitization reduces allergic inflammation in mice; however, the effects of AC performed before and during allergic sensitization on airway inflammation are unknown. Mice were divided into Control, AC, OVA, and AC + OVA groups. Mice were trained in a treadmill followed by either ovalbumin (OVA) sensitization or saline administration. Peribronchial inflammation, OVA-specific IgE and IgG1 titers, the expression of Th1 and Th2 cytokines, and airway remodeling were evaluated, as well as the expression of Eotaxin, RANTES, ICAM-1, VCAM-1, TGF-β and VEGF. Aerobic conditioning performed before and during allergic sensitization displayed an inhibitory effect on the OVA-induced migration of eosinophils and lymphocytes to the airways, a reduction of IgE and IgG1 titers and an inhibition of the expression of Th2 cytokines. The AC + OVA group also demonstrated reduced expression of ICAM-1, VCAM-1, RANTES, TGF-β and VEGF, as well as decreased airway remodeling (p<0.05). The effects of AC before and during the sensitization process inhibit allergic airway inflammation and reduce the production of Th2 cytokines and allergen-specific IgE and IgG1.

Topics: Animals; Cytokines; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Physical Conditioning, Animal; Pneumonia; Sodium Chloride

The aim of this study is to investigate the reciprocal effect of Staphylococcus aureus colonization and allergic rhinitis in an allergy model of mice.. BALB/c mice with intraperitoneal ovalbumin (OVA) sensitization and/or intranasal S. aureus inoculation were prepared. The following 4 groups were designed: an OVA-sensitized S. aureus-inoculated (AR-SA) group, an OVA-sensitized uninoculated (AR) group, a nonsensitized S. aureus-inoculated (SA) group, and a nonsensitized uninoculated (control) group. After intranasal OVA challenge, nasal lavage fluid, peripheral blood, and nasal mucosa were collected. Polymorphonuclear cells in the nasal lavage fluid were counted, serum OVA-specific IgE and IgG1 were measured by enzyme immunoassays, and IL-4, IL-5, and IFN-γ mRNAs in the nasal mucosa were assessed by quantitative real-time reverse transcription-PCR. The number of S. aureus in the nasal mucosa and lavage fluid was counted.. Both eosinophil and neutrophil counts were larger in the AR-SA group than in the other groups. Both IgE and IgG1 levels were higher in the AR and AR-SA groups than in the SA and control groups, and the IgG1 level was higher in the AR-SA group than in the AR group. The expression of IL-4 mRNA was higher in the AR-SA group than in the other groups, and the expression of IL-5 mRNA was higher in the AR-SA group than in the SA group. The AR-SA group showed higher counts of S. aureus in the nasal mucosa than the SA group.. These results indicate the mutually potentiating effect of S. aureus colonization and allergic rhinitis.

Topics: Allergens; Animals; Bacterial Load; Cytokines; Eosinophils; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Nasal Cavity; Nasal Lavage Fluid; Nasal Mucosa; Neutrophils; Ovalbumin; Staphylococcal Infections; Staphylococcus aureus

Inflammatory mediators from peripheral tissues may control dendritic cell (DC) development in the bone marrow. In this study, DCs (CD11c(+) cells) differentiated from the bone marrow of mice with inflammation of the airways, or the peritoneal cavity had poor priming ability resulting in reduced, long-lived responses to that antigen in vivo. This indicates enhancement of regulatory mechanisms of immune responses through a peripheral tissue-bone marrow axis. If CD11c(+) cells, expanded from the bone marrow of mice with tissue inflammation were antigen pre-loaded and injected into mice already sensitized to that antigen, then subsequent contact hypersensitivity responses were significantly reduced. The effects of inflammation were imprinted in vivo and were independent of in vitro culture conditions for DC differentiation. The effect of tissue inflammation on the bone marrow DC precursors was not detected in mice treated subcutaneously with slow-release indomethacin pellets, suggesting a role for prostanoids, including prostaglandin E(2), in differentiation of regulatory CD11c(+) cells from bone marrow. Our study represents an important homeostatic process with potential for therapeutic use in the future.

Topics: Alum Compounds; Animals; B7-2 Antigen; Bone Marrow Cells; CD11c Antigen; Cell Differentiation; Cell Movement; Cross-Priming; Dendritic Cells; Granulocyte-Macrophage Colony-Stimulating Factor; Haptens; Homeostasis; Hypersensitivity; Immunization; Indomethacin; Inflammation; Interleukin-4; Lung; Lung Diseases; Lymph Nodes; Membrane Proteins; Mice; Mice, Inbred BALB C; Myelopoiesis; Ovalbumin; Peritoneal Cavity

Intravenous immunoglobulin (IVIG) displays anti-inflammatory activities in many diseases. Subcutaneous administration of anti-IgE in humans provides benefit in severe persistent allergic asthma. Given the well established efficacy of sublingual allergen immunotherapy in respiratory type I allergies, we investigated the therapeutic potential of sublingual immunoglobulin (SLIG), most particularly anti-IgE SLIG, in a murine model of allergen-driven airway inflammation.. BALB/c mice sensitized with ovalbumin (OVA) were treated sublingually with rat monoclonal IgG1 or IgG2a, either directed to mouse IgE or with no reported specificity. Airway hyperresponsiveness (AHR) was assessed by whole body plethysmography, and eosinophil infiltrates were characterized in bronchial alveolar lavages (BAL). OVA-specific antibody and T cell responses were analyzed in sera and saliva or lung and draining lymph nodes, by ELISA or CBA measurement of cytokine production, respectively.. AHR and BAL eosinophil infiltrates were substantially decreased in mice treated sublingually with particulate OVA (positive control), as well as in animals receiving various rat IgG1, irrespective of their specificity for murine IgE. In contrast, no improvement was observed in mice treated with PBS (negative control) or various rat IgG2a. SLIG anti-inflammatory activity is not related to a downregulation of Th2, Th17 or an induction of Foxp3(+) CD4(+) regulatory T cell responses. Mass spectrometry analysis of glycan moieties, such as sialic acid, suggests that the differential efficacy of rat IgG1 and IgG2a is not related to their capacity to interact with lectins borne by oral immune cells.. In a murine model of allergen-driven airway inflammation, SLIG exhibits an anti-inflammatory activity irrespective of the immunoglobulin specificity, and in the absence of allergen. As a noninvasive approach, SLIG deserves to be further studied as a treatment for other inflammatory diseases beyond allergic asthma.

Topics: Administration, Sublingual; Animals; Anti-Inflammatory Agents; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Humans; Hypersensitivity; Immunoglobulin G; Immunotherapy; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plethysmography, Whole Body; Respiratory Tract Diseases; Treatment Outcome

Allergy is an inflammation associated with an elevated T helper (Th) 2 lymphocyte responses to allergens and elevated serum IgE levels and cytokines. In one of our previous studies using a cell model, various flavonoids were found to be involved the anti-inflammatory effects of adlay bran. The present study investigated the effect of the ethyl-acetate fraction of ethanolic extract of adlay bran (ABE-EtOAc) in an ovalbumin (OVA)-immunized murine model. Six-week-old female BALB/c mice underwent OVA sensitization and were used as an allergy model. An orogastric gavage was used to force feed these mice with 240 mg/kg ABE-EtOAc from their sixth week through their twelfth week. Immune reactions were determined by measuring changes in Th2-type cytokine (IL-4 and IL-5) levels and production of antibodies. ABE-EtOAc was found capable of regulating the Th1/Th2 immune reaction through its regulation of IL-2 and IL-4. It also significantly reduced the production of anti-OVA IgE antibodies (10%), increased the secretion of IFN-γ and decreased the secretion of IL-6 (38%). These results suggest that adlay bran extract can reduce an allergic reaction by balancing Th1/Th2 immune responses and that it might be used in the treatment of this condition.

Topics: Animal Feed; Animals; Coix; Cytokines; Ethanol; Female; Gene Expression Regulation; Hypersensitivity; Immunity, Innate; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Seeds; Spleen

Serine proteases promote inflammation and tissue remodeling by activating proteinase-activated receptors, urokinase, metalloproteinases and angiotensin. In the present study, 4-(2-Aminoethyl) benzenesulfonyl fluoride (AEBSF) a serine protease inhibitor was evaluated for prophylactic and therapeutic treatment in mouse model of airway allergy.. BALB/c mice were sensitized by i.p route and challenged with ovalbumin. They were treated i.n. with 2, 10 and 50 µg of AEBSF, one hour before or after challenge and euthanized to collect BALF (bronchoalveolar lavage fluid), blood and lungs. Proteolytic activity, total cell/eosinophil/neutrophil count eosinophil peroxidase activity (EPO), IL-4, IL-5, IL-10, IL-13, cysteinyl leukotrienes and 8-isoprostane were determined in BALF and immunoglobulins were measured in serum. H&E and PAS stained lung sections were examined for cellular infiltration and airway inflammation.. Mice exposed to ovalbumin and treated with PBS showed increased cellular infiltration in lungs and higher serum IgE, IgG1 and IgG2a levels as compared to sham mice. Treatment with AEBSF reduced total cells/eosinophil/neutrophil infiltration. Both prophylactic and therapeutic AEBSF treatment of 10 or 50 µg reduced serum IgE and IgG1 significantly (p<0.05) than control. AEBSF treatment reduced the proteolytic activity in BALF. IL-4 IL-5 and IL-13 levels decreased significantly (p<0.05) after AEBSF treatment while IL-10 levels increased significantly (p<0.05) in BALF. Airway inflammation and goblet cell hyperplasia reduced as demonstrated by lung histopathology, EPO activity and cysteinyl leukotrienes in BALF after treatment. AEBSF treatment also suppressed oxidative stress in terms of 8-isoprostane in BALF. Among the treatment doses, 10 or 50 µg of AEBSF were most effective in reducing the inflammatory parameters.. Prophylactic and therapeutic treatment with serine protease inhibitor attenuates the airway inflammation in mouse model of airway allergy and have potential for adjunct therapy.

Topics: Animals; Bronchoalveolar Lavage Fluid; Eosinophil Peroxidase; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-10; Interleukin-4; Interleukin-5; Leukotrienes; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Oxidative Stress; Serine Proteinase Inhibitors; Sulfones

IL-4 produced by Th2 cells can block cytokine production by Th1 cells, and Th1 IFN-γ is known to counterregulate Th2 immune response, inhibiting allergic eosinophilia. As intrauterine undernutrition can attenuate lung inflammation, we investigated the influence of intrauterine undernourishment on the Th1/Th2 cytokine balance and allergic lung inflammation. Intrauterine undernourished offspring were obtained from dams fed 50% of the nourished diet of their counterparts and were immunized at 9 weeks of age. We evaluated the cell counts and cytokine protein expression in the bronchoalveolar lavage, mucus production and collagen deposition, and cytokine gene expression and transcription factors in lung tissue 21 days after ovalbumin immunization. Intrauterine undernourishment significantly reduced inflammatory cell airway infiltration, mucus secretion and collagen deposition, in rats immunized and challenged. Intrauterine undernourished rats also exhibited an altered cytokine expression profile, including higher TNF-α and IL-1β expression and lower IL-6 expression than well-nourished rats following immunization and challenge. Furthermore, the intrauterine undernourished group showed reduced ratios of the IL-4/IFN-γ and the transcription factors GATA-3/T-Bet after immunization and challenge. We suggest that the attenuated allergic lung inflammation observed in intrauterine undernourished rats is related to an altered Th1/Th2 cytokine balance resulting from a reduced GATA-3/T-bet ratio.

Topics: Animals; Bronchoalveolar Lavage Fluid; Female; GATA3 Transcription Factor; Hypersensitivity; Interferon-gamma; Interleukin-1beta; Interleukin-4; Interleukin-6; Male; Malnutrition; Ovalbumin; Pneumonia; Pregnancy; Prenatal Exposure Delayed Effects; Prenatal Nutritional Physiological Phenomena; Rats; Rats, Wistar; T-Box Domain Proteins; Th1 Cells; Th1-Th2 Balance; Th2 Cells; Tumor Necrosis Factor-alpha

Clinical reports indicate that patients with allergy/asthma commonly have associated symptoms of anxiety/depression. Anxiety/depression can be reduced by 5-hydroxytryptophan (5-HTP) supplementation. However, it is not known whether 5-HTP reduces allergic inflammation. Therefore, we determined whether 5-HTP supplementation reduces allergic inflammation. We also determined whether 5-HTP decreases passage of leukocytes through the endothelial barrier by regulating endothelial cell function. For these studies, C57BL/6 mice were supplemented with 5-HTP, treated with ovalbumin fraction V (OVA), house dust mite (HDM) extract, or IL-4, and examined for allergic lung inflammation and OVA-induced airway responsiveness. To determine whether 5-HTP reduces leukocyte or eosinophil transendothelial migration, endothelial cells were pretreated with 5-HTP, washed and then used in an in vitro transendothelial migration assay under laminar flow. Interestingly, 5-HTP reduced allergic lung inflammation by 70-90% and reduced antigen-induced airway responsiveness without affecting body weight, blood eosinophils, cytokines, or chemokines. 5-HTP reduced allergen-induced transglutaminase 2 (TG2) expression and serotonylation (serotonin conjugation to proteins) in lung endothelial cells. Consistent with the regulation of endothelial serotonylation in vivo, in vitro pretreatment of endothelial cells with 5-HTP reduced TNF-α-induced endothelial cell serotonylation and reduced leukocyte transendothelial migration. Furthermore, eosinophil and leukocyte transendothelial migration was reduced by inhibitors of transglutaminase and by inhibition of endothelial cell serotonin synthesis, suggesting that endothelial cell serotonylation is key for leukocyte transendothelial migration. In summary, 5-HTP supplementation inhibits endothelial serotonylation, leukocyte recruitment, and allergic inflammation. These data identify novel potential targets for intervention in allergy/asthma.

Topics: 5-Hydroxytryptophan; Animals; Antidepressive Agents, Second-Generation; Asthma; Cell Adhesion Molecules; Cell Line; Cell Movement; Chemokines; Disease Models, Animal; Endothelial Cells; Female; Hypersensitivity; Immunosuppression Therapy; Interleukin-4; Leukocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Pyroglyphidae; Serotonin; Spleen

Goblet cell hyperplasia (GCH) and mucous hypersecretion are common pathological features of chronic pulmonary diseases, including asthma, chronic obstructive pulmonary disease (COPD), lung cancer, and cystic fibrosis. Despite numerous studies, the molecular basis for this condition remains elusive. Gab2 is a member of the Dos/Gab subfamily scaffolding molecules and plays important roles in regulating growth, differentiation, and inflammation. We found that an elevated level of Gab2 correlates with up-regulated mucus in airway epithelia from patients with lung cancer or COPD, suggesting the potential involvement of Gab2 in pathological lesions in lungs. Knockdown of Gab2 in human airway epithelial cells in vitro decreases IL-13-induced expression of mucin genes. To address the in vivo role of Gab2 in lungs, Gab2-knockout (Gab2(-/-)) mice were sensitized and challenged with ovalbumin (OVA). Further analysis of lungs in an OVA-induced allergy model suggested that GCH and mucus production are remarkably reduced in Gab2(-/-) mice. Mechanistically, Gab2 positively regulates IL-13-induced activation of TYK2/STAT6 by decreasing SOCS3-mediated degradation of TYK2. Together, we define a novel role for Gab2 in mediating mucin gene expression and GCH; these findings have important implications for the pathogenesis and therapy of airway inflammatory diseases.

Topics: Adaptor Proteins, Signal Transducing; Animals; Epithelial Cells; Gene Expression Regulation; Goblet Cells; Humans; Hyperplasia; Hypersensitivity; Interleukin-13; Lung; Lung Neoplasms; Mice; Mice, Knockout; Mucins; Mucus; Ovalbumin; Phosphoproteins; Pulmonary Disease, Chronic Obstructive; Respiratory Mucosa; STAT6 Transcription Factor; TYK2 Kinase

Allergic lung inflammation is mediated by allergen-specific T responses, which are negatively regulated by regulatory T cells (Tregs). Previous studies have reported that inoculation of indigenous Clostridium species in the early lives of mice can induce Tregs that colonize the colon. However, whether inoculation of C leptum alone in adult mice could induce systemic Treg responses and inhibit allergic airway inflammation remains unclear.. To investigate the effect of oral administration of C leptum on systemic Treg responses and allergic airway inflammation in a mouse model of asthma.. Adult BABL/c mice were injected with ovalbumin to induce asthma and treated orally with C leptum or vehicle daily for 2 weeks. The numbers of Foxp3(+)CD4(+)CD25(+) Tregs in both the spleen and mediastinal lymph nodes were examined by flow cytometry. After allergen challenge, the airway hyperresponsiveness of individual mice was measured, and the numbers of inflammatory infiltrates and the levels of cytokines in bronchoalveolar lavage fluids ere determined.. Oral feeding with C leptum increased the percentage and total number of Tregs in the spleens and mediastinal lymph nodes at 14 days after inoculation and attenuated allergen-induced airway hyperresponsiveness and inflammation by inhibiting inflammatory cytokine production but enhancing interleukin 10 and transforming growth factor β1 production in the lungs.. Oral treatment with C leptum can attenuate induced allergic airway inflammation in adult mice.

Topics: Administration, Oral; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Clostridium; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

Allergic asthma is associated with excessive T helper type 2 (Th2) cells activation and airway hyperreactivity (AHR), implicated in the context of significant morbidity and mortality. Soluble ST2, a member of the interleukin (IL)-1 receptor family, has been shown to play a critical role in modulation of inflammatory disorders, yet the function of soluble ST2 in allergic inflammation remains unclear. In this study, we examined the possibility of regulating ovalbumin (OVA)-challenged airway inflammation by recombinant adenovirus-mediated sST2-Fc (Ad-sST2-Fc) gene transfer. Single intranasal administration of Ad-sST2-Fc before allergen challenge in OVA-immunized mice profoundly reduced serum immunoglobulin (Ig)E secretion, eosinophil infiltration and concentrations of IL-4, IL-5 and IL-13 in bronchoalveolar lavage fluid compared with administration of a control Ad vector. Histopathological examination of the lungs revealed that sST2-Fc over-expression markedly suppressed allergen-induced peribronchial inflammation and disruption of the alveolar architecture. Moreover, the beneficial effect of sST2-Fc in allergic lung inflammation is related to blocking the IL-33/ST2L signalling. Taken together, these results suggested that administration of Ad-sST2-Fc gene transfer may have therapeutic potential for the immunomodulatory treatment of OVA-mediated allergic pulmonary diseases.

Topics: Adenoviridae; Administration, Intranasal; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Genetic Therapy; Genetic Vectors; Humans; Hypersensitivity; Immunoglobulin E; Interleukin-1 Receptor-Like 1 Protein; Interleukin-13; Interleukin-33; Interleukin-4; Interleukin-5; Interleukins; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Interleukin; Receptors, Interleukin-1; Recombinant Fusion Proteins; Th2 Cells

The mechanism by which many monosensitized allergic individuals progress to polysensitization over time remains to be elucidated. Mouse models have contributed greatly to the understanding of sensitization to inhaled allergens in healthy airways but hardly any studies have addressed sensitization during established allergy. We hypothesized that an allergic inflammatory milieu might facilitate sensitization to inhaled allergens by the presence of mature dendritic cells (DCs) and IL-4.. Mice with house dust mite (HDM)-induced allergic airway inflammation received a single intratracheal dose of ovalbumin (OVA), 2 days after the last HDM exposure. Ten days later, sensitization was assessed by rechallenge with OVA. We evaluated the following factors for their importance in neosensitization: (1) maturation and recruitment of DCs to the airways, (2) dependency on DCs using CD11cDTR conditional knockout mice, (3) presence of ongoing airway inflammation by comparing sensitization at day 2 and day 14 after the last HDM exposure and (4) dependency on IL-4 by treatment with blocking antibodies.. House dust mite -induced inflammation facilitated neosensitization to OVA. HDM-induced inflammation increased the number of airway DCs with a mature phenotype but a DC reduction of 93% did not inhibit sensitization. Neosensitization to OVA was dependent on ongoing inflammation and in particular on IL-4.. These findings show that HDM-induced allergic airway inflammation facilitates neosensitization to a second inhaled allergen in an IL-4-dependent manner and provide insight into the underlying mechanism of the frequently observed progression to polysensitization in HDM-monosensitized individuals.

Topics: Administration, Inhalation; Allergens; Animals; Asthma; CD11c Antigen; Dendritic Cells; Female; Hypersensitivity; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Pyroglyphidae

Airway hyperreactivity is characterized by increased responsiveness to bronchoconstrictor stimuli and it is hallmark of asthma. Adenosine is an ubiquitous signaling nucleoside resulting from ATP catabolism, whose extracellular levels increase following cellular damage or stress. Adenosine plays a role in asthma; asthmatics, but not normal subjects, present bronchoconstriction following inhalation of adenosine or of its precursor, adenosine-5'-monophosphate, most likely via adenosine A(2B) receptor on mast cells. However, the mechanism underling the increased airway smooth muscle sensitivity to adenosine in asthmatics remains to be elucidated. Early experimental studies suggested the involvement of A(1) receptor; this hypothesis has been confirmed by more recent studies on guinea pigs and is corroborated by the finding of an increased adenosine A(1) expression on asthmatic bronchial tissues. Brown Norway rats, the strain usually used to assess asthma models, develop hyperresponsiveness to adenosine 3h following allergen challenge, but not 24h thereafter, without involvement of A(1) receptor. Here, we investigated the role of adenosine A(1) receptor in sensitized Wistar rats showing airway hyperresponsiveness 24h following allergen challenge. We found that on bronchi of sensitized Wistar rats challenged with allergen there is an increased adenosine A(1) receptor expression on smooth muscle that is responsible for hyperresponsiveness to adenosine and ovalbumin.

Topics: Adenosine; Adenosine A1 Receptor Antagonists; Allergens; Animals; Bronchi; Edema; Gene Expression Regulation; Hypersensitivity; Immunization; Male; Ovalbumin; Rats; Rats, Wistar; Receptor, Adenosine A1; Time Factors

The facilitating effects of multiwalled carbon nanotubes (MWCNT) on allergic asthma have not been sufficiently examined, although MWCNT appear to significantly increase the risk of health problems from occupational or environmental exposure. In this study, we examined whether sensitization by the combination of MWCNT with ovalbumin (OVA) promotes allergic asthmatic responses. BALB/c mice administered vehicle, MWCNT, OVA, or MWCNT+OVA through an intranasal route were challenged with OVA intratracheally four times. In the MWCNT+OVA group, the fourth challenge caused not only early- but also late-phase increases in airway resistance, although these responses were not observed in the vehicle, MWCNT, or OVA group; furthermore, the extents of the early and late responses were comparable to those in mice systemically sensitized with OVA+alum. Sensitization with MWCNT and OVA promoted airway inflammation and goblet cell hyperplasia in the lung compared with the vehicle, MWCNT or OVA group. In addition, adjuvant activity for OVA-specific immunoglobulin E (IgE), IgG1, and IgG2a production in serum and increased levels of interleukin-4 (IL-4), IL-5, IL-13, and IL-17 in the lung tissue were observed. In conclusion, these results suggest that exposure to MWCNT and antigen can induce a biphasic increase in airway resistance, airway inflammation, goblet cell hyperplasia, and the production of antigen-specific antibodies. This study highlights the risk of exposure to a combination of MWCNT with antigen.

Topics: Airway Resistance; Allergens; Alum Compounds; Animals; Asthma; Goblet Cells; Hyperplasia; Hypersensitivity; Immunoglobulins; Inflammation; Interleukins; Lung; Male; Mice; Mice, Inbred BALB C; Nanotubes, Carbon; Ovalbumin

Angiopoietin (Ang)1 and Ang2 are ligands for Tie2 tyrosine kinase receptor (Tie2). Elevated levels of Ang1 and Ang2 in induced sputum of patients with asthma have been reported, with a positive correlation of Ang2 levels with the severity of airway occlusion. Although studies have shown Tie2-mediated regulation of nonvascular cells in some pathological conditions, current knowledge on Tie2 signaling in asthma is limited to the vasculature. We examined the expression pattern of Ang1, Ang2, vascular endothelial growth factor (VEGF), and Tie2 and their correlation with the degree of airway remodeling in the lung of ovalbumin (OVA)-sensitized and OVA-challenged mice with airway hyperresponsiveness. Lung tissues were isolated from Balb/c mice after OVA sensitization and challenge. Hematoxylin and eosin, periodic acid-Schiff, and trichrome staining were used to show the lung pathology. The expression of Ang1, Ang2, VEGF, and Tie2 was examined using immunofluorescence, Western blot, ELISA, and real-time PCR. In the lung of normal mice, Tie2 expression was detected only in the blood vessels. However, in the lung of OVA-sensitized and OVA-challenged mice, Tie2 was abundantly expressed in airway epithelial cells and in a subset of macrophages in addition to constitutive expression in pulmonary vessels. The increase in Tie2 expression correlated with the severity of airway remodeling. Macrophages and airway epithelial cells express Ang2 and VEGF only in allergic models. Ang1 was constitutively expressed, with a decrease in mRNA level in allergic models. In conclusion, increased expression of Tie2 and Ang2 in allergic airway epithelium and alveolar macrophages correlates with the severity of airway remodeling.

Topics: Angiopoietin-1; Angiopoietin-2; Animals; Asthma; Gene Expression Regulation; Hypersensitivity; Inflammation; Macrophages; Methacholine Chloride; Mice; Mice, Inbred BALB C; Microscopy, Fluorescence; Models, Biological; Ovalbumin; Receptor Protein-Tyrosine Kinases; Receptor, TIE-2; Vascular Endothelial Growth Factor A

Induction of nitric oxide synthase (NOS)-2 and production of nitric oxide (NO) are common features of allergic airway disease. Conditions of severe asthma are associated with deficiency of airway S-nitrosothiols, a biological product of NO that can suppress inflammation by S-nitrosylation of the proinflammatory transcription factor, NF-κB. Therefore, restoration of airway S-nitrosothiols might have therapeutic benefit, and this was tested in a mouse model of ovalbumin (OVA)-induced allergic inflammation. Naive or OVA-sensitized animals were administered S-nitrosoglutathione (GSNO; 50 μl, 10 mM) intratracheally before OVA challenge and analyzed 48 hours later. GSNO administration enhanced lung tissue S-nitrosothiol levels and reduced NF-κB activity in OVA-challenged animals compared with control animals, but did not lead to significant changes in total bronchoalveolar lavage cell counts, differentials, or mucus metaplasia markers. Administration of GSNO also altered the activation of hypoxia-inducible factor (HIF)-1, leading to HIF-1 activation in naive mice, but suppressed HIF-1 activation in OVA-challenged mice. We assessed the contribution of endogenous NOS2 in regulating NF-κB and/or HIF-1 activation and allergic airway inflammation using NOS2(-/-) mice. Although OVA-induced NF-κB activation was slightly increased in NOS2(-/-) mice, associated with small increases in bronchoalveolar lavage neutrophils, other markers of allergic inflammation and HIF-1 activation were similar in NOS2(-/-) and wild-type mice. Collectively, our studies indicate that instillation of GSNO can suppress NF-κB activation during allergic airway inflammation, but does not significantly affect overall markers of inflammation or mucus metaplasia, thus potentially limiting its therapeutic potential due to effects on additional signaling pathways, such as HIF-1.

Topics: Animals; Bronchoalveolar Lavage Fluid; Hypersensitivity; Hypoxia-Inducible Factor 1; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; NF-kappa B; Nitric Oxide; Nitric Oxide Donors; Ovalbumin; S-Nitrosoglutathione

Transglutaminase 2 (TGase 2) expression is increased in inflammatory diseases, and TGase 2 inhibitors block these increases. We examined whether the R2 peptide inhibited the expression of TGase 2 in a mouse model of inflammatory allergic asthma.. C57BL/6 mice were sensitized and challenged by ovalbumin (OVA) to induce asthma. OVA-specific serum IgE and leukotrienes (LTs) levels were measured by enzyme-linked immunosorbent assay. Recruitment of inflammatory cells into bronchoalveolar lavage (BAL) fluid or lung tissues and goblet cell hyperplasia were assessed histologically. Airway hyperresponsiveness was determined in a barometric plethysmographic chamber. Expression of TGase 2, eosinophil major basic protein (EMBP), the adhesion molecule vascular cell adhesion molecule-1, Muc5ac and phospholipase A(2) (PLA(2) ) protein were determined by Western blot. Expression of mRNAs of Muc5ac, cytokines, matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) were measured by reverse transcriptase-polymerase chain reaction and nuclear factor-κB (NF-κB) by electrophoretic mobility shift assay.. R2 peptide reduced OVA-specific IgE levels; the number of total inflammatory cells, macrophages, neutrophils, lymphocytes and eosinophils in BAL fluid and the number of goblet cells. Airway hyperresponsiveness, TGase 2 and EMBP levels, mRNA levels of interleukin (IL)-4, IL-5, IL-6, IL-8, IL-13, RANTES, tumour necrosis factor-α, and MMP2/9, Muc5ac, NF-κB activity, PLA(2) activity and expressions, and LT levels in BAL cells and lung tissues were all reduced by R2 peptide. R2 peptide also restored expression of TIMP1/2.. R2 peptide reduced allergic responses by regulating NF-κB/TGase 2 activity in a mouse model of allergic asthma. This peptide may be useful in the treatment of allergic asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Base Sequence; Blotting, Western; Bronchoalveolar Lavage Fluid; Disease Models, Animal; DNA Primers; Enzyme Inhibitors; Female; GTP-Binding Proteins; Hypersensitivity; Mice; Mice, Inbred C57BL; Ovalbumin; Peptides; Protein Glutamine gamma Glutamyltransferase 2; Reverse Transcriptase Polymerase Chain Reaction; Transglutaminases

Increasing evidence indicates that eosinophils contribute greatly to airway remodeling in asthma. Since interleukin-5 (IL-5) plays a critical role in the regulation of eosinophils in asthma, anti-IL-5 therapy may be a novel approach to inhibit airway remodeling in asthma.. In this study, we applied a recombinant adeno-associated virus vector-mediated antisense IL-5 gene delivery (rAAV-ASIL-5) system to investigate its effect on airway remodeling in ovalbumin (OVA)-sensitized and -challenged rats.. rAAV-ASIL-5 was used to infect OVA-sensitized and -challenged rats. IL-5 protein in bronchoalveolar lavage fluid (BALF) was detected by ELISA. The eosinophils in BALF were counted. Transforming growth factor (TGF)-β1- and TGF-β2-positive cells in the peribronchial space were detected by immunohistochemical staining. Lung tissue was collected for Sirius red staining and histological analysis.. rAAV-ASIL-5 significantly decreased the level of IL-5 protein, the number of eosinophils in BALF and the numbers of TGF-β1- and TGF-β2-positive cells in the peribronchial space. The area of Sirius red staining in airways was also decreased. Moreover, the rAAV-ASIL-5 treatment inhibited the increase in total bronchial wall area and airway smooth muscle area.. These results suggest that rAAV-ASIL-5-based gene therapy could be used to inhibit airway remodeling in allergic rats.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Dependovirus; DNA, Antisense; Eosinophils; Genetic Therapy; Genetic Vectors; Humans; Hypersensitivity; Interleukin-5; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; Recombination, Genetic; Treatment Outcome

Although numerous studies demonstrate the participation of nitric oxide (NO) in various inflammatory diseases, the precise function of NO in allergic asthma remains unclear. We investigated whether iNOS inhibition could interfere with the kinetics of VLA-4 and Mac-1 expression and adhesion properties of bone marrow and peripheral blood eosinophils of sensitized mice after antigen exposure. Treatment of allergic mice with 1400 W (iNOS inhibitor) increased the adhesion of bone marrow eosinophils to ICAM-1, but not blood eosinophils, at 24h and 48 h after OVA-challenge. Conversely, adhesion of blood eosinophils from 1400 W-treated mice to VCAM-1 diminished at 24h and was almost completely blocked at 48 h. 1400 W did not induce any change in the adhesion of bone marrow eosinophils to VCAM-1, at 24h, but cells collected 48 h after challenge showed significantly lower adherence. Flow cytometry demonstrated that 1400 W resulted in a significantly increased Mac-1 expression on bone marrow eosinophils at 24h, as compared to control mice. However, at 24h, 1400 W significantly decreased Mac-1 and VLA-4 expressions on blood eosinophils. At 48 h, the expressions of both Mac-1 and VLA-4 returned to previous levels. Results show a temporal effect of iNOS upon Mac-1 expression and function, the chief adhesion molecule involved in the eosinophil efflux from the bone marrow at 24h. In contrast, Mac-1 and VLA-4 were involved in eosinophil mobilization from blood to lungs at 48 h after antigen challenge. Data suggest an important role of the Mac-1 and VLA-4 in the iNOS-modulated migration of eosinophils to the lungs of allergic mice.

Topics: Amidines; Animals; Benzylamines; Bone Marrow; Chemotaxis, Leukocyte; Cytokines; Disease Models, Animal; Enzyme Inhibitors; Eosinophils; Female; Hypersensitivity; Integrin alpha4beta1; Leukocyte Count; Lung; Macrophage-1 Antigen; Mice; Mice, Inbred BALB C; Nitric Oxide; Nitric Oxide Synthase Type II; Ovalbumin; Th2 Cells

  Tumor necrosis factor (TNF)-α is a principal mediator of the acute inflammatory response, including allergic rhinitis. TNF-α inhibitors are widely used for the treatment of inflammatory conditions such as rheumatoid arthritis and inflammatory bowel diseases; however, the effects of TNF-α inhibitors on allergic rhinitis are not well established. We aimed to investigate the effects of infliximab, a TNF-α inhibitor, on allergic rhinitis in a mouse model..   BALB/c mice were sensitized with ovalbumin (OVA) and alum, and challenged intranasally with OVA. The TNF-α inhibitor, infliximab was administered intraperitoneally, and multiple parameters of allergic responses were evaluated to determine the effects of infliximab..   Infliximab reduced allergic symptoms and eosinophilic infiltration into the nasal mucosa. It also suppressed total and OVA-specific IgE levels, and inhibited local Th2 cytokine transcription in the nasal mucosa and systemic Th2 cytokine production by splenocytes. Furthermore, the expression of E-selectin, neither intercellular adhesion molecule 1 (ICAM-1) nor vascular cell adhesion molecule 1 (VCAM-1), in the nasal mucosa was suppressed in the infliximab-treated group when compared to the nontreated group..   This study shows that the TNF-α inhibitor infliximab induces anti-allergic effects by decreasing local and systemic Th2 cytokine (IL-4) production, total and OVA-specific IgE levels, adhesion molecule (E-selectin) expression, and eosinophil infiltration into the nasal mucosa in an allergic rhinitis model. Therefore, infliximab should be considered as a potential agent in treating allergic rhinitis.

Topics: Animals; Anti-Allergic Agents; Antibodies, Monoclonal; Cell Adhesion Molecules; Cytokines; Eosinophils; Hypersensitivity; Immunoglobulin E; Infliximab; Mice; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Rhinitis; Th2 Cells

A transgenic papaya line (TPY10-4) that is resistant to both papaya ringspot virus (PRSV) and papaya leaf distortion mosaic virus (PLDMV) has been developed in Taiwan. This study investigated the immunomodulatory properties of transgenic TPY10-4 and its native (TCK) papaya fruits using an ovalbumin (OVA)-sensitised mouse model. Both green and ripe papaya fruits at low (0.2 g powder kg(-1) body weight (BW)) and high (1.6 g powder kg(-1) BW) doses were administered to experimental mice by intragastric gavage for 5 weeks. Changes in serum total immunoglobulin A (IgA), IgE, IgG and IgM levels, OVA-specific IgE, IgG1 and IgG2a titres and Th1/Th2 cytokine secretions using splenocytes were determined.. Transgenic TPY10-4 or native TCK papaya fruit supplementation did not significantly affect body, visceral organ and relative tissue weights, total IgE antibody levels, OVA-specific IgE and IgG1 antibody titres or OVA-stimulated interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4, IL-5 and IL-10 secretions using splenocytes. However, transgenic papaya fruits markedly increased serum total IgM levels.. This study suggests that transgenic TPY10-4 papaya fruits do not increase the allergenic potential of OVA by oral administration but may have a protective immunity via increasing the serum total IgM level.

Topics: Administration, Oral; Animals; Carica; Dietary Supplements; Female; Food, Genetically Modified; Fruit; Hypersensitivity; Immunity; Immunoglobulin M; Immunologic Factors; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Plant Preparations; Plants, Genetically Modified; Spleen

Epidemiologic studies have associated higher dietary consumption of soy isoflavones with decreased self-report of cough and allergic respiratory symptoms, but the pharmacodynamic effects of soy isoflavone on asthmatic model have not been well-described. Here, we hypothesized that soy isoflavone may have potential effects on airway hyperresponsiveness, inflammation and airway remodeling in a murine of asthma. Mice sensitized and challenged with ovalbumin developed airway inflammation. Bronchoalveolar lavage fluid was assessed for inflammatory cell counts, and for cytokine levels. Lung tissues were examined for cell infiltration, mucus hypersecretion and airway remodeling, and for the expression of inflammatory biomarkers. Airway hyperresponsiveness was monitored by direct airway resistance analysis. Oral administration of soy isoflavone significantly reduced ovalbumin-induced airway hyperresponsiveness to intravenous methacholine, and inhibited ovalbumin-induced increases in eosinophil counts. RT-PCR analysis of whole lung lysates revealed that soy isoflavone markedly suppressed ovalbumin-induced mRNA expression of eotaxin, interleukin(IL)-5, IL-4 and matrix metalloproteinase-9, and increased mRNA expression of interferon (IFN)-γ and tissue inhibitor of metalloproteinase-1 in a dose-dependent manner. Soy isoflavone also substantially recovered IFN-γ/IL-4 (Th1/Th2) levels in bronchoalveolar lavage fluid. In addition, histologic studies showed that soy isoflavone dramatically inhibited ovalbumin-induced lung tissue eosinophil infiltration, airway mucus production and collagen deposition in lung tissues. Our findings suggest that soy isoflavone as nutritional supplement may provide a novel means for the treatment of airway inflammatory disease.

Topics: Airway Remodeling; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Collagen; Eosinophils; Female; Glycine max; Hypersensitivity; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-5; Isoflavones; Lung; Matrix Metalloproteinase 9; Methacholine Chloride; Mice; Mice, Inbred ICR; Mucus; Ovalbumin; Peroxidase; Respiratory System; Superoxide Dismutase; Tissue Inhibitor of Metalloproteinase-1

Mast cells stimulation activates degranulation process resulting in releasing of mediators, such as histamine. In this study, the effect of aqueous extract of sitagliptin, a selective dipeptidylpeptidase-4 inhibitor, on the mast cell-mediated allergic response was studied with the possible mechanisms of action, focusing on the histamine release and pro-inflammatory cytokine secretion in mast cells. Sitagliptin produced dose dependent inhibition in compound 48/80-induced systemic reactions. In addition, sitagliptin attenuated IgE-mediated skin allergic reaction. Sitagliptin dose-dependently reduced compound 48/80- and IgE-induced histamine release from mast cells. Sitagliptin decreased the secretion of pro-inflammatory cytokines, tumor necrosis factor-α, in mast cells. So, the finding of this study provides evidence that sitagliptin inhibits mast cell derived allergic reactions, and involvement of pro-inflammatory cytokine secretion in such effects.

Topics: Anaphylaxis; Animals; Cytokines; Dose-Response Relationship, Drug; Histamine Release; Hypersensitivity; Immunoglobulin E; Male; Mast Cells; Mice; Ovalbumin; p-Methoxy-N-methylphenethylamine; Passive Cutaneous Anaphylaxis; Pyrazines; Rats; Rats, Wistar; Sitagliptin Phosphate; Triazoles; Tumor Necrosis Factor-alpha

Epidemiological and experimental evidence supports the notion that microbial infections that are known to induce Th1-type immune responses can suppress Th2 immune responses, which are characteristics of allergic disorders. However, live microbial immunization might not be feasible for human immunotherapy. Here, we evaluated whether induction of Th1 immunity by the immunostimulatory sequences of CpG-oligodeoxynucleotides (CpG-ODN), with or without culture filtrate proteins (CFP), from Mycobacterium tuberculosis would suppress ongoing allergic lung disease. Presensitized and ovalbumin (OVA)-challenged mice were treated subcutaneously with CpG, or CpG in combination with CFP (CpG/CFP). After 15 days of treatment, airway inflammation and specific T- and B-cell responses were determined. Cell transfer experiments were also performed. CpG treatment attenuated airway allergic disease; however, the combination CpG/CFP treatment was significantly more effective in decreasing airway hyperresponsiveness, eosinophilia and Th2 response. When an additional intranasal dose of CFP was given, allergy was even more attenuated. The CpG/CFP therapy also reduced allergen-specific IgG1 and IgE antibodies and increased IgG2a. Transfer of spleen cells from mice immunized with CpG/CFP also reduced allergic lung inflammation. CpG/CFP treatment induced CFP-specific production of IFN-γ and IL-10 by spleen cells and increased production of IFN-γ in response to OVA. The essential role of IFN-γ for the therapeutic effect of CpG/CFP was evidenced in IFN-γ knockout mice. These results show that CpG/CFP treatment reverses established Th2 allergic responses by an IFN-γ-dependent mechanism that seems to act both locally in the lung and systemically to decrease allergen-specific Th2 responses.

Topics: Adoptive Transfer; Animals; Antigens, Bacterial; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophilia; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Interferon-gamma; Interleukin-10; Lung Diseases; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Mycobacterium tuberculosis; Oligodeoxyribonucleotides; Ovalbumin; Spleen; Th1 Cells; Th2 Cells

The calcium-binding protein S100A12 is highly up-regulated in the serum and sputum of patients with allergic asthma and is suggested to be a biomarker and pathologic mediator of asthma.. To test the role of S100A12 in mediating airway inflammation in a mouse model of allergic lung inflammation.. Transgenic (TG) mice that express human S100A12 and wild-type (WT) littermates were sensitized and challenged with ovalbumin (OVA) and assessed for inflammation, lung structure, and function.. Following OVA sensitization and challenge, S100A12 TG mice showed reduced peribronchial and perivascular inflammation, mucus production, and eosinophilia as well as attenuated airway responsiveness to contractile agonist compared with WT sensitized and challenged animals. This is explained, at least in part, by remodelled airways in S100A12 TG mice with thinning of the airway smooth muscle. S100A12 exposure induced Fas expression and activation of caspase 3 in cultured airway smooth muscle cells, suggesting that airway smooth muscle abnormalities observed in S100A12 TG mice may be mediated through myocyte apoptosis.. S100A12 is one of the most abundant proteins found in the airways of human asthmatics, and it was postulated that S100A12 could mediate the inflammatory process. Our study shows for the first time that TG expression of S100A12 in the lung of mice does not exacerbate lung inflammation in a model of OVA-induced allergic inflammation. We speculate that the high levels of S100/calgranulins found in bronchoalveolar lavage fluid of asthmatics and of OVA-treated TG S100A12 mice do not significantly mediate pulmonary inflammation.

Topics: Airway Remodeling; Animals; Apoptosis; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Chemokines; Disease Models, Animal; Humans; Hypersensitivity; Immunity, Humoral; Mice; Mice, Inbred C57BL; Mice, Transgenic; Myocytes, Smooth Muscle; Ovalbumin; Pneumonia; Respiratory Mucosa; Respiratory System; S100 Proteins; S100A12 Protein

Epicutaneous immunotherapy onto intact skin has proved to be an efficient and safe alternative treatment of allergy in an animal model with various allergens and in children for cow's milk allergy. The aim of this study was to analyze the different steps of the immunological handling of the allergen when deposited on intact skin using an epicutaneous delivery system and its immune consequences in sensitized BALB/c mice. As expected, when applied on intact skin, OVA exhibits neither a passive passage through the skin nor any detectable systemic delivery. The current study demonstrates that, after a prolonged application on intact skin, OVA is taken up by dendritic cells in the superficial layers of the stratum corneum and transported, after internalization, to the draining lymph nodes, with variations according to the previous level of sensitization of the mice. When OVA is applied with the epicutaneous delivery system repeatedly, specific local and systemic responses are down-modulated in association with the induction of regulatory T cells. Besides providing new insights into skin function in the presence of allergens, this study indicates that the skin might have a tolerogenic role, at least when kept intact.

Topics: Administration, Cutaneous; Allergens; Animals; Cell Movement; Dendritic Cells; Desensitization, Immunologic; Female; Hypersensitivity; Immune Tolerance; Immunotherapy; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Skin; T-Lymphocytes, Regulatory

Lactobacilli isolated from Kimchi, a Korean traditional food, were tested for their capacity to modulate the T helper (Th) 1/Th2 balance. Ovalbumin (OVA)-sensitized mouse splenocytes were cultured with 26 strains of lactobacilli; the highest IL-12 induction and lowest IL-4 production were then observed in 4 strains, including Lactobacillus plantarum CJLP55, CJLP56, CJLP133, and CJLP136. These strains produced a larger amount of IL-12, which enhances differentiation and activation of Th1 cells, in macrophage cell-lines more than positive control strains L. casei KCTC 3109(T) and L. rhamnosus GG, although they also induced production of IL-10, which is a suppressor of IL-12. Indeed, CJLP133-stimulated macrophages induced production of more Th1 cytokine IFN-γ and less Th2 cytokine IL-4 than KCTC 3109(T) and GG in co-cultivation with T cells. These findings suggest that lactobacilli from Kimchi may modulate the Th1/Th2 balance via macrophage activation in the hypersensitive reaction caused by Th2 cells.. Allergic reactions including asthma and atopy are caused by predominance of Th2 response over Th1 response. Lactobacilli isolated from fermented foods such as yogurt, cheese, and Kimchi showed health-promoting activities. The present study indicated that several lactobacilli strains from Kimchi may reduce allergic reactions through macrophage-mediated induction of Th1 response.

Topics: Animals; Cell Line; Fermentation; Food Microbiology; Hypersensitivity; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Lactobacillus; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th1-Th2 Balance; Th2 Cells

The activation of C-fibers in the airways induces coughing, mucus production and bronchoconstriction, which are also symptoms of airway diseases. In this study, we evaluated the role of the C-fibers and the TRPV1 (transient receptor potential vanilloid 1) receptor in an experimental mouse model of allergic airway inflammation. To study the role of C-fibers, we either degenerated the C-fibers persistently (capsaicin administration in neonate mice) or transiently (capsaicin administration in adult mice). No alteration was observed in eosinophil recruitment to the bronchoalveolar lavage fluid in animals treated with capsaicin in the neonatal period. However, in adult animals, capsaicin treatment after the first ovalbumin challenge (in the establishment of the inflammatory process) decreased the eosinophil numbers. This effect was more pronounced in adult animals treated with capsaicin before beginning the ovalbumin immunization (in the development of the inflammatory process). In addition, interleukin (IL)-5 and chemokine ligand 11 (CCL11) levels in the bronchoalveolar lavage fluid, as well as P-selectin expression and p65 nuclear factor κB (NF-κB) activation in the lung were also decreased. No alterations were observed in the IL-10 and IL-13 levels. Next we determined the effect of TRPV1 receptor blockade on allergic airway inflammation. SB366791 administrated in mice by intraperitoneal (500μg/kg) or intranasal (0.1, 1 or 10nmol/site) route failed to decrease eosinophil recruitment to the bronchoalveolar lavage fluid or alter any other metrics cited above. Thus, the present results confirm and extend previous data supporting the involvement of C-fibers, but not the TRPV1 receptor, in allergic airway inflammation.

Topics: Allergens; Anilides; Animals; Animals, Newborn; Bronchoalveolar Lavage Fluid; Capsaicin; Cell Count; Cinnamates; Cytokines; Female; Gene Expression Regulation; Hypersensitivity; Immunization; Inflammation; Leukocytes; Lung; Mice; Mice, Inbred BALB C; Nerve Fibers, Unmyelinated; NF-kappa B; Ovalbumin; P-Selectin; Respiratory System; TRPV Cation Channels

Some epidemiologic studies have indicated that attendance to chlorinated swimming pools is associated with airway hyperreactivity (AHR), allergies and asthma.. To investigate the effects of sodium hypochlorite (NaClO), the main pool disinfectant, on allergic sensitization and airway inflammation in mice.. In a first series of experiments, mice were sensitized to ovalbumin (OVA), followed by OVA aerosols with or without prior nasal instillation of NaClO (3ppm active chlorine). In a second series, naïve mice received 1-7 nasal instillations of OVA, 10min after instillations of NaClO or water. After 1, 3, 5 and 7 exposures airway reactivity to methacholine, cellular inflammation in bronchoalveolar lavage (BAL), serum OVA-specific IgEs and lung Th2 cytokines were measured.. In the first mouse model, airway allergy parameters were not significantly altered upon NaClO administration. However in the second model, NaClO exposure prior to OVA did induce AHR, already after 1 combined application. Combined NaClO+OVA exposure did not lead to an influx of inflammatory cells in BAL fluid or production of anti-OVA IgEs. No AHR developed when OVA was heat-denatured, pre-chlorinated, or replaced by bovine serum albumin or lipopolysaccharide.. Nasal instillation of NaClO prior to OVA induces AHR without allergic sensitization. This response is OVA-specific.

Topics: Animals; Bronchial Hyperreactivity; Hypersensitivity; Hypochlorous Acid; Male; Mice; Mice, Inbred BALB C; Ovalbumin

The flowers of Inula japonica (Inulae Flos) have long been used in traditional medicine for treating inflammatory diseases. The effects on OVA-induced asthmatic mice of an Inulae Flos extract (IFE) were evaluated in this study. The anti-asthmatic effects of IFE were determined by observing eosinophil recruitment, airway hyper-responsiveness (AHR), Th2 cytokine and IgE levels, and lung histopathology. The IFE treatment effectively reduced the percentage of eosinophils and Th2 cytokines in the bronchoalveolar lavage fluid (BALF) when compared to the levels in OVA-induced mice. IFE also suppressed AHR induced by aerosolized methacholine in OVA-induced mice. The results of the histopathological studies indicate that inflammatory cell infiltration and mucus hypersecretion were both inhibited by the IFE administration when compared to the effect on OVA-induced mice. The IFE treatment also suppressed the serum IgE levels and decreased Th2 cytokines in the supernatant of cultured splenocytes. These results suggest that IFE may have therapeutic potential against asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Female; Flowers; Hypersensitivity; Immunoglobulin E; Inflammation; Inula; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Spleen; T-Lymphocytes

In this study, we evaluated the anti-inflammatory response and the mechanism by which dehydroepiandrosterone modulates immunity in ovalbumin-sensitized asthmatic mice. Female BALB/c mice were sensitized and challenged with ovalbumin and then treated with oral administration of dehydroepiandrosterone on days 21 to 27. The results showed dehydroepiandrosterone could suppress airway hyperresponsiveness and decrease eosinophil infiltration of the lungs in ovalbumin-sensitized mice. Moreover, dehydroepiandrosterone inhibited chemokines, including CCL11/eotaxin-1 and CCL24/eotaxin-2, and Th2-associated cytokine levels in bronchoalveolar lavage fluid. After the inflammatory human bronchial epithelial cell line BEAS-2B was treated with dehydroepiandrosterone, levels of proinflammatory cytokines and chemokines were inhibited, including IL-6, IL-8, CCL11, and CCL24. We suggested that dehydroepiandrosterone inhibited inflammation in bronchial epithelial cells as indicated by the suppression of Th2-associated cytokines and chemokines. Dehydroepiandrosterone also suppressed eosinophil migration and infiltration into the lung to improve the symptoms of asthma in ovalbumin-sensitized mice.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Movement; Cells, Cultured; Chemokines; Cytokines; Dehydroepiandrosterone; Dinoprostone; Eosinophils; Epithelial Cells; Female; Humans; Hypersensitivity; Intercellular Adhesion Molecule-1; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory System; Spleen; Th2 Cells

Phosphoinositide 3-kinase (PI3K)/Akt pathway is linked to the development of asthma. Anti-malarial drug artesunate is a semi-synthetic derivative of artemisinin, the principal active component of a medicinal plant Artemisia annua, and has been shown to inhibit PI3K/Akt activity. We hypothesized that artesunate may attenuate allergic asthma via inhibition of the PI3K/Akt signaling pathway.. Female BALB/c mice sensitized and challenged with ovalbumin (OVA) developed airway inflammation. Bronchoalveolar lavage fluid was assessed for total and differential cell counts, and cytokine and chemokine levels. Lung tissues were examined for cell infiltration and mucus hypersecretion, and the expression of inflammatory biomarkers. Airway hyperresponsiveness was monitored by direct airway resistance analysis. Artesunate dose-dependently inhibited OVA-induced increases in total and eosinophil counts, IL-4, IL-5, IL-13 and eotaxin levels in bronchoalveolar lavage fluid. It attenuated OVA-induced lung tissue eosinophilia and airway mucus production, mRNA expression of E-selectin, IL-17, IL-33 and Muc5ac in lung tissues, and airway hyperresponsiveness to methacholine. In normal human bronchial epithelial cells, artesunate blocked epidermal growth factor-induced phosphorylation of Akt and its downstream substrates tuberin, p70S6 kinase and 4E-binding protein 1, and transactivation of NF-κB. Similarly, artesunate blocked the phosphorylation of Akt and its downstream substrates in lung tissues from OVA-challenged mice. Anti-inflammatory effect of artesunate was further confirmed in a house dust mite mouse asthma model.. Artesunate ameliorates experimental allergic airway inflammation probably via negative regulation of PI3K/Akt pathway and the downstream NF-κB activity. These findings provide a novel therapeutic value for artesunate in the treatment of allergic asthma.

Topics: Animals; Anti-Inflammatory Agents; Antimalarials; Artemisinins; Artesunate; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Enzyme Activation; Epidermal Growth Factor; Epithelial Cells; Female; Gene Expression Regulation; Humans; Hypersensitivity; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Pyroglyphidae; Signal Transduction; Th2 Cells

To investigate allergic reactions of traditional Chinese medicine (TCM) injections, and to determine the contents of serum IgE and histamine in sensitized animal. The correlation between the preceding contents in serum and allergic reactions may be found, thus offering experimental evidences for advancing the accuracy of anticipation by type I allergy.. We carried out passive cutaneous anaphylaxis (PCA) tests,active systemic anaphylaxis (ASA) tests and anaphylactoid reactions using three TCM injections, and determined the contents of serum OVA-sIgE, total serum IgE and histamine in sensitized animals by ELISA method.. The results of PCA test were negative, and there was no significant difference for total serum IgE level between experimental group and normal saline group. In the study of adjuvant effect in TCM injections + OVA (at the dose level that doesn't cause allergic reactions), the PCA results of SHL and YXC were positive and there was a increase in content of serum OVA-sIgE, while the PCA result of QKL was negative with a unobvious increase in the content of serum OVA-sIgE. The content of total serum IgE wasn't remarkably increased in each group and the results of ASA test were all positive. Three injections all caused anaphylactoid symptoms in guinea pigs in different doses or injection speed and the response intensity was found to be dosage and injection speed dependant. Furthermore, there was no significant difference for the content of total serum IgE in each group, whereas serum histamine concentration in every experimental group was markedly higher than normal saline group.. SHL and YXC increase the sensitivity of guinea pigs on OVA, and three TCM injections can cause allergic reactions in guinea pig. Allergic reactions of three TCM injections are correlated with specific IgE antibodies and histamine contents.

Topics: Animals; Female; Guinea Pigs; Histamine; Hypersensitivity; Immunoglobulin E; Injections; Male; Medicine, Chinese Traditional; Ovalbumin; Passive Cutaneous Anaphylaxis

We have previously shown that the allergic sensitization to ovalbumin does not represent a superantigen-like immune response. In gene-targeted mice (ΔD-iD) with a single modified Diversity gene segment (D(H)) of the immunoglobulin heavy chain, enriched for charged amino acids, the asthma phenotype in a murine model was markedly alleviated compared to wild-type animals.. We now sought to determine whether the confinement to a single D(H) gene segment alone leads to a reduced allergic phenotype.. We examined another gene-targeted mouse strain (ΔD-DFL) with a single D(H) gene segment which encodes for neutral amino acids, thus reflecting the preferential repertoire in wild-type mice. Mice were sensitized intraperitoneally to ovalbumin.. Despite the constraint to a single D(H) gene segment, ΔD-DFL mice mounted high total and allergen-specific IgG(1) and IgE serum levels after sensitization to ovalbumin. The affinity constants of allergen-specific IgG(1) antibodies did not differ between ΔD-DFL and wild type. Following challenge with aerosolized allergen, a marked local T(H)2 cytokine response and an eosinophilic airway inflammation developed. Quantitative histology revealed increased mucus production and intense goblet cell metaplasia which were identical to those in wild type. Moreover, ΔD-DFL mice developed an airway hyperreactivity to methacholine and to the specific allergen, which both did not differ from those in wild-type animals.. A single D(H) gene segment is sufficient for the establishment of the asthma phenotype in a murine model of allergic airway inflammation. Thus, the allergic phenotype depends on the amino acid composition and not on the diversity of the classical antigen-binding site.

Topics: Allergens; Animals; Asthma; B-Lymphocytes; Bronchial Hyperreactivity; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin

The syndrome of allergic asthma features reversible bronchoconstriction, airway inflammation and hyperresponsiveness as well as airway remodelling, including goblet cell hyperplasia. Managing severe asthma is still a clinical challenge. Numerous studies report that furosemide, an inhibitor of Na(+)-K(+)-Cl(-) cotransporter (NKCC) reduces airway hyperresponsiveness (AHR) in asthmatic patients. However, the mechanism by which furosemide exerts anti-asthmatic action remains unclear.. This study sought to investigate the cellular profile of NKCC1 expression in the lung and examine the effects of furosemide on several outcome measurements in a mouse model of allergic asthma.. Mice were sensitized and challenged with ovalbumin (OVA). Before challenge, the OVA-sensitized mice were treated with furosemide (4.0 mg/kg/day, via daily intraperitoneal injection for 5 days). Outcome measurements in naïve, OVA-exposure, furosemide-treated naïve and furosemide-treated OVA-exposed mice included the slope of the relationship between inhaled methacholine (MCh) concentration and respiratory system resistance (Slope·R(RS)), bronchoalveolar lavage (BAL) cell counts and immunohistochemical and immunoblotting assays of lung tissues.. NKCC1 immunoreactivity was observed in airway epithelial cells (AECs) and alveolar type II (ATII) cells of the control mice. OVA exposure enhanced the expression of NKCC1 in AECs and ATII cells, and increased the infiltration of NKCC1-expressing T lymphocytes in the lung. NKCC1 immunoreactivity was not detected in the airway smooth muscle (ASM) cells. Furosemide treatment reduced the Slope·R(RS) in both naïve and OVA-exposed mice by about 50%. Furosemide treatment also increased T lymphocyte infiltration to the lung in OVA-exposed mice by approximately 53%, but had no effect on pulmonary goblet cell hyperplasia.. Furosemide decreases basal airway responsiveness, thereby reducing the extent of allergen-induced AHR. However, the same treatment also increases T lymphocytes infiltration in the course of allergic asthma. Further studies are necessary to address the usefulness of furosemide in the clinical treatment of asthma.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Furosemide; Goblet Cells; Hypersensitivity; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Sodium-Potassium-Chloride Symporters; Solute Carrier Family 12, Member 2; T-Lymphocytes; Treatment Outcome

BALB/c mice received saline (SAL groups) or ovalbumin (OVA groups) intraperitoneally (days 1, 3, 5, 7, 9, 11 and 13). After 27 days, a burst of intratracheal OVA or SAL (days 40, 43 and 46) was performed. Animals were then divided into four groups (N=8, each) and intranasally instilled with saline (SAL-SAL and OVA-SAL) or residual oil fly ash (SAL-ROFA and OVA-ROFA). 24h later, total, initial and difference resistances (Rtot, Rinit, Rdiff) and static elastance (Est) were measured. Lung responsiveness to methacholine was assessed as slope and sensitivity of Est, Rtot, Rinit, and Rdiff. Lung morphometry (collapsed and normal areas and bronchoconstriction index) and cellularity (polymorphonuclear, mononuclear and mast cells) were determined. OVA or ROFA similarly impaired lung mechanics and increased the amount of polymorphonuclear cells and collapsed areas. OVA-ROFA showed even higher hyperresponsiveness, bronchoconstriction and mast cell infiltration. Thus, we concluded that ROFA exposure may add an extra burden to hyperresponsive lungs.

Topics: Air Pollutants; Air Pollution; Animals; Bronchial Hyperreactivity; Coal Ash; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Function Tests

The Toll-like receptor (TLR) 5 agonist flagellin is associated with immunomodulatory functions.. We sought to investigate whether Listeria monocytogenes-derived flagellin A (flaA) can modulate ovalbumin (OVA)-specific T-cell responses and prevent OVA-induced intestinal allergy.. Bone marrow-derived myeloid dendritic cells from BALB/c, C57BL/6, or TLR signaling-deficient (MyD88(-/-)) mice were stimulated with rOVA, rflaA, rflaA plus rOVA, or a recombinant fusion protein consisting of rflaA and rOVA (rflaA:OVA). The immunomodulating properties of rflaA plus rOVA and rflaA:OVA were investigated by means of DC-T-cell coculture with CD4(+) T cells from OVA-T-cell receptor transgenic or OVA/alum-immunized mice. rflaA:OVA was applied as a prophylactic and therapeutic vaccine in a murine model of intestinal allergy.. rflaA:OVA induced upregulation of TLR5 and dose-dependent IL-6 and IL-10 secretion by myeloid dendritic cells. IL-10 contributed to repressing IL-4 and IFN-γ secretion by OVA-T-cell receptor transgenic CD4(+) T cells. Moreover, rflaA:OVA suppressed CD4(+) T cells derived from T(H)2-biased mice on OVA/alum immunization. In a murine model of intestinal allergy, prophylactic vaccination with rflaA:OVA reduced T-cell activation. Protection from intestinal allergy included suppression of OVA-specific IgE while inducing OVA-specific IgG(2a). Equimolar amounts of rflaA or rOVA provided alone or as a mixture did not have comparable effects. Moreover, therapeutic vaccination was shown to reduce allergic symptoms and T-cell activation in the spleen.. The rflaA:OVA fusion protein showed strong TLR-mediated immunomodulating capacities probably attributed by the proximity of adjuvant and allergen, leading to the prevention of intestinal allergy in a murine disease model. Therefore recombinant flaA:allergen fusion proteins are promising vaccine candidates for intervention in patients with IgE-mediated allergy.

Topics: Adjuvants, Immunologic; Allergens; Animals; Flagellin; Hypersensitivity; Immunologic Factors; Intestines; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Ovalbumin; Recombinant Fusion Proteins; Th2 Cells; Vaccines

Reports have recently suggested that eosinophils have the potential to modulate allergen-dependent pulmonary immune responses. The studies presented expand these reports demonstrating in the mouse that eosinophils are required for the allergen-dependent Th2 pulmonary immune responses mediated by dendritic cells (DCs) and T lymphocytes. Specifically, the recruitment of peripheral eosinophils to the pulmonary lymphatic compartment(s) was required for the accumulation of myeloid DCs in draining lymph nodes and, in turn, Ag-specific T effector cell production. These effects on DCs and Ag-specific T cells did not require MHC class II expression on eosinophils, suggesting that these granulocytes have an accessory role as opposed to direct T cell stimulation. The data also showed that eosinophils uniquely suppress the DC-mediated production of Th17 and, to smaller degree, Th1 responses. The cumulative effect of these eosinophil-dependent immune mechanisms is to promote the Th2 polarization characteristic of the pulmonary microenvironment after allergen challenge.

Topics: Adoptive Transfer; Allergens; Animals; Asthma; Cell Separation; Chemotaxis, Leukocyte; Dendritic Cells; Eosinophils; Flow Cytometry; Hypersensitivity; Immunity, Mucosal; Lung; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Th2 Cells

Several studies have demonstrated that herbal extracts possess various biological effects including anti-inflammatory and anti-cancer activities. The present study was aimed to investigate the protective effects of the Astragalus gypsicolus (AG) hydroalcoholic extract in early allergic sensitized mice induced by ovalbumin. Phytochemical assay was used to recognize the main active constituents in the AG hydroalcoholic extract. Mice were immunized with subcutaneous injection of ovalbumin and aluminum hydroxide. Efficiency of sensitization was assessed by serum IgE levels and eosinophil count. After sensitization, two doses of extract (250 mg/kg and 500 mg/kg) were injected intrapritoneally. On day 14, mice were challenged with intrapritoneal injection of ovalbumin. IL-4 and IFNγ levels in broncoalveolar lavage fluid, which had been collected on day 15, were assessed by Enzyme-Linked Immunosorbent Assay (ELISA) kit. Our results indicate two main active constituents including flavonoids and terpenoids are present in the AG hydroalcoholic extract. Intrapritoneal injection of the AG hydroalcoholic extract was able to decrease IL-4 and increase IFNγ. It seems the AG hydroalcoholic extract has the potential to modulate the balance of Th1/Th2 cytokines in allergy.

Topics: Animals; Astragalus Plant; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hypersensitivity; Immunologic Factors; Interferon-gamma; Interleukin-4; Male; Mice; Ovalbumin; Plant Extracts

It is well documented that exposure to ultraviolet (UV) radiation in sunlight before immunization suppresses systemic as well as local immune responses. We have previously shown that administrating UV irradiation 7 days after immunization also suppresses Th1- and Th2-driven antibody (Ab) via generation of antigen (Ag)-specific CD4(+) regulatory T cells. In this study, we specifically show that IL-10, which is produced by CD4(+) regulatory T cells generated in mice that received UV irradiation after immunization, mediates the suppression of Ab responses by inhibiting Th cell activation. In addition, IL-10 produced upon Ag-specific activation by UV-induced regulatory T cells also mediates bystander suppression. Furthermore, because UV irradiation after immunization effectively dampens both Th1 and Th2 immune responses, we further demonstrated that mice receiving UV irradiation after allergen sensitization had reduced Th2-driven airway inflammation and airway hyperreactivity (AHR). These results suggest that UV irradiation in pre-sensitized individuals induces Ag-specific IL-10 producing regulatory T cells representing type 1 regulatory T cells that suppress Th2 immunity and may have therapeutic potential for asthmatic patients.

Topics: Animals; Down-Regulation; Female; Hypersensitivity; Immunity, Active; Immunization; Immunosuppression Therapy; Interleukin-10; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells; Ultraviolet Rays

Violacein was isolated from Chromobacterium violaceum, a soil Gram negative bacterium collected from the forest water body soil sample of Kolli Hills; Tamil Nadu, India. In the present study the immunomodulatory, analgesic and antipyretic activities of violacein were investigated in wistar rats and mice. Analgesic effect was evaluated by acetic acid- induced writhing, formalin induced paw licking and hotplate tests. Immunomodulatory effect was investigated by using ovalbumin- induced active paw anaphylaxis and sheep red blood cells (SRBC)-induced DTH tests. Antipyretic activity was evaluated by yeast- induced hyperpyrexia in rats. The anti- oedema effect was compared with indomethacin. Violacein inhibited 42.9% of ovalbumin- induced edema. Further we found that violacein (40mg/kg b.w.) reduced the edema induced by sheep red blood cells. Violacein also produced significant (p<0.05) analgesic activity in acetic acid induced writhing response, formalin induced paw licking response and hot plate analysis. Treatment with violacein showed a significant (p<0.05) dose-dependent reduction in pyrexia in rats. The results suggest that violacein possesses potent immunomodulatory, analgesic and antipyretic activities.

Topics: Acetic Acid; Analgesics; Anaphylaxis; Animals; Behavior, Animal; Chromobacterium; Dose-Response Relationship, Drug; Edema; Erythrocytes; Female; Fever; Formaldehyde; Hot Temperature; Hypersensitivity; Hypersensitivity, Delayed; Immunologic Factors; India; Indoles; Indomethacin; Male; Mice; Ovalbumin; Pain; Rats; Rats, Wistar; Sheep; Yeasts

Mycobacterium bovis Bacillus Calmette-Guerin (BCG) has been shown to down-regulate experimental allergic asthma, a finding that reinforced the hygiene hypothesis. We have previously found that recombinant BCG (rBCG) strain that express the genetically detoxified S1 subunit of pertussis toxin (rBCG-S1PT) exerts an adjuvant effect that enhances Th1 responses against BCG proteins. Here we investigated the effect of this rBCG-S1PT on the classical ovalbumin-induced mouse model of allergic lung disease. We found that rBCG-S1PT was more effective than wild-type BCG in preventing Th2-mediated allergic immune responses. The inhibition of allergic lung disease was not associated with increased concentration of suppressive cytokines or with an increased number of pulmonary regulatory T cells but was positively correlated with the increase in IFN-gamma-producing T cells and T-bet expression in the lung. In addition, an IL-12-dependent mechanism appeared to be important to the inhibition of lung allergic disease. The inhibition of allergic inflammation was found to be restricted to the lung because when allergen challenge was given by the intraperitoneal route, rBCG-S1PT administration failed to inhibit peritoneal allergic inflammation and type 2 cytokine production. Our work offers a nonclassical interpretation for the hygiene hypothesis indicating that attenuation of lung allergy by rBCG could be due to the enhancement of local lung Th1 immunity induced by rBCG-S1PT. Moreover, it highlights the possible use of rBCG strains as multipurpose immunomodulators by inducing specific immunity against microbial products while protecting against allergic asthma.

Topics: Animals; Cytokines; Eosinophils; Female; Hypersensitivity; Interferon-gamma; Interleukin-12; Mice; Mice, Inbred BALB C; Mice, Knockout; Mycobacterium bovis; Ovalbumin; Recombinant Proteins; Th1 Cells; Th2 Cells

Studies in humans and rodents have indicated a causative role for CD8(+) T cells in IgE-mediated allergic inflammation, but their function is still controversial.. To analyze the role of allergen-specific CD8(+) T cells during the development of allergic airway inflammation in two parallel but diverging outcome models.. We used H2-Kb SIINFEKL (OVA(257-264)) multimers to analyze induction, natural distribution, and phenotype of allergen-specific CD8(+) T cells in a murine C57BL/6 model of ovalbumin (OVA)-induced allergic airway inflammation using low-dose or high-dose OVA sensitization.. The low-dose protocol was characterized by a significant induction of total and OVA-specific IgE, eosinophilic airway inflammation, IL-4 levels in bronchoalveolar lavage fluid. And significant alterations in lung function. The high dose protocol was characterized by a significant reduction of the allergic phenotype. Using OVA(257-264) H2-Kb multimers, we observed lung and airway infiltrating OVA-specific CD8(+) T cells showing an effector/effector-memory phenotype. The high-dose protocol caused significantly higher infiltration of allergen-specific CD8(+) cells to the airways and enhanced their cytotoxicity. Adoptive transfer with CD8(+) T cells from transgenic OT-I mice to TAP1(-/-) or wild-type mice showed their migration to the lungs and TAP1-dependent proliferation after OVA-aerosol exposure. TAP1(-/-) mice defective in CD8(+) T cells showed exacerbated symptoms in the low-dose sensitization model.. Allergen-specific CD8(+) T cells seem to protect from allergic inflammation in the lungs. Their number, which is dependent on the sensitization dose, appears to be a critical predictor for the severity of the allergic phenotype.

Topics: Allergens; Animals; ATP Binding Cassette Transporter, Subfamily B, Member 2; ATP-Binding Cassette Transporters; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; CD8-Positive T-Lymphocytes; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Immunologic Memory; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Peptide Fragments

Asthma is characterized by airway inflammation, mucus overproduction, airway hyperreactivity, and peribronchial fibrosis. Intelectin has been shown to be increased in airway epithelium of asthmatics. However, the role of intelectin in the pathogenesis of asthma is unknown. Airway epithelial cells can secrete chemokines such as monocyte chemotactic protein (MCP)-1 and -3 that play crucial roles in asthmatic airway inflammation. We hypothesized that intelectin plays a role in allergic airway inflammation by regulating chemokine expression. In a mouse allergic asthma model, we found that mRNA expression of intelectin-2 as well as MCP-1 and -3 in mouse lung was increased very early (within 2 h) after allergen challenge. Expression of intelectin protein was localized to mucous cells in airway epithelium. Treatment of MLE12 mouse lung epithelial cells with interleukin IL-13, a critical mediator of allergic airway disease, induced expression of intelectin-1 and -2 as well as MCP-1 and -3. When IL-13-induced intelectin-1 and -2 expression was inhibited by RNA interference, IL-13-induced extracellular signal-regulated kinase 1/2 phosphorylation and MCP-1 and -3 production by MLE12 cells was inhibited. Furthermore, inhibition of intelectin expression by airway transfection with shRNA targeting intelectin-1 and -2 attenuated allergen-induced airway inflammation. We conclude that intelectin, a molecule expressed by airway epithelial cells and upregulated in asthma, is required for IL-13-induced MCP-1 and -3 production in mouse lung epithelial cells and contributes to allergic airway inflammation.

Topics: Allergens; Animals; Chemokine CCL2; Chemokine CCL7; Enzyme Activation; Epithelial Cells; Hypersensitivity; Interleukin-13; Kinetics; Lectins; Lung; Mice; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase 6; Mucous Membrane; Ovalbumin; Pneumonia; Protein Transport; Up-Regulation

Several differences have been described between neonatal and adult immune responses. The predisposition in early life to Th2-type response or tolerance makes it a susceptible period for infections and allergic sensitization.. The aim of this work was to evaluate the effects of CpG-containing oligodeoxynucleotides on neonatal and adult immunization with ovalbumin and Blomia tropicalis extract and compare the CpG effects on B and T cells of neonatal and adult mice.. Mice that received CpG showed reduced immunoglobulin E (IgE) antibody production in both neonatal and adult periods, in parallel to increased IgG2a antibody levels. We observed that spleen cells of mice that received CpG in early life produced increased amounts of interferon-gamma upon anti-CD3 stimulation. Negative regulation of IgE response was more pronounced in adult than neonate mice; further, CpG decreased anaphylactic antiovalbumin IgG1 only in adults. Also, an upregulation of toll-like receptor 9 expression was detected in adult B cells, but not in neonatal, upon CpG stimuli. Neonatal B cells showed enhanced interleukin (IL)-10 expression and decreased IL-6 levels than adult B cells in response to CpG. When we analyzed in vitro activation of CD4+ T cells, an increased expression of B7 molecules on T cells in neonates was suppressed by CpG.. Altogether, we verified qualitative and quantitative evidences regarding CpG effect on neonatal and adult allergens immunizations, which points to the importance of understanding neonatal immune system to establish immunomodulatory strategies for prevention of allergic diseases.

Topics: Animals; Animals, Newborn; Antigens, Dermatophagoides; B-Lymphocytes; B7-1 Antigen; Cell Extracts; DNA; Female; Hypersensitivity; Immunity, Humoral; Immunization; Immunoglobulin E; Mice; Mice, Inbred Strains; Oligodeoxyribonucleotides; Ovalbumin; Pyroglyphidae; T-Lymphocytes; Toll-Like Receptor 9

Anoectochilus formosanus HAYATA, a Chinese herb, is a valued folk medicine for fever, pain, and diseases of the lung and liver. Allergic asthma is characterized by increased serum IgE level and inflammation of the airways with high levels of interleukin (IL)-4 and IL-5 in bronchoalveolar lavage fluids (BALF). Constriction of airway smooth muscle and development of airway hyperresponsiveness (AHR) are the most important symptoms of allergic asthma. In our previous study, a standardized aqueous extract of A. formosanus (SAEAF) was used to modulate innate immunity of normal mice. In this study, airway inflammatory infiltrations, including T cell differentiation, cytokine modulation, allergic antibodies estimation, pulmonary pathology, and enhanced pause (Penh) of AHR were used to evaluate SAEAF treatment of an ovalbumin (OVA)-inhaled airway allergic murine model. The resulting cytokine profiles demonstrated that SAEAF can significantly reduce Th2 polarization after administration of SAEAF in OVA inhalation. These results also suggest that SAEAF modulates cytokine secretion in allergic asthma. Modulated natural T regulatory cells (CD25+/CD4+, Treg) were also shown to increase immuno-suppression in the allergic lung inflammation and further down-regulate airway inflammatory infiltration in eosinophils and macrophages. Finally, decreased airway anti-OVA IgE secretion and reduced AHR were observed. Our results indicate that the administration of SAEAF can modulate cytokines and T cell subpopulation by regulating inflammatory cell infiltration and modulating the allergic response.

Topics: Animals; Anti-Inflammatory Agents; Antigens; Asthma; Bronchial Hyperreactivity; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Eosinophils; Hypersensitivity; Immunoglobulin E; Immunosuppressive Agents; Lung; Macrophages; Male; Mice; Mice, Inbred BALB C; Orchidaceae; Ovalbumin; Phytotherapy; Reference Values; T-Lymphocytes, Regulatory; Th2 Cells

The herbal Isatis tinctoria extract (ITE) inhibits the inducible isoform of cyclooxygenase (COX-2) as well as lipoxygenase (5-LOX) and therefore possesses anti-inflammatory properties. The extract might also be useful in allergic airway diseases which are characterized by chronic inflammation.. ITE obtained from leaves by supercritical carbon dioxide extraction was investigated in ovalbumin (OVA) immunised BALB/c mice given intranasally together with antigen challenge in the murine model of allergic airway disease (asthma) with the analysis of the inflammatory and immune parameters in the lung.. ITE given with the antigen challenge inhibited in a dose related manner the allergic response. ITE diminished airway hyperresponsiveness (AHR) and eosinophil recruitment into the bronchoalveolar lavage (BAL) fluid upon allergen challenge, but had no effect in the saline control mice. Eosinophil recruitment was further assessed in the lung by eosinophil peroxidase (EPO) activity at a dose of 30 microg ITE per mouse. Microscopic investigations revealed less inflammation, eosinophil recruitment and mucus hyperproduction in the lung in a dose related manner. Diminution of AHR and inflammation was associated with reduced IL-4, IL-5, and RANTES production in the BAL fluid at the 30 microg ITE dose, while OVA specific IgE and eotaxin serum levels remained unchanged.. ITE, which has been reported inhibiting COX-2 and 5-LOX, reduced allergic airway inflammation and AHR by inhibiting the production of the Th2 cytokines IL-4 and IL-5, and RANTES.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Eosinophil Peroxidase; Eosinophils; Hypersensitivity; Immunoglobulin E; Inflammation; Inflammation Mediators; Isatis; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Phytotherapy; Plant Extracts; Plant Leaves

Obesity is associated with deterioration in asthma outcomes. Although airways eosinophil accumulation is characteristic of lung allergic diseases, little is known about the influence of obesity on the allergic eosinophil trafficking from bone marrow to lung tissues, and recruitment to airways lumen. Here, we have assessed the effects of diet-induced obesity on allergic eosinophilic inflammation in mice, examining eosinophil trafficking from bone marrow to airways, and production of T(H)1/T(H)2 cytokines.. C57BL/6 mice fed for 10 weeks with standard chow or high-fat diet were sensitized and challenged with ovalbumin. At 24-96 h post-ovalbumin challenge, bronchoalveolar lavage (BAL) fluid, lung tissue and bone marrow were examined.. The high-fat-fed mice exhibited increased body weight and epididymal fat, glucose intolerance and alterations in lipid profile compared with the lean mice. Obesity markedly elevated serum leptin and lowered adiponectin levels. Ovalbumin challenge in obese mice promoted a markedly higher eosinophil accumulation in bone marrow and connective tissue surrounding the bronchial and bronchiolar segments. Eosinophil number in BAL fluid of obese mice was lower at 24 and 48 h. Levels of interleukin (IL)-5, eotaxin, tumour necrosis factor-alpha and IL-10 in BAL fluid of obese mice were significantly higher than in lean mice.. Diet-induced obesity enhanced eosinophil trafficking from bone marrow to lung tissues, and delayed their transit through the airway epithelium into the airway lumen. Consequently, eosinophils remain longer in lung peribronchiolar segments due to overproduction of T(H)1/T(H)2 cytokines and chemokines.

Topics: Animals; Asthma; Bone Marrow; Bronchi; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Cytokines; Eosinophilia; Eosinophils; Hypersensitivity; Inflammation; Interleukin-10; Interleukin-5; Lung; Lung Diseases; Male; Mice; Mice, Inbred C57BL; Obesity; Ovalbumin; Pneumonia; Tumor Necrosis Factor-alpha

Asthma is characterized by difficulty in breathing because of the constriction of the smooth muscles of the bronchi, as a result of inflammation. In the present study, we focus on the protective effects of allantoin against ovalbumin (OVA)-induced lung inflammation in a murine allergic model, and assess cytokine release, eosinophilia, and mucus hypersecretion. Allantoin treatment led to significant reduction in the levels of Ig(immunoglobulin)E and T-helper-2-type cytokines, such as IL(interleukin)-4 and IL-5, in bronchoalveolar lavage (BAL) fluid. Airway inflammatory-cell infiltration was remarkably alleviated in allantoin-treated asthma groups, compared with the control group. Moreover, allantoin-treated asthma groups exhibited a marked decrease in cytokine mRNA expression in lung tissues, compared with the control group. The effectiveness of allantoin was similar to that of montelukast, used as a positive control. These results support the utility of allantoin as a protective agent against asthma.

Topics: Alanine Transaminase; Allantoin; Animals; Aspartate Aminotransferases; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Female; Goblet Cells; Hypersensitivity; Image Processing, Computer-Assisted; Immunoglobulin E; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Pneumonia; Reverse Transcriptase Polymerase Chain Reaction; Th2 Cells

Although whole-parasite vaccine strategies for malaria infection have regained attention, their immunological mechanisms of action remain unclear. We find that immunization of mice with a crude blood stage extract of the malaria parasite Plasmodium falciparum elicits parasite antigen-specific immune responses via Toll-like receptor (TLR) 9 and that the malarial heme-detoxification byproduct, hemozoin (HZ), but not malarial DNA, produces a potent adjuvant effect. Malarial and synthetic (s)HZ bound TLR9 directly to induce conformational changes in the receptor. The adjuvant effect of sHZ depended on its method of synthesis and particle size. Although natural HZ acts as a TLR9 ligand, the adjuvant effects of synthetic HZ are independent of TLR9 or the NLRP3-inflammasome but are dependent on MyD88. The adjuvant function of sHZ was further validated in a canine antiallergen vaccine model. Thus, HZ can influence adaptive immune responses to malaria infection and may have therapeutic value in vaccine adjuvant development.

Topics: Adjuvants, Immunologic; Animals; Antibodies, Protozoan; Carrier Proteins; Hemeproteins; Hypersensitivity; Malaria Vaccines; Malaria, Falciparum; Mice; Mice, Inbred C57BL; Mice, Knockout; Myeloid Differentiation Factor 88; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Plasmodium falciparum; Protein Binding; Protein Conformation; Toll-Like Receptor 9

We investigated the anti-allergic effect of a new strain (Pediococcus pentosaceus Sn26, the Sn26 strain) among 59 strains isolated from Japanese fermented vegetable pickles, the Sunki pickle. The Sn26 strain increased Th1 type cytokine (IL-12 and IFN-gamma) production of Peyer's patch (PP) cells in BALB/c mice, improved the Th1/Th2 balance, and inhibited IgE production of splenocytes of ovalbumin (OVA)-induced allergic diarrheic mice. Next we demonstrated, by neutralizing IL-12 and IFN-gamma, that the Sn26 strain first induced IL-12, that IL-12 induced IFN-gamma, and that decreases in IL-4 and IgE production followed. Furthermore, oral administration of the Sn26 strain decreased serum OVA-specific IgE levels and ameliorated the appearance of diarrhea in OVA-induced allergic diarrheic mice. Based on these results, it was assumed that oral administration of the Sn26 strain ameliorated type-1 allergies through improvement of the Th1/Th2 balance and decreases in IgE production.

Topics: Animals; Diarrhea; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Pediococcus

Interleukin-5 (IL-5) is a cytokine which is essential for the maturation of eosinophils in bone marrow and for their release into the blood. Eotaxin is a CC type chemokine implicated in the recruitment of eosinophils in a variety of inflammatory disorders. Since eosinophil-activity is governed by these two pathways, we targeted both IL-5 and eotaxin by active vaccination to block eosinophilia. We produced two vaccines by chemically cross-linking IL-5 or eotaxin to a virus-like particle (VLP) derived from the bacteriophage Qbeta, yielding highly repetitive arrays of these cytokines on the VLP surface. Both vaccines overcame self-tolerance and induced high antibody titers against the corresponding self-molecules in mice. Immunization with either of the two vaccines reduced eosinophilic inflammation of the lung in an ovalbumin (OVA) based mouse model of allergic airway inflammation. Animals immunized with the two vaccines at the same time developed high antibody titers against both cytokines and also reduced eosinophil-infiltration of the lung. These data demonstrate that targeting either IL-5 or eotaxin may lower eosinophilia. Simultaneous immunization against IL-5 and eotaxin demonstrates that such a therapeutic approach may be used to treat complex disorders in which multiple mediators are involved.

Topics: Allergens; Allolevivirus; Animals; Autoantibodies; Chemokine CCL11; Eosinophilia; Female; Hypersensitivity; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Tract Diseases; Vaccination; Vaccines, Virosome; Viral Proteins

Allergy has been an increasing problem in several parts of the world. Prenatal exposure to allergen and microbial components may affect the development of allergies in childhood, as indicated by epidemiological and experimental studies. We investigated the capacity for allergic sensitisation in offspring after induction of a Th1- or a Th2-polarised immune response to the same allergen in mothers during pregnancy.. During pregnancy, mice were immunised with ovalbumin (OVA) given with either one of the Th2-adjuvants pertussis toxin (PT) or Al(OH)3 (aluminium hydroxide), or with the Th1 adjuvant CpG. Offspring were immunised with OVA in Al(OH)3 as young adults. Serum and supernatants from ex vivo stimulated or non-stimulated spleen cells from mothers and offspring were analysed for OVA-specific antibodies and cytokines, respectively. Mothers immunised with OVA together with either Al(OH)3 or PT had increased levels of OVA-specific IgE and IgG1 compared to naive mothers, whereas mothers immunised with OVA together with CpG had increased levels of OVA-specific IgG2a compared to naive mothers. In general the highest levels of IL-5, IL-10, and IFNgamma were observed in spleen cells from mothers immunised with PT and OVA. Upon immunisation, offspring from mothers immunised with OVA and either PT or Al(OH)3 showed reduced levels of OVA-specific IgE and IgG1 and increased levels of OVA-specific IgG2a antibodies compared to offspring from naive mothers. Maternal immunisation with CpG and OVA did not affect antibody responses in offspring.. Allergic sensitisation in the offspring was affected by the type of adjuvant used for immunisation of the mothers with the same allergen. Th2 polarisation of the immune response in the mothers was found to give reduced IgE levels upon sensitisation of the offspring, whereas no reduction was achieved with Th1 polarisation in the mothers.

Topics: Adjuvants, Immunologic; Allergens; Aluminum Hydroxide; Animals; Cytokines; DNA; Female; Gene Expression Regulation, Developmental; Hypersensitivity; Immunity, Maternally-Acquired; Immunization; Immunoglobulin E; Male; Mice; Oligodeoxyribonucleotides; Ovalbumin; Pertussis Toxin; Pregnancy; Prenatal Exposure Delayed Effects; Th1 Cells; Th2 Cells

A proinflammatory role of T helper (Th)17 cells, producing IL-22 and IL-17A, has been favored although there is evidence for negative immune regulation by IL-17A. Here we show that IL-22 was produced during an allergic response in lungs of mice, immunized and challenged with ovalbumin (OVA), and that IL-22 neutralization further augmented the eosinophil recruitment to the lung. In a second allergy model, transfer of OVA-pulsed dendritic cells (DC) into naive mice conveyed eosinophil recruitment in response to subsequent inhaled OVA challenge, while DC preincubation with recombinant IL-22 abolished this response. Similarly, DC preincubation with IL-17A abolished DC-driven eosinophil recruitment, showing that both Th17 cytokines IL-22 and IL-17A mediate negative regulation of allergy by acting on DCs. Therefore, IL-22 inhibits DC functions and attenuates an allergic response.

Topics: Animals; Cell Movement; Dendritic Cells; Eosinophils; Hypersensitivity; Interleukin-22; Interleukin-4; Interleukins; Lymph Nodes; Mediastinum; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction; Th1 Cells; Th2 Cells

Allergic asthma is an inflammatory lung disease driven by Th2. We have shown that both Th1 and Th2 sensitization to inhaled OVA depend on the presence and concentration of LPS, where high concentrations (LPS(hi)) induce Th1 and low concentrations (LPS(lo)), Th2. Stromal cells (SCs), such as airway SCs, exacerbate established airway disease; however, little is known about their role early during sensitization. In this study, using bone marrow chimeric mice to restrict TLR4 signaling to either the SC compartment (SC(+)HPC(-)) or the hematopoietic cell (HPC) compartment (SC(-)HPC(+)), we report that HPC TLR4 is necessary and sufficient for Th1 sensitization to OVA-LPS(hi), whereas TLR4 in both compartments is required for Th2 sensitization to OVA-LPS(lo). Surprisingly, although SC(+)HPC(-) mice were unable to generate a Th1 response to OVA-LPS(hi), they instead mounted a robust Th2 response, indicating that in the presence of higher concentrations of LPS, SC TLR4 is sufficient for Th2 sensitization. We show that the SC TLR4 response to LPS leads to induction of Th2-inducing dendritic cells that upregulate Notch ligand Jagged-1 but not Delta-4. Furthermore, airway SCs upregulate thymic stromal lymphopoietin in response to exposure to both OVA-LPS(lo) and OVA-LPS(hi). These studies demonstrate that SC TLR4 signaling is critically involved in Th2 but not Th1 sensitization to inhaled Ag.

Topics: Administration, Inhalation; Animals; Antigens; Asthma; Cell Differentiation; Chemotaxis, Leukocyte; Coculture Techniques; Dendritic Cells; Female; Gene Expression; Hypersensitivity; Lung; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Stromal Cells; Th2 Cells; Toll-Like Receptor 4; Transplantation Chimera

The formylpeptide receptor-like 1, now officially termed FPR2, in human and its mouse homolog mFPR2 mediate leukocyte migration in response to agonists associated with inflammation and immune responses. To clarify the in vivo role of the receptor, we generated mice deficient in mFPR2. mFPR2(-/-) mice showed markedly reduced severity in OVA/alum-induced allergic airway inflammation. This was associated with diminished recruitment of CD11c(+) dendritic cells into the airway mucosa and secondary lymphoid organs, as well as reduced production of Type 2 cytokines and Igs. We also found that the bronchoalveolar lavage fluid from wild type mice with airway inflammation contained mFPR2 agonist activity. This study reveals a critical role for mFPR2 in the progression of allergic airway inflammation and immune responses.

Topics: Allergens; Alum Compounds; Animals; Bronchoalveolar Lavage Fluid; Cell Separation; Chemotaxis, Leukocyte; Cytokines; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fluorescent Antibody Technique; Hypersensitivity; Immunohistochemistry; Inflammation; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Formyl Peptide; Respiratory Hypersensitivity; Reverse Transcriptase Polymerase Chain Reaction

As adjuvant during sensitization may cause unspecific immune reactions, the aim of the present study was to define the role of cyclooxygenase (COX) activity on airway inflammation and airway hyperresponsiveness (AHR) in an adjuvant-free allergic mouse model. Administration of diclofenac and indomethacin (non-selective COX inhibitors), FR122047 (COX-1 inhibitor) and lumiracoxib (selective COX-2 inhibitor) enhanced AHR. Only diclofenac and lumiracoxib reduced the inflammatory cell content of bronchoalveolar lavage (BAL). Moreover, levels of prostaglandins in BAL were reduced by indomethacin and FR122047 but were unaffected by lumiracoxib. However, compared with antigen controls, none of the COX inhibitors displayed major effects on the production of cytokines, smooth muscle mass, number of goblet cells and eosinophils, or collagen deposition in the airways. These data in mice sensitized without adjuvant support the fact that COX products have a general bronchoprotective role in allergic airway inflammation. Furthermore, the data suggest that COX-1 activity predominantly generates prostanoids in BAL, whereas COX-2 activity is associated with the accumulation of inflammatory cells in BAL. This study further supports that AHR on the one hand, and the inflammatory response and generation of prostanoids on the other, are dissociated and, at least in part, uncoupled events.

Topics: Adjuvants, Immunologic; Animals; Bronchoalveolar Lavage; Cyclooxygenase 1; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Cysteine; Cytokines; Female; Hypersensitivity; Immunization; Inflammation; Leukotrienes; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Prostaglandins; Respiratory System

Preconception allergen immunization prevents neonatal allergen sensitization in mice by a complex interaction between regulatory cells/factors and antibodies. The present study assessed the influence of maternal immunization with ovalbumin (OVA) on the immune response of 3 day-old and 3 week-old offspring immunized or non-immunized with OVA and evaluated the effect of IgG treatment during fetal development or neonatal period.. Maternal immunization with OVA showed increased levels of Fc gamma RIIb expression in splenic B cells of neonates, which were maintained for up to 3 weeks and not affected by additional postnatal OVA immunization. Maternal immunization also exerted a down-modulatory effect on both IL-4 and IFN-gamma-secreting T cells and IL-4 and IL-12- secreting B cells. Furthermore, immunized neonates from immunized mothers showed a marked inhibition of antigen-specific IgE Ab production and lowered Th2/Th1 cytokine levels, whereas displaying enhanced Fc gamma RIIb expression on B cells. These offspring also showed reduced antigen-specific proliferative response and lowered B cell responsiveness. Moreover, in vitro evaluation revealed an impairment of B cell activation upon engagement of B cell antigen receptor by IgG from OVA-immunized mice. Finally, in vivo IgG transference during pregnancy or breastfeeding revealed that maternal Ab transference was able to increase regulatory cytokines, such as IL-10, in the prenatal stage; yet only the postnatal treatment prevented neonatal sensitization. None of the IgG treatments induced immunological changes in the offspring, as it was observed for those from OVA-immunized mothers.. Maternal immunization upregulates the inhibitory Fc gamma RIIb expression on offspring B cells, avoiding skewed Th2 response and development of allergy. These findings contribute to the advancement of prophylactic strategies to prevent allergic diseases in early life.

Topics: Allergens; Animals; Animals, Newborn; B-Lymphocytes; CD40 Antigens; Cytokines; Female; Fetal Development; Humans; Hypersensitivity; Immunity, Maternally-Acquired; Immunization; Immunoglobulin E; Infant, Newborn; Infant, Newborn, Diseases; Lymphocyte Activation; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Receptors, IgE; Receptors, IgG; T-Lymphocytes; Up-Regulation

Induction of RNA interference with small interfering RNA (siRNA) has demonstrated therapeutic potential through the knockdown of target genes. We have previously reported that systemic administration of CD40 siRNA is capable of attenuating allergic symptoms but in an allergen-nonspecific fashion. However, siRNA-based allergen-specific therapy for allergy has not been developed.. We attempted to develop a new allergen-specific therapy for allergy using CD40-silenced and allergen-pulsed dendritic cells (DCs).. Bone marrow-derived DCs were silenced with CD40 siRNA and pulsed with ovalbumin (OVA). Mice had allergy after intraperitoneal sensitization with OVA and keyhole limpet hemocyanin, followed by intranasal challenge with the same allergens. The mice were treated with CD40-silenced and OVA-pulsed DCs (CD40-silenced OVA DCs) either before allergic sensitization or after establishing allergic rhinitis.. Mice receiving CD40-silenced OVA DCs either before or after the establishment of allergic rhinitis showed remarkable reductions in allergic symptoms caused by OVA challenge, as well as anti-OVA IgE levels in sera. Additionally, CD40-silenced OVA DCs suppressed eosinophil infiltration at the nasal septum, OVA-specific T-cell responses, T-cell production of IL-4 and IL-5 after stimulation with OVA, and CD4(+)CD25(-) effector T-cell responses. Furthermore, CD40-silenced OVA DCs facilitated the generation of CD4(+)CD25(+) forkhead box protein 3-positive OVA-specific regulatory T cells, which inhibit allergic responses in vivo. However, CD40-silenced OVA DCs suppressed only OVA-specific allergy but did not inhibit keyhole limpet hemocyanin-induced allergy, suggesting that CD40-silenced OVA DCs induce allergen-specific tolerance.. This study is the first to demonstrate a novel allergen-specific therapy for allergy through DC-mediated immune modulation after gene silencing of CD40.

Topics: Animals; CD40 Antigens; Cell Separation; Dendritic Cells; Flow Cytometry; Hypersensitivity; Immunotherapy; Mice; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA, Small Interfering; T-Lymphocyte Subsets; T-Lymphocytes

In recent in vitro reports, the IC(50) value of ayanin (quercetin-3,7,4'-O-trimethylether) was 2.2microM for inhibiting interleukin (IL)-4 production from purified basophils, and its therapeutic ratio was >19. Therefore, we were interested in investigating the effects on ovalbumin induced airway hyperresponsiveness in vivo, and to clarify its potential for treating asthma. Ayanin (30-100micromol/kg, orally (p.o.)) dose-dependently and significantly attenuated the enhanced pause (P(enh)) value induced by methacholine in sensitized and challenged mice. It also significantly suppressed the increases in total inflammatory cells, macrophages, lymphocytes, neutrophils, and eosinophils, and levels of cytokines, including IL-2, IL-4, IL-5, and tumor necrosis factor (TNF)-alpha in bronchoalveolar lavage fluid of these mice. However, at 100micromol/kg, it significantly enhanced the level of interferon (IFN)-gamma. In addition, ayanin (30-100micromol/kg, p.o.) dose-dependently and significantly suppressed total and OVA-specific immunoglobulin (Ig)E levels in the serum and bronchoalveolar lavage fluid, and enhanced the IgG(2a) level in serum of these mice. In the present results, ayanin did not affect xylazine/ketamine-induced anesthesia, suggesting that ayanin has few or no adverse effects, such as nausea, vomiting, and gastric hypersecretion. In conclusion, the above results suggest that ayanin may have the potential for use in treating allergic asthma.

Topics: Anesthesia; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Flavonoids; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Ketamine; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Respiratory System; Xylazine

Tamoxifen (TX) represents the prototype selective oestrogen receptor modulator. In addition to its use in breast cancer, TX possesses immunomodulatory functions and displays beneficial effects in models of systemic lupus erythematosus. We hypothesized that TX might inhibit type I allergic reactions, which are also characterized by deviations in humoral immunity.. To evaluate the effects of TX on the allergic immune response in appropriate mouse models.. Balb/c mice were sensitized with ovalbumin (OVA)-alum by the intraperitoneal route, and humoral parameters, T cell cytokine patterns and OVA-induced ear swelling responses were determined in a preventive (start of TX treatment before sensitization) and a therapeutic setting (start after sensitization), respectively. In addition, the impact of TX on clinical signs, epidermal thickness and leucocyte infiltration of the skin was investigated in a model of allergen-induced dermatitis.. Preventive TX treatment interfered with all aspects of the allergic immune response, leading to a reduction of allergen-specific Ig levels (IgE, IgG1 and IgG2a), a skewing effect in the T cell compartment with the inhibition of IL-4 and an abrogation of ear swelling responses. Interestingly, a therapeutic TX administration was also effective in reducing Ig levels and ear swelling responses. The vigorous systemic effects were additionally mirrored by local changes in allergen-dependent dermatitis with reduced clinical symptoms, diminished epidermal thickness and decreased CD4+ and CD8+ cell infiltrates.. TX inhibits allergic responses when given preventively and also therapeutically, and improves allergen-induced dermatitis. Because of its effectiveness, TX could bear significant therapeutic potential for the treatment of allergies.

Topics: Allergens; Animals; Anti-Allergic Agents; Cytokines; Dermatitis, Contact; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes; Tamoxifen

Schistosoma mansoni infection modulates the immunity to unrelated antigens in the host. In this study, we have investigated the effect of pregnancy and nursing from schistosomotic mother mice on the immune response to ovalbumin (OA), in adult offspring. Then, newborn mice were divided into four groups: animals born from infected mothers (BIM) suckled by non-infected mothers; animals from non-infected mothers suckled by infected mothers (SIM); and two other groups that were mice born and suckled in infected mothers (BSIM) or non-infected (control) mothers. The adult offspring were immunized with OA plus adjuvant. We compared the OA-specific hypersensitivity reactions (HR), antibodies levels (IgG, IgG2a) and the cytokine production in splenocyte cultures. Remarkable interleukin (IL)-10 synthesis was observed in mice BIM; while the anti-OA antibodies levels and immediate HR were impaired. IL-10 neutralization recovered this suppression. Differently, in mice SIM and BSIM there was an enhancement in the anti-OA humoral response and high IL-2 production, however low level of the IL-10 was detected in mice BSIM. In conclusion, schistosomotic pregnancy provides an immunosuppressive potential, IL-10 dependent, which was sustained throughout adult life. Regardless, suckling by infected mothers induces great responsiveness to an unrelated antigen and repairs the inhibitory potential acquired during prenatal stage.

Topics: Animals; Antibodies; Cytokines; Female; Hypersensitivity; Immune Tolerance; Immunoglobulin G; Leukocytes, Mononuclear; Mice; Ovalbumin; Pregnancy; Pregnancy Complications, Parasitic; Schistosoma mansoni; Schistosomiasis mansoni; Spleen

Histamine controls the function of dendritic cells (DCs). It appears to be required for the normal development of DCs. It also induces the chemotaxis of immature DCs and promotes the differentiation of CD4(+) T cells into cells with a T helper type 2 (Th2) profile. Moreover, we have recently shown that histamine stimulates both the uptake and the cross-presentation of antigens by DCs, supporting the theory that histamine promotes activation of CD8(+) T cells during the development of allergic pathologies. Here, we investigated whether the course of an allergic response, in a well-defined model of ovalbumin (OVA)-induced allergic airway inflammation, could be modulated by intratracheal injection of OVA-pulsed DCs previously treated with histamine (DCHISs). Compared with control DCs, DCHISs induced: (i) greater recruitment of CD8(+) T cells in the lung, (ii) greater stimulation of the production of interleukin (IL)-5 by lung CD8(+) T cells, and (iii) increased recruitment of CD11c/CD8 double-positive DCs in the lungs of allergic mice. Moreover, mice treated with DCHISs showed increased levels of serum-specific immunoglobulin E (IgE) antibodies directed to OVA, and a higher proportion of eosinophils in bronchoalveolar lavage (BAL) compared with mice treated with OVA-pulsed control DCs. Our results support the notion that histamine, by acting on DCs, increases the severity of allergic processes.

Topics: Animals; CD8-Positive T-Lymphocytes; Cells, Cultured; Dendritic Cells; Female; Histamine; Hypersensitivity; Lung Diseases; Mice; Mice, Inbred BALB C; Ovalbumin

Around 300 million people world-wide suffer from asthma, and the prevalence of allergic diseases has increased. Much effort has been used in the study of mechanisms involved in the immune response observed in asthma to intervene for the treatment of this condition. During inflammation in asthma, Th2 cytokines and eosinophils are essential components of the host immune system. Furthermore, for therapeutic interventions against this disease, IL-10 is an important cytokine because it has a central role in the regulation of inflammatory cascades.. To evaluate the immunomodulatory effect of Lactococcus lactis strains expressing recombinant IL-10 in a mouse model of ovalbumin (OVA)-induced acute airway inflammation.. L. lactis expressing recombinant IL-10 in a cytoplasmic (LL-CYT) or secreted form (LL-SEC) and wild-type (LL-WT) were used. IL-10 production by the recombinant strains was evaluated by ELISA. After an intranasal administration of L. lactis producing recombinant IL-10 and the induction of acute allergic airway inflammation in mice, blood samples were collected to detect IgE anti-OVA, and bronchoalveolar lavage (BAL) was harvested for eosinophil count. Additionally, the lungs were collected for the detection of the eosinophil peroxidase (EPO) activity, measurement of cytokines and chemokines and evaluation of pathology.. Mice that received LL-CYT and LL-SEC strains showed a significant decrease in eosinophils numbers, EPO activity, anti-OVA IgE and IgG1 levels, IL-4 and CCL3 production and pulmonary inflammation and mucus hypersecretion, compared with the asthmatic group. Only the LL-CYT/OVA group showed reduced levels of IL-5, CCL2, CCL5 and CCL11.. Treatment with L. lactis producing recombinant IL-10 used in this study (LL-CYT and LL-SEC) modulated experimental airway inflammation in the mouse model independently of Treg cells. Additionally, the LL-CYT strain was more efficient in the suppression of lung inflammation.

Topics: Administration, Intranasal; Animals; Asthma; Cell Separation; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Genetic Therapy; Genetic Vectors; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Interleukin-10; Lactococcus lactis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Recombinant Proteins; Th2 Cells

We synthesized a medium consisting of commercial food supplements (food grade medium) that could be used to cultivate Lactobacillus crispatus KT-11 (KT-11), and investigated the antiallergic effects and acute toxicity of KT-11 cultured in this medium. We found that the growth of KT-11 in the food grade medium was comparable to that in DeMan-Rogosa-Sharpe (MRS) medium. Sneezing event was reduced in ovalbumin (OVA)-sensitized BALB/c mice given a diet supplemented with KT-11 grown in the food grade medium (FG-KT-11 group) when compared to mice given a diet supplemented with KT-11 grown in MRS medium (MRS-KT-11 group). The number of CD80(+)CD11b(+) Peyer's patch cells was significantly lower in the FG-KT-11 group than in the MRS-KT-11 group, while IL-12(+)CD11b(+) Peyer's patch cells were higher in the FG-KT-11 group. Only minimal acute toxicity was observed in ICR mice given 1000 or 2000 mg of FG-KT-11/kg body weight. These results suggest that FG-KT-11 represents a safe antiallergic food material.

Topics: Animals; Anti-Allergic Agents; B7-1 Antigen; CD11b Antigen; Cell Count; Culture Media; Dietary Supplements; Female; Hypersensitivity; Interleukin-12; Lactobacillus; Male; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Organ Size; Ovalbumin; Peyer's Patches

The 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a widespread, man-made, persistent organic pollutant with high immunotoxic potentials. It suppresses cell-mediated and humoral immune responses through mechanisms dependent on aryl-hydrocarbon receptor expression and immunosuppressive activity of the cells. Most sensitive to TCDD are organisms during fetal and infant life, mostly due to the developmental stage of many biological systems of the host, including immune system. Recent data show that T regulatory cells that have the potential to suppress immune reactions and which develop after TCDD exposure are also responsible for protection from allergy development. Our goal was to investigate if perinatal exposure to TCDD can affect allergic sensitisation and if T reg cells participate in this phenomenon.. Mice, Balb/c, were perinatally exposed to TCDD or to the carrier. Six weeks old control or exposed mice were sensitised with ovalbumin. Spleen cells of the animals were used to assess the content of T reg cells by means of flow cytometry. Levels of cytokines were assessed by ELISA technique in supernatants of the cells stimulated with anti-CD3 antibody. As a measure of sensitisation, total IgE and anti-OVA IgE were measured in serum of mice by ELISA method. To assess the function of T reg cells isolated from OVA-sensitised control or TCDD exposed animals we performed transfer studies.. Here we show that perinatal exposure to TCDD decreases allergic sensitisation and that this process is related to inhibition of IL-4 synthesis rather than suppression mediated by T regulatory cells.. We hypothesise that dioxin exposure can be an important environmental modulator of immunological responses that participate in allergic reactions.

Topics: Animals; Cytokines; Environmental Pollutants; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Polychlorinated Dibenzodioxins; Pregnancy; Prenatal Exposure Delayed Effects; Spleen; T-Lymphocytes, Regulatory

Type I hypersensitivity is characterized by the overreaction of the immune system against otherwise innocuous substances. It manifests as allergic rhinitis, allergic conjunctivitis, allergic asthma or atopic dermatitis if mast cells are activated in the respective organs. In case of systemic mast cell activation, life-threatening anaphylaxis may occur. Currently, type I hypersensitivities are treated either with glucocorticoids, anti-histamines, or mast cell stabilizers. Although these drugs exert a strong anti-allergic effect, their long-term use may be problematic due to their side-effects.. In the course of a routine in vitro screening process, we identified beta-escin as a potentially anti-allergic compound. Here we tested beta-escin in two mouse models to confirm this anti-allergic effect in vivo. In a model of the early phase of allergic reactions, the murine passive cutaneous anaphylaxis model, beta-escin inhibited the effects of mast cell activation and degranulation in the skin and dose-dependently prevented the extravasation of fluids into the tissue. Beta-escin also significantly inhibited the late response after antigen challenge in a lung allergy model with ovalbumin-sensitized mice. Allergic airway inflammation was suppressed, which was exemplified by the reduction of leucocytes, eosinophils, IL-5 and IL-13 in the bronchoalveolar lavage fluid. Histopathological examinations further confirmed the reduced inflammation of the lung tissue. In both models, the inhibitory effect of beta-escin was comparable to the benchmark dexamethasone.. We demonstrated in two independent murine models of type I hypersensitivity that beta-escin has potent anti-allergic properties. These results and the excellent safety profile of beta-escin suggest a therapeutic potential of this compound for a novel treatment of allergic diseases.

Topics: Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Movement; Cytokines; Dose-Response Relationship, Drug; Eosinophils; Escin; Hypersensitivity; Lung; Mice; Models, Animal; Ovalbumin; Passive Cutaneous Anaphylaxis; Pneumonia; Time Factors; Treatment Outcome

The mice in which the intestinal microflora disruption resulted from antibiotic therapy are challenged by atomization with ovalbumin (OVA) to investigate the relation of allergic airway response and intestinal microflora disruption.. One hundred and twelve female BALB/c mice were divided at random into 6 groups. They were microbiota disruption I group, control I group, microbiota disruption II group, microbiota disruption and challenge group, challenge group and control II group. Cecal contents were collected for quantitative analysis of the intestinal microflora in mice in the former two groups and in mice in the latter four groups on day 6 and day 14, respectively. On day 14, the bronchoalveolar lavage fluid (BALF) was collected for cells counting. OVA-specific IgE in BALF and sera was detected by ELISA. Parts of lungs were collected for histopathology and detection of Th1 and Th2 cell levels by flow of cytometry.. The mice which were given antibiotics suffered from intestinal microbiota disruption. In microbiota disruption and challenge group, eosinophil and lymphocyte infiltration was significant and mucus secretion was increased in lung. The number of total cells, eosinophils, lymphocytes, neutrophils and OVA-specific IgE level were increased in BALF in microbiota disruption and challenge group. Th2 cell levels were increased and Th1 cell levels were not significantly different in microbiota disruption and challenge group compared with those in the control II group.. The allergic (Th2) immune response can be induced by atomization with ovalbumin in the mice in which the intestinal microflora disruption is resulted from antibiotic therapy. The result suggests that the intestinal microflora disruption is a risk factor for allergy and asthma.

Topics: Animals; Anti-Bacterial Agents; Asthma; Bacteria; Disease Models, Animal; Female; Humans; Hypersensitivity; Intestines; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation

Ovalbumin (OVA) is widely used in allergy research. OVA peptide 323-339 has been reported to be responsible for 25-35% of isolated BALB/c mouse T-cell response to intact OVA. An investigation of whether OVA and OVA 323-339 molecules can induce equivalent in vivo and in vitro immune responses was conducted. Eight-week-old BALB/c mice were randomly divided into three groups: OVA, OVA 323-339 and saline. On days 0, 7, 14, mice were intraperitoneally injected with 25 microg OVA or OVA 323-339 absorbed on 300 microg Alum, or saline; on days 21-23, all groups were challenged intranasally with either 20 microl of 1% OVA, 1% OVA 323-339 or saline. On day 28, after killing, splenocytes were isolated and cultured under the stimulus of each allergen or medium. Evaluated by hematoxylin/eosin and major basic protein immunohistochemical stainings, OVA and OVA 323-339 induced similar lung inflammation. Interestingly, significant serum total IgE and OVA-specific IgE were observed in OVA mice when compared to saline control. OVA 323-339 mice showed higher serum OVA-specific IgE, OVA 323-339-specific IgE, IL-4 and lower IFN-gamma similar to OVA mice. The proliferative response to OVA was found in cultured splenocytes of both OVA and OVA 323-339 mice, while the similar proliferative response to OVA 323-339 was only observed in the splenocytes of OVA 323-339-sensitized and challenged mice. Although OVA 323-339 induced a Th2-like response in the mouse model as did OVA, OVA 323-339 has clearly limited immunogenic potency to activate OVA-sensitized and challenged mice splenocytes, unlike OVA.

Topics: Allergens; Animals; Antibody Specificity; Cells, Cultured; Disease Models, Animal; Hypersensitivity; Immunization; Immunodominant Epitopes; Immunoglobulin E; Interferon-gamma; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Peptide Fragments; Pneumonia; Spleen; Th2 Cells

Recent studies highlight the role of Treg in preventing unnecessary responses to allergens and maintaining functional immune tolerance in the lung. We investigated the role of Treg during the sensitization phase in a murine model of experimental allergic airway inflammation by selectively depleting the Treg population in vivo. DEpletion of REGulatory T cells (DEREG) mice were depleted of Treg by diphtheria toxin injection. Allergic airway inflammation was induced using OVA as a model allergen. Pathology was assessed by scoring for differential cellular infiltration in bronchoalveolar lavage, IgE and IgG1 levels in serum, cytokine secretion analysis of lymphocytes from lung draining lymph nodes and lung histology. Use of DEREG mice allowed us for the first time to track and specifically deplete both CD25(+) and CD25(-) Foxp3(+) Treg, and to analyze their significance in limiting pathology in allergic airway inflammation. We observed that depletion of Treg during the priming phase of an active immune response led to a dramatic exacerbation of allergic airway inflammation in mice, suggesting an essential role played by Treg in regulating immune responses against allergens as early as the sensitization phase via maintenance of functional tolerance.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Disease Models, Animal; Forkhead Transcription Factors; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Inflammation; Lung; Lymphocyte Depletion; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory

We have previously demonstrated that intranasal administration of ambient ultrafine particles (UFP) acts as an adjuvant for primary allergic sensitization to ovalbumin (OVA) in Balb/c mice. It is important to find out whether inhaled UFP exert the same effect on the secondary immune response as a way of explaining asthma flares in already-sensitized individuals due to traffic exposure near a freeway. The objective of this study is to determine whether inhalation exposure to ambient UFP near an urban freeway could enhance the secondary immune response to OVA in already-sensitized mice. Prior OVA-sensitized animals were exposed to concentrated ambient UFP at the time of secondary OVA challenge in our mobile animal laboratory in Los Angeles. OVA-specific antibody production, airway morphometry, allergic airway inflammation, cytokine gene expression, and oxidative stress marker were assessed. As few as five ambient UFP exposures were sufficient to promote the OVA recall immune response, including generating allergic airway inflammation in smaller and more distal airways compared with the adjuvant effect of intranasally instilled UFP on the primary immune response. The secondary immune response was characterized by the T helper 2 and IL-17 cytokine gene expression in the lung. In summary, our results demonstrated that inhalation of prooxidative ambient UFP could effectively boost the secondary immune response to an experimental allergen, indicating that vehicular traffic exposure could exacerbate allergic inflammation in already-sensitized subjects.

Topics: Adjuvants, Immunologic; Administration, Inhalation; Animals; Antibody Formation; Asthma; Cytokines; Female; Gene Expression; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Respiratory System; Respiratory Tract Diseases; Severity of Illness Index; Vehicle Emissions

Asthma is a chronic inflammatory disease of the airways characterized by reversible airway obstruction, airway hyperreactivity, and remodeling of the airways. Chlorogenic acid (CGA), an ester of caffeic acid with quinic acid, is one of the most abundant polyphenol compounds in various agricultural products. CGA shows various biological properties, such as anti-oxidant, anti-viral, anti-carcinogenic and anti-inflammatory activities. We investigated suppressive effects of CGA on ovalbumin (OVA)-induced allergic asthma in mice and underlying mechanisms of them. CGA significantly reduced pulmonary eosinophilia and expression of IL-4, IL-5 and TNF-α in the lung as well as the serum levels of total and OVA-specific IgE, while CGA enhanced those of total and OVA-specific IgG3, of which isotype switching is down-regulated by IL-4. In vitro IgE production from LPS/IL-4-stimulated splenocytes was remarkably reduced by CGA, while that of IgG3 was enhanced. The Cε germ line transcription, which is necessary for IL-4 mediated IgE isotype switching, was reduced by CGA in LPS/IL-4-stimulated splenocytes. IgE isotype switching is mediated via several transduction pathways, activating several molecules including STAT-6, NF-κB, ERK1/2, and JNK. Among the molecules, which were activated by IL-4/LPS, activation of STAT-6 and JNK was inhibited by CGA.

Topics: Animals; Artemisia; Asthma; Chlorogenic Acid; Cytokines; Hypersensitivity; Immunoglobulin E; JNK Mitogen-Activated Protein Kinases; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia; STAT6 Transcription Factor; Th2 Cells

Allergies are highly complex disorders with clinical manifestations ranging from mild oral, gastrointestinal, recurrent wheezing, and cutaneous symptoms to life-threatening systemic conditions. The levels of arachidonic acid, eicosanoids, histamine, organic acids and valine are considered to have a variety of physiological functions in connection with allergies. In this research, we have developed a RP-LC/MS method to separate and quantitate six different potential endogenous biomarkers, including leukotriene B(4) (LTB(4)), prostaglandin D(2) (PGD(2)), arachidonic acid (AA), histamine (HI), lactic acid (LA) and valine (VAL), from serum of rats with ovalbumin (OVA)-induced allergy and normal rats, and the discrepancies between the model group and the control group were compared. The separation was performed on a Prevail C18 column (250 mm x 4.6 mm, 5 microm) with a gradient elution of acetonitrile with 0.1% formic acid (v/v) and 10 mM ammonium formate (adjusted to pH 4.0 with formic acid) at a flow rate of 0.5 mL min(-1) The method was validated and shown to be sensitive, accurate (recovery values 76.16-92.57%) and precise (RSD < 10% for all compounds) with a linear range over several orders of magnitude. The method was successfully applied to rat serum and shown to be indicative of the endogenous levels of biomarkers within the rat body. The analysis of the biomarkers can provide insight into the allergic mechanisms associated with related diseases.

Topics: Animals; Arachidonic Acid; Biomarkers; Chromatography, Liquid; Histamine; Hypersensitivity; Lactic Acid; Leukotriene B4; Mass Spectrometry; Ovalbumin; Prostaglandin D2; Rats; Sensitivity and Specificity; Valine

Asthma remains a major cause of morbidity and hospitalizations in developed nations. Despite the widespread prevalence of this disease, the genetic and environmental factors that mediate development and progression of allergic airways disease remain poorly understood. Pulmonary recruitment of eosinophils is believed to contribute to many cardinal features of allergic airways disease. Therefore, it is paramount to understand host factors that contribute to pulmonary eosinophil recruitment into the lungs. Mindin is a component of pulmonary extracellular matrix, which can regulate inflammatory cell recruitment. We characterized the role of mindin in the severity of allergic airways disease using established murine models. There were no baseline differences in wild-type and mindin-deficient animals in cell counts or airway physiology. Using the OVA murine model of allergic airways disease, we observed that mindin-deficient animals have less-severe allergic airways disease with fewer airspace eosinophils and lower lung-lavage levels of inflammatory Th2 cytokines such as IL-13 and IL-4. Furthermore, mindin-deficient animals have reduced airway hyper-responsiveness after methacholine challenge. To determine the role of mindin in eosinophil trafficking, independent of antigen immunization or T lymphocyte activation, we instilled IL-13 directly into the lungs of mice. In this model, mindin regulates eosinophil recruitment into the airspace. In vitro experiments demonstrate that mindin can enhance eotaxin-mediated eosinophil adhesion and migration, which are dependent on the expression of integrins alphaMbeta2 and alpha4beta1. In conclusion, these data suggest that mindin participates in integrin-dependent trafficking of eosinophils and can contribute to the severity of allergic airways disease.

Topics: Animals; Asthma; Cell Adhesion; Cell Movement; Eosinophils; Extracellular Matrix Proteins; Hypersensitivity; Immunoglobulin E; In Vitro Techniques; Integrin alpha4beta1; Interleukin-13; Interleukin-4; Lung; Macrophage-1 Antigen; Methacholine Chloride; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin

We investigated the effects of oral tolerance (OT) in controlling inflammatory response, hyperresponsiveness and airway remodeling in guinea pigs (GP) with chronic allergic inflammation. Animals received seven inhalations of ovalbumin (1-5mg/mL-OVA group) or normal saline (NS group). OT was induced by offering ad libitum ovalbumin 2% in sterile drinking water starting with the 1st ovalbumin inhalation (OT1 group) or after the 4th (OT2 group). The induction of OT in sensitized animals decreased the elastance of respiratory system (Ers) response after both antigen and methacholine challenges, peribronchial edema formation, eosinophilic airway infiltration, eosinophilopoiesis, and airways collagen and elastic fiber content compared to OVA group (P<0.05). The number of mononuclear cells and resistance of respiratory system (Rrs) responses after antigen and methacholine challenges were decreased only in OT2 group compared to OVA group (P<0.05). Concluding, our results show that inducing OT attenuates airway remodeling as well as eosinophilic inflammation and respiratory system mechanics.

Topics: Administration, Inhalation; Airway Resistance; Analysis of Variance; Animals; Chronic Disease; Disease Models, Animal; Dose-Response Relationship, Immunologic; Elastic Tissue; Eosinophils; Extracellular Matrix; Guinea Pigs; Hypersensitivity; Lung; Ovalbumin; Pneumonia; Respiratory Mechanics; Time Factors

Multidrug resistance-associated protein 1 (MRP1) is a cysteinyl leukotriene (CysLT) export pump expressed on mast cells. CysLTs are crucial mediators in allergic airway disease. However, biological significance of MRP1 in allergic airway inflammation has not yet been elucidated. In this study, we sensitized wild-type control mice (mrp1(+/+)) and MRP1-deficient mice (mrp1(-/-)) to ovalbumin (OVA) and challenged them with OVA by aerosol. Airway inflammation and goblet cell hyperplasia after OVA exposure were reduced in mrp1(-/-) mice compared with mrp1(+/+) mice. Furthermore, CysLT levels in bronchoalveolar lavage fluid (BALF) from OVA-exposed mrp1(-/-) mice were significantly lower than those from OVA-exposed mrp1(+/+) mice. Levels of OVA-specific IgE, IL-4, and IL-13 in BALF were also decreased in OVA-exposed mrp1(-/-) mice. IgE-mediated release of CysLTs from murine bone marrow-derived mast cells was markedly impaired by MRP1 deficiency. Our results indicate that MRP1 plays an important role in the development of allergic airway inflammation through regulation of IgE-mediated CysLT export from mast cells.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cysteine; Hypersensitivity; Immunoglobulin E; Leukotrienes; Lung; Mast Cells; Mice; Mice, Inbred Strains; Mice, Mutant Strains; Multidrug Resistance-Associated Proteins; Ovalbumin; Pneumonia

Asthma is a chronic inflammatory disease accompanied by airway obstruction and hyper-responsiveness. Asthmatic inflammation is characterized by the expression of multiple genes for inflammatory mediators. Glycodelin is a glycoprotein with several functions in cell recognition and differentiation. There is substantial evidence that glycodelin may be a mediator for immunomodulatory and immunosuppressive effects on several human tissues. To determine the potential role of glycodelin in the pulmonary immune response, we examined the distribution of the glycodelin mRNA and protein in an experimental rat model of allergen-induced airway inflammation. The experimental model developed an airway response to inhaled nebulized ovalbumin in adult rats. Two groups of rats (ovalbumin and saline) were challenged for 3 weeks, lungs were fixed and embedded, and sections were studied for expression of glycodelin mRNA by in situ hybridization and protein by immunohistochemistry. Glycodelin is expressed in Clara cells of bronchial epithelium, type II pneumocytes and alveolar macrophages. Densitometric analyses show a significant increase of the glycodelin mRNA and protein expression in rat lungs after ovalbumin challenge. Induced glycodelin amounts in tissue, particularly in Clara cells and alveolar macrophages were found. The altered expression pattern of glycodelin may contribute to the pulmonary immune response in asthmatic inflammation.

Topics: Animals; Asthma; Glycoproteins; Hypersensitivity; Inflammation; Lung; Macrophages, Alveolar; Ovalbumin; Pregnancy Proteins; Rats; Respiratory System; RNA, Messenger

Interleukin-13 (IL-13) sequentially binds to IL-13Ralpha1 and IL-4Ralpha forming a high affinity signalling complex. This receptor complex is expressed on multiple cell types in the airway and signals through signal transducer and activator of transcription factor-6 (STAT-6) to stimulate the production of chemokines, cytokines and mucus. Antibodies have been generated, using the UCB Selected Lymphocyte Antibody Method (UCB SLAM), that block either binding of murine IL-13 (mIL-13) to mIL-13Ralpha1 and mIL-13Ralpha2, or block recruitment of mIL-4Ralpha to the mIL-13/mIL-13Ralpha1 complex. Monoclonal antibody (mAb) A was shown to bind to mIL-13 with high affinity (K(D) 11 pM) and prevent binding of mIL-13 to mIL-13Ralpha1. MAb B, that also bound mIL-13 with high affinity (K(D) 8 pM), was shown to prevent recruitment of mIL-4Ralpha to the mIL-13/mIL-13Ralpha1 complex. In vitro, mAbs A and B similarly neutralised mIL-13-stimulated STAT-6 activation and TF-1 cell proliferation. In vivo, mAbs A and B demonstrated equipotent, dose-dependent inhibition of eotaxin generation in mice stimulated by intraperitoneal administration of recombinant mIL-13. In an allergic lung inflammation model in mice, mAbs A and B equipotently inhibited muc5ac mucin mRNA upregulation in lung tissue measured two days after intranasal allergen challenge. These data support the design of therapeutics for the treatment of allergic airway disease that inhibits assembly of the high affinity IL-13 receptor signalling complex, by blocking the binding of IL-13 to IL-13Ralpha1 and IL-13Ralpha2, or the subsequent recruitment of IL-4Ralpha.

Topics: Animals; Antibodies, Blocking; Antibodies, Monoclonal; Cell Line; Cell Line, Tumor; Chemokine CCL11; Disease Models, Animal; Epitopes; Humans; Hypersensitivity; Interleukin-13; Interleukin-13 Receptor alpha1 Subunit; Interleukin-13 Receptor alpha2 Subunit; Male; Mice; Mice, Inbred BALB C; Mucin 5AC; Ovalbumin; Pneumonia; Rabbits; Receptors, Cell Surface; Recombinant Proteins; STAT6 Transcription Factor

Skeletal muscles play key roles in the development of various pathologies, including bronchial asthma and several types of auto-immune disorders, e.g. polymyositis. Since most of these maladies have an immunological/allergic element, this paper is devoted to assessing the impact of immunobiological reorganization on the functional properties of isolated skeletal muscles in mice. A combination of two methods (myography and electrophysiology) was used to evaluate extensor digitorum longus (EDL) and diaphragmatic muscle (DM) in this regard. Conventional myographic technique showed that ovalbumin-induced sensitization (OS) produced different changes in the contractile properties of EDL and DM. The amplitudes of carbachol (CCh)-induced contractions increased in DM but decreased in EDL. Those changes were inversely related to OS-mediated changes of non-quantal acetylcholine (ACh) release intensity within the muscle endplate, as shown by the electrophysiologically measured H-effect. These results clearly show that OS-mediated changes of non-quantal ACh release alter the functional properties of postjunctional ACh receptors and therefore contribute to the disturbance of CCh-induced contractility of skeletal muscles. Other mechanisms of OS-mediated changes of skeletal muscle contractility are also proposed and discussed.

Topics: Acetylcholine; Animals; Carbachol; Cholinergic Agonists; Disease Models, Animal; Electrophysiology; Female; Hypersensitivity; Immunization; Male; Mice; Motor Neurons; Muscle Contraction; Muscle, Skeletal; Myography; Neuromuscular Junction; Ovalbumin; Presynaptic Terminals

Andrographolide - the major active principle isolated from the plant Andrographis paniculata, has been shown to possess a strong anti-inflammatory activity. The possibility that the drug may affect asthmatic inflammation, through inhibition of the relevant inflammatory cytokines, has not been explored. The purpose of this study was, firstly, to investigate the ability of andrographolide to inhibit the release of inflammatory cytokines in vitro in a model of non-specific inflammation and subsequently to determine whether such effect can also be exerted in vivo in allergic lung inflammation. LPS-induced TNF-alpha and GM-CSF release from mouse peritoneal macrophages was inhibited by andrographolide in a concentration-dependent manner. The concentration of the drug producing 50% inhibition was 0.6 microM for TNF-alpha and 3.3 microM for GM-CSF. The maximal inhibition achieved (at 50 microM) was 77% and 94%, respectively, for the two cytokines. The drug was as efficacious as dexamethasone, but about 8-12 times less potent. The drug also suppressed LPS-induced expression of mRNA for the two cytokines, suggesting that this effect may contribute to the mechanism underlying its anti-inflammatory effects. In the in vivo study, intra-peritoneal treatment of ovalbumin-immunized and nasally-challenged mice with andrographolide significantly inhibited the elevation of bronchoalveolar fluid (BAF) levels of TNF-alpha and GM-CSF in a dose-dependent manner, with 30 mg/kg producing an inhibition of 92% and 65% of the cytokines, respectively) and almost completely abolishing the accumulation of lymphocytes and eosinophils. These results provide evidence that andrographolide is an effective anti-inflammatory drug that is active in vitro and in vivo, and affects both non-specific as well as antigen/antibody-dependent lung inflammation. Thus, andrographolide has the potential to be used in a variety of inflammatory conditions, including allergic lung inflammation.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Diterpenes; Granulocyte-Macrophage Colony-Stimulating Factor; Hypersensitivity; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Tumor Necrosis Factor-alpha

Although chronic intestinal helminth infections may suppress allergen-induced airway pathology by inducing a combination of modified T-helper (Th) 2 and immunosuppressive cytokines, a similar capacity of natural acute intestinal infections has remained untested, despite their global prevalence. Here, we show that allergic airway phenotypes including eosinophilia, eotaxin mRNA, and Th2 cytokines are significantly suppressed in animals that were infected by and that have cleared the intestinal parasite Eimeria vermiformis. Unlike in helminth-infected animals, regulation requires temporal coincidence of infection with sensitization; depends on interferon-gamma; and is not associated with an enhanced antigen-specific immunoglobulin G1 response. Moreover, regulation was effective following allergen sensitization in different anatomical sites, and in young and adult mice. These data highlight a transient anatomical dissemination of "functional immunologic dominance" following infection of the gut mucosa. They strongly support the hypothesis that airway allergies are naturally suppressed by both acute and chronic mucosal pathogens, but by different mechanisms.

Topics: Aging; Animals; Chemokine CCL11; Chronic Disease; Coccidiosis; Cytokines; Eimeria; Eosinophilia; Hypersensitivity; Immunization; Immunoglobulin G; Interferon-gamma; Intestinal Mucosa; Mice; Mice, Inbred C57BL; Oocysts; Ovalbumin; Respiratory System; Respiratory Tract Diseases; Th2 Cells

Intra-thoracic antigenic challenge (ovalbumin, 12.5 microg/cavity) led to increased numbers of gammadelta T lymphocytes in pleural cavities, blood and thoracic lymph nodes in sensitized mice within 48 h. Part of these cells expressed CD62L, which increased on gammadelta T cell surfaces obtained from lymph nodes after ovalbumin (OVA) challenge. Selectin blockade by fucoidan pre-treatment (10 mg/kg, i.v.) impaired in vivo increase in CD25(+) and c-fos(+) gammadelta T cell numbers in lymph nodes, indicating a role for selectins on gammadelta T lymphocyte activation and proliferation. In vivo selectin blockade by fucoidan or alpha-CD62L mAb (200 microg/mice, i.p.) also inhibited OVA-induced gammadelta T cell accumulation in pleural cavities. Confirming the direct effect of CD62L on gammadelta T cell transmigration, the migration of i.v. adoptively-transferred CFSE-labeled gammadelta T lymphocytes into pleural cavities of challenged recipient mice was impaired by fucoidan ex vivo treatment. It is noteworthy that eosinophil influx was also impaired in those mice, indicating that reduced eosinophil migration by CD62L in vivo blockade depended on gammadelta T cell migration via CD62L molecules. Accordingly, pleural gammadelta T lymphocytes from fucoidan-treated mice presented reduced OVA-induced IL-5 and CCL11 production. Supporting these data, the depletion of Vgamma4 T lymphocytes, which are pulmonary gammadelta T cells, decreased OVA-induced eosinophil influx into allergic site. Such results demonstrate that CD62L is crucial for the activation of gammadelta T cells in lymph nodes, for their migration into inflamed tissue and for the modulation of eosinophil influx during allergic response.

Topics: Animals; Cell Movement; Cytokines; Eosinophils; Hypersensitivity; L-Selectin; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Pleura; Pleurisy; Polysaccharides; Receptors, Antigen, T-Cell, gamma-delta

The impact of genetic factors on asthma is well recognized but poorly understood. We tested the hypothesis that different mouse strains present different lung tissue strip mechanics in a model of chronic allergic asthma and that these mechanical differences may be potentially related to changes of extracellular matrix composition and/or contractile elements in lung parenchyma. Oscillatory mechanics were analysed before and after acetylcholine (ACh) in C57BL/10, BALB/c, and A/J mice, subjected or not to ovalbumin sensitization and challenge. In controls, tissue elastance (E) and resistance (R), collagen and elastic fibres' content, and alpha-actin were higher in A/J compared to BALB/c mice, which, in turn, were more elevated than in C57BL/10. A similar response pattern was observed in ovalbumin-challenged animals irrespective of mouse strain. E and R augmented more in ovalbumin-challenged A/J [E: 22%, R: 18%] than C57BL/10 mice [E: 9.4%, R: 11%] after ACh In conclusion, lung parenchyma remodelled differently yielding distinct in vitro mechanics according to mouse strain.

Topics: Animals; Asthma; Chronic Disease; Disease Models, Animal; Extracellular Matrix; Hypersensitivity; In Vitro Techniques; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Respiratory Mechanics; Species Specificity

The differentiation of T cells along different lineages is central to the control of immunity. Here we have used a conditional gene knockout system to delete PKC lambda/iota selectively in activated T cells. With this system we have demonstrated that PKC lambda/iota is necessary for T-helper cell (Th2) cytokine production and optimal T-cell proliferation and allergic airway inflammation in vivo. Our data demonstrate that the activation of the transcription factors nuclear factor of activated T cells and NF-kappaB is impaired in PKC lambda/iota-deficient activated T cells. In addition, we present genetic knockout evidence in ex vivo experiments with primary T cells that PKC lambda/iota is critical for the control of cell polarity during T-cell activation. Therefore PKC lambda/iota emerges as a critical regulator of Th 2 activation.

Topics: Animals; Cell Differentiation; Cell Polarity; Cell Proliferation; Cytokines; Hypersensitivity; Immunoglobulin E; Inflammation; Isoenzymes; Lymphocyte Activation; Mice; Mice, Knockout; Ovalbumin; Protein Kinase C; Respiratory System; Th2 Cells; Transcription Factors; Up-Regulation

Endothelial lipase (EL) is a novel phospholipase that determines plasma high-density lipoprotein cholesterol (HDL-C) levels. We have investigated the role of HDL-C in lung allergic inflammation by using EL knockout (EL-KO) mice that are high in HDL-C. EL-KO and wild-type control mice were sensitized and challenged with ovalbumin to evoke eosinophilic inflammation in the lung. EL was expressed in epithelial cells, alveolar type II cells, and endothelial cells in the lung, and its expression was upregulated during inflammation. Concomitant with attenuated hyperresponsiveness of the airway smooth muscles, the number of eosinophils in bronchoalveolar lavage and the expression of VCAM-1 were lower in EL-KO mice than in control mice. HDL reduced cytokine-induced VCAM-1 expression in cultured endothelial cells. When plasma HDL levels were decreased to similar levels in both mouse groups by adenovirus-mediated overexpression of EL, however, eosinophil infiltration was still lower in EL-KO mice. In vitro adhesion assays revealed that EL expression on the cell surface promoted the interaction of eosinophils through the ligand-binding function of EL. In summary, targeted inactivation of EL attenuated allergic inflammation in the lung, and the protective effects in EL-KO mice were associated with high plasma HDL levels, downregulation of VCAM-1, and loss of the direct ligand-binding function of EL. Thus EL is a novel modulator of the progression of allergic asthma.

Topics: Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Adhesion; Cell Movement; Chlorocebus aethiops; Cholesterol, HDL; COS Cells; Endothelium; Eosinophils; Gene Targeting; Humans; Hypersensitivity; Lipase; Lung; Mice; Mice, Knockout; Ovalbumin; Pneumonia; Vascular Cell Adhesion Molecule-1

Gene silencing using small interfering RNA (siRNA) is a potent method of specifically knocking down molecular targets. Small interfering RNA is therapeutically promising, however, treatment of allergic diseases with siRNA has not been explored in vivo. The aim of this study was to evaluate therapeutic effects of CD40 siRNA on inhibition of allergic responses.. Mice sensitized with ovalbumin (OVA) and alum were treated with CD40 siRNA, scrambled siRNA, or phosphate buffer saline (PBS) alone, and then challenged intranasally with OVA.. A significant reduction in nasal allergic symptoms was observed in the CD40 siRNA treated OVA-allergic mice compared to the controls of scrambled siRNA and PBS alone, which is correlated with the decrease of local eosinophil accumulation. CD40 siRNA treatment knocked down CD40 expression on dendritic cells (DCs) in vivo and impaired their antigen presenting function. Treatment with CD40 siRNA resulted in inhibition of OVA-specific T cell response and decrease of interleukin-4 (IL-4), IL-5, and interferon-gamma production from T cells stimulated with OVA. Administration of CD40 siRNA also suppressed CD40 expression on B cells, resulting in down-regulation of OVA-specific immunoglobulin E (IgE), IgG1, and IgG2a levels. Additionally, increased regulatory T cells were observed in the CD40 siRNA treated mice.. The present study demonstrates a novel therapeutic use for siRNA in allergy. CD40 siRNA attenuated allergy through inhibition of DC and B cell functions and generation of regulatory T (Treg) cells.

Topics: Allergens; Animals; B-Lymphocytes; CD40 Antigens; Dendritic Cells; Gene Silencing; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; T-Lymphocytes, Regulatory

Interactions between T cells and dendritic cells in the airway mucosa precede secondary immune responses to inhaled antigen. The purpose of this study was to identify the anatomical locations where dendritic cell-T cell interactions occur, resulting in T cells activation by dendritic cells. In a mouse model of allergic airway inflammation, we applied whole-mount immunohistology and confocal microscopy to visualize dendritic cells and T cells together with nerves, epithelium, and smooth muscle in three dimensions. Proliferating T cells were identified by the detection of the incorporation of the nucleotide analogue 5-ethynyl-2'-deoxyuridine into the DNA. We developed a novel quantification method that enabled the accurate determination of cell-cell contacts in a semi-automated fashion. Dendritic cell-T cell interactions occurred beneath the smooth muscle layer, but not in the epithelium. Approximately 10% of the dendritic cells were contacted by nerves, and up to 4% of T cells formed clusters with these dendritic cells. T cells that were clustered with nerve-contacting dendritic cells proliferated only in the airways of mice with allergic inflammation but not in the airways of negative controls. Taken together, these results suggest that during the secondary immune response, sensory nerves influence dendritic cell-driven T cell activation in the airway mucosa.

Topics: Animals; CD11c Antigen; Cell Division; Dendritic Cells; Disease Models, Animal; Hypersensitivity; Inflammation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Confocal; Nerve Fibers; Neurons; Ovalbumin; Respiratory System; T-Lymphocytes

Recent clinical trials, epidemiological studies and animal experiments have suggested that probiotics may help suppress the development of allergic responses.. To investigate whether the application of the probiotic Escherichia coli strain Nissle 1917 (EcN) protects mice from developing ovalbumin (OVA)-specific T helper-2 responses in the airways.. OVA-specific Th2 responses were induced by 2 intraperitoneal (i.p.) injections with OVA/alum followed by 1 intranasal (i.n.) challenge with OVA. EcN was given orally during the entire sensitization and challenge period, together with OVA/alum during the i.p. sensitizations, or i.n. before or during the airway challenge with OVA.. We found that when the bacteria were given together with OVA/alum airway eosinophilia, airway hyper-reactivity, goblet cell metaplasia and IL-5 levels in the bronchoalveolar lavage and mediastinal lymph node cell cultures were reduced. This effect was associated with increased numbers of IFN-gamma producing T helper-1 cells and IFN-gamma levels in the airways and strongly increased OVA-specific IgG(2a) titers in the serum. The suppressive effect on airway eosinophilia was dependent on IFN-gamma but not TLR-4. Applying EcN i.n. or orally did not reduce the development of allergen-specific Th2 responses.. Our results suggest that EcN can inhibit the development of allergic responses when the bacteria are present at the site of Th2 cell priming and that this immunomodulatory effect is due to a shift from Th2 to Th1 response. The data support the hypothesis that probiotics may help reduce allergic responses and that EcN may also be used as adjuvant therapy to induce allergen-specific Th1 responses.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Administration, Oral; Allergens; Alum Compounds; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Dendritic Cells; Eosinophilia; Escherichia coli; Female; Goblet Cells; Hypersensitivity; Interferon-gamma; Interleukin-5; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Ovalbumin; Probiotics; Th1 Cells; Th2 Cells

Sublingual immunotherapy (SLIT) efficacy could be improved by formulations facilitating allergen contact with the oral mucosa and uptake by antigen-presenting cells (APCs).. Two types of chitosan microparticles, differing in size and surface charge, were tested in vitro for their capacity to improve antigen uptake and presentation by murine bone marrow-derived dendritic cells (BMDCs) or purified oral APCs. T-cell priming in cervical lymph nodes (LNs) was assessed by intravenous transfer of carboxyfluorescein diacetate succinimidyl ester-labelled ovalbumin (OVA)-specific CD4+ T cells and flow cytometry analysis. Ovalbumin-sensitized BALB/c mice were treated sublingually with soluble or chitosan-formulated OVA twice a week for 2 months. Airway hyperresponsiveness (AHR), lung inflammation and T-cell responses in cervical and mediastinal LNs were assessed by whole-body plethysmography, lung histology and Cytometric Bead Array technology, respectively.. Only a mucoadhesive (i.e. highly positively charged) and microparticulate form of chitosan enhances OVA uptake, processing and presentation by murine BMDCs and oral APCs. Targeting OVA to dendritic cells with this formulation increases specific T-cell proliferation and IFN-gamma/IL-10 secretion in vitro, as well as T-cell priming in cervical LNs in vivo. Sublingual administration of such chitosan-formulated OVA particles enhances tolerance induction in mice with established asthma, with a dramatic reduction of both AHR, lung inflammation, eosinophil numbers in bronchoalveolar lavages, as well as antigen-specific Th2 responses in mediastinal LNs.. Mucoadhesive chitosan microparticles represent a valid formulation for sublingual allergy vaccines.

Topics: Administration, Sublingual; Adoptive Transfer; Animals; Antigen Presentation; CD4-Positive T-Lymphocytes; Chelating Agents; Chitosan; Dendritic Cells; Desensitization, Immunologic; Female; Hypersensitivity; Immune Tolerance; Interferon-gamma; Interleukin-10; Lung; Mice; Mice, Inbred BALB C; Mouth Mucosa; Ovalbumin; Th2 Cells

We recently demonstrated that the T-helper type 1 (Th1) immune response plays an important role in the development of non-eosinophilic inflammation induced by airway exposure of an allergen plus double-stranded RNA (dsRNA). However, the role of lipoxygenase (LO) metabolites in the development of Th1 inflammation is poorly understood.. To evaluate the role of LO metabolites in the development of Th1 inflammation induced by sensitization with an allergen plus dsRNA.. A Th2-allergic inflammation mouse model was created by an intraperitoneal injection of lipopolysaccharide-depleted ovalbumin (OVA, 75 microg) and alum (2 mg) twice, and the Th1 model was created by intranasal application of OVA (75 microg) and synthetic dsRNA [10 microg of poly(I : C)] four times, followed by an intranasal challenge with 50 microg of OVA four times. The role of LO metabolites was evaluated using two approaches: a transgenic approach using 5-LO(-/-) and 15-LO(-/-) mice, and a pharmacological approach using inhibitors of cysteinyl leucotriene receptor-1 (cysLTR1), LTB4 receptor (BLT1), and 15-LO.. We found that the Th1-allergic inflammation induced by OVA+dsRNA sensitization was similar between 5-LO(-/-) and wild-type (WT) control mice, although Th2 inflammation induced by sensitization with OVA+alum was reduced in the former group. In addition, dsRNA-induced Th1 allergic inflammation, which is associated with down-regulation of 15-hydroxyeicosateraenoic acids production, was not affected by treatment with cysLTR1 or BLT1 inhibitors, whereas it was significantly lower in 12/15-LO(-/-) mice compared with WT control mice. Moreover, dsRNA-induced allergic inflammation and the recruitment of T cells following an allergen challenge were significantly inhibited by treatment with a specific 15-LO inhibitor (PD146176).. 15-LO metabolites appear to be important mediators in the development of Th1-allergic inflammation induced by sensitization with an allergen plus dsRNA. Our findings suggest that the 15-LO pathway is a novel therapeutic target for the treatment of virus-associated asthma characterized by Th1 inflammation.

Topics: Acetates; Allergens; Alum Compounds; Animals; Arachidonate 15-Lipoxygenase; Arachidonate 5-Lipoxygenase; Cyclopropanes; Disease Models, Animal; Fatty Alcohols; Fluorenes; Glycols; Hypersensitivity; Inflammation; Leukotriene Antagonists; Lipoxygenase Inhibitors; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Poly I-C; Quinolines; Receptors, Leukotriene; Receptors, Leukotriene B4; RNA, Double-Stranded; Sulfides; Th1 Cells; Th2 Cells

The adjuvant effect of particles on allergic immune responses has been shown to increase with decreasing particle size and increasing particle surface area. Like ultrafine particles, carbon nanotubes (CNTs) have nano-sized dimensions and a large relative surface area and might thus increase allergic responses. Therefore, we examined whether single-walled (sw) and multi-walled (mw) CNTs have the capacity to promote allergic responses in mice, first in an sc injection model and thereafter in an intranasal model. Balb/cA mice were exposed to three doses of swCNT, mwCNT, as well as ultrafine carbon black particles (ufCBPs, Printex90) during sensitization with the allergen ovalbumin (OVA). Five days after an OVA booster, OVA-specific IgE, IgG1, and IgG2a antibodies in serum and the numbers of inflammatory cells and cytokine levels in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, ex vivo OVA-induced cytokine release from mediastinal lymph node (MLN) cells was measured. In separate experiments, differential cell counts were determined in BALF 24 h after a single intranasal exposure to the particles in the absence of allergen. We demonstrate that both swCNT and mwCNT together with OVA strongly increased serum levels of OVA-specific IgE, the number of eosinophils in BALF, and the secretion of Th2-associated cytokines in the MLN. On the other hand, only mwCNT and ufCBP with OVA increased IgG2a levels, neutrophil cell numbers, and tumor necrosis factor-alpha and monocyte chemoattractant protein-1 levels in BALF, as well as the acute influx of neutrophils after exposure to the particles alone. This study demonstrates that CNTs promote allergic responses in mice.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Analysis of Variance; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Injections, Subcutaneous; Lymph Nodes; Mice; Mice, Inbred BALB C; Microscopy, Electron; Nanotubes, Carbon; Ovalbumin; Soot

In healthy individuals, humoral immune responses to allergens consist of serum IgA and IgG4, whereas cellular immune responses are controlled by regulatory T (Treg) cells. In search of new compounds that might prevent the onset of allergies by stimulating this type of immune response, we have focused on the mucosal adjuvant, cholera toxin B (CTB), as it induces the formation of Treg cells and production of IgA. Here, we have found that CTB suppresses the potential of dendritic cells to prime for Th2 responses to inhaled allergen. When we administered CTB to the airways of naïve and allergic mice, it strongly suppressed the salient features of asthma, such as airway eosinophilia, Th2 cytokine synthesis, and bronchial hyperreactivity. This beneficial effect was only transferable to other mice by transfer of B but not of T lymphocytes. CTB caused a transforming growth factor-beta-dependent rise in antigen-specific IgA in the airway luminal secretions, which was necessary for its preventive and curative effect, as all effects of CTB were abrogated in mice lacking the luminal IgA transporting polymeric Ig receptor. Not only do these findings show a novel therapeutic avenue for allergy, they also help to explain the complex relationship between IgA levels and risk of developing allergy in humans.

Topics: Adjuvants, Immunologic; Adoptive Transfer; Allergens; Animals; B-Lymphocytes; Cholera Toxin; Cytokines; Dendritic Cells; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin A, Secretory; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells; Transforming Growth Factor beta

Sensitization of esophageal sensory afferents by inflammatory mediators plays an important role in esophageal nociception. We have shown esophageal mast cell activation induces long-lasting mechanical hypersensitivity in vagal nodose C-fibers. However, the roles of mast cell mediators and downstream ion channels in this process are unclear. Mast cell tryptase via protease-activated receptor 2 (PAR2)-mediated pathways sensitizes sensory nerves and induces hyperalgesia. Transient receptor potential A1 (TRPA1) plays an important role in mechanosensory transduction and nociception. Here we tested the hypothesis that mast cell activation via a PAR2-dependent mechanism sensitizes TRPA1 to induce mechanical hypersensitivity in esophageal vagal C-fibers. The expression profiles of PAR2 and TRPA1 in vagal nodose ganglia were determined by immunostaining, Western blot, and RT-PCR. Extracellular recordings from esophageal nodose neurons were performed in ex vivo guinea pig esophageal-vagal preparations. Action potentials evoked by esophageal distention and chemical perfusion were compared. Both PAR2 and TRPA1 expressions were identified in vagal nodose neurons by immunostaining, Western blot, and RT-PCR. Ninety-one percent of TRPA1-positive neurons were of small and medium diameters, and 80% coexpressed PAR2. Esophageal mast cell activation significantly enhanced the response of nodose C-fibers to esophageal distension (mechanical hypersensitivity). This was mimicked by PAR2-activating peptide, which sustained for 90 min after wash, but not by PAR2 reverse peptide. TRPA1 inhibitor HC-030031 pretreatment significantly inhibited mechanical hypersensitivity induced by either mast cell activation or PAR2 agonist. Collectively, our data provide new evidence that sensitizing TRPA1 via a PAR2-dependent mechanism plays an important role in mast cell activation-induced mechanical hypersensitivity of vagal nodose C-fibers in guinea pig esophagus.

Topics: Afferent Pathways; Animals; Disease Models, Animal; Esophagus; Evoked Potentials; Guinea Pigs; Hyperalgesia; Hypersensitivity; Male; Mast Cells; Mechanotransduction, Cellular; Nerve Fibers, Unmyelinated; Nodose Ganglion; Ovalbumin; Physical Stimulation; Pressure; Receptor, PAR-2; Stimulation, Chemical; Stress, Mechanical; Transient Receptor Potential Channels; Tryptases; Vagus Nerve

Leuconostoc citreum (L. citreum) HJ-P4 (KACC 91035) is one of the major predominant species in kimchi fermentation in Korea. The purpose of the present study was to test the immunomodulatory capacity of L. citreum to modulate the IgE-mediated allergic response and to examine the involvement of NF-kappaB and MAPK in IL-12 production in macrophages. Balb/c mice were sensitized with OVA/alum and oral administration of L. citreum to the mice began before or after the OVA sensitization. Protein and mRNA expression of Th1 cytokines in splenocytes by L. citreum in vitro was measured. The role of NF-kappaB and MAPK such as p38, ERK1/2 and JNK in L. citreum-induced IL-12 was investigated in peritoneal macrophages and RAW264.7 cell lines. L. citreum inhibited the serum levels of total IgE, IgG1 and IgG2a altogether and increased OVA-specific IFN-gamma production in splenocytes from pre- and post-sensitized animals. However, the downregulation of IL-4 and IL-5 production was observed only in the pre-sensitization group. The ability of L. citreum to stimulate IFN-gamma was dependent on its induction of IL-12. NF-kappaB, p38 and JNK were mainly involved in L. citreum-induced IL-12 production. In conclusion, the current study demonstrated that L. citreum is able to regulate serum IgE generation at the induction and effector phases of allergic response through overall control over antibody production and that its involvement of IL-12 production was mediated through NF-kappaB and p38/JNK. Taken together, the use of L. citreum can be useful in preventing the development and progression of IgE production.

Topics: Animals; Cell Line; Cells, Cultured; Cytokines; Disease Models, Animal; Fermentation; Humans; Hypersensitivity; Immunoglobulin E; Interleukin-12; JNK Mitogen-Activated Protein Kinases; Korea; Leuconostoc; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Signal Transduction; Th1 Cells; Vegetables

Epidemiological and clinical evidence has suggested that increased dietary intake of fish oil containing omega-3 fatty acids including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) may be associated with a reduced risk of asthma. However, interventional studies on these effects have been equivocal and controversial. Free radical oxidation products of lipids and cyclooxygenases-derived prostaglandins are believed to play an important role in asthma, and fish oil supplementation may modulate the levels of these critical lipid mediators. We employed a murine model of allergic inflammation produced by sensitization to ovalbumin (OVA) to study the effects of fish oil supplementation on airway inflammation. Our studies demonstrated that omega-3 fatty acids were dose dependently incorporated into mouse lung tissue after dietary supplementation. We examined the oxidative stress status by measuring the levels of isoprostanes (IsoPs), the gold standard for oxidative stress in vivo. OVA challenge caused significant increase of F(2)-IsoPs in mouse lung, suggesting an elevated level of oxidative stress. Compared to the control group, fish oil supplementation led to a significant reduction of F(2)-IsoP (from arachidonic acid) with a concomitant increase of F(3)-IsoPs (from EPA) and F(4)-IsoPs (from DHA). Surprisingly, however, fish oil supplementation enhanced production of proinflammatory cytokine IL-5 and IL-13. Furthermore, fish oil supplementation suppressed the production of pulmonary protective PGE(2) in the bronchoalveolar lavage (BAL) while the level of urinary metabolites of the PGE(2) was increased. Our data suggest that augmented lung inflammation after fish oil supplementation may be due to the reduction of PGE(2) production in the lung and these dichotomous results bring into question the role of fish oil supplementation in the treatment of asthma.

Topics: Animals; Body Composition; Cytokines; Dietary Supplements; Down-Regulation; F2-Isoprostanes; Fatty Acids, Omega-3; Female; Fish Oils; Hypersensitivity; Inflammation Mediators; Lipid Metabolism; Mice; Mice, Inbred BALB C; Models, Biological; Ovalbumin; Pneumonia

Naturally occurring Foxp3(+)CD4(+)CD25(+) T cells isolated from lungs of naive mice regulate lung allergic airway hyperresponsiveness, inflammation, levels of Th2 cytokines, and mucus production. OVA-specific (alphabetaTCR(+)) CD4(+)CD25(+) T cells suppressed ragweed-induced airway hyperresponsiveness and inflammation as did anti-TCR-treated OVA-specific CD4(+)CD25(+) T cells, suggesting that Ag-specificity was not required for expression of regulatory activities. Suppression was associated with increased levels of IL-10 and TGF-beta; decreased levels of IL-4, IL-5, and IL-13 in bronchoalveolar lavage fluid; and reduced recruitment and activation of CD8(+) T cells in the airways. Following intratracheal administration, OVA-specific CD4(+)CD25(+) T cells were identified in both the airway lumens and lung parenchyma, and in some instances in close proximity to host CD8(+) T cells. These results demonstrate that the regulatory activities of naturally occurring Foxp3(+)CD4(+)CD25(+) T cells on lung allergic responses are Ag-nonspecific and thus, independent of Ag-specific recognition.

Topics: Adoptive Transfer; Ambrosia; Animals; CD8-Positive T-Lymphocytes; Cytokines; Epitopes; Hypersensitivity; Inflammation; Lung Diseases; Mice; Mice, Inbred Strains; Ovalbumin; T-Lymphocytes, Regulatory

The effects of ultrafine and fine particles of zinc oxide (ZnO) on IgE-dependent mast cell activation were investigated. The rat mast cell line RBL2H3 sensitized with monoclonal anti-ovalbumin (OVA) IgE was challenged with OVA in the presence or absence of ZnO particles and zinc sulfate (ZnSO4). Degranulation of RBL2H3 was examined by the release of β-hexosaminidase. To understand the mechanisms responsible for regulating mast cell functions, the effects of ZnO particles on the levels of intracellular Zn2+, Ca2+, phosphorylated-Akt, and global tyrosine phosphorylation were also measured. IgE-induced release of b-hexosaminidase was obviously attenuated by ultrafine ZnO particles and ZnSO4, whereas it was very weakly inhibited by fine ZnO particles. The intracellular Zn2+ concentration was higher in the cells incubated with ultrafine ZnO particles than in those with fine ZnO particles. Consistent with inhibitory effect on release of b-hexosaminidase, ultrafine ZnO particles and ZnSO4, but not fine ZnO particle, strongly attenuated the IgE-mediated increase of phosphorylated-Akt and tyrosine phosphorylations of 100 and 70 kDa proteins in RBL2H3 cells. These findings indicate that ultrafine ZnO particles, with a small diameter and a large total surface area/mass, could release Zn2+ easily and increase intracellular Zn2+ concentration efficiently, thus decreasing FceRI-mediated mast cell degranulation through inhibitions of PI3K and protein tyrosine kinase activation. Exposure to ZnO particles might affect immune responses, especially in allergic diseases.

Topics: Animals; beta-N-Acetylhexosaminidases; Calcium; Cell Degranulation; Cell Line; Hypersensitivity; Immunoglobulin E; Mast Cells; Ovalbumin; Particle Size; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Protein-Tyrosine Kinases; Proto-Oncogene Proteins c-akt; Rats; Receptors, IgE; Signal Transduction; Tyrosine; Zinc; Zinc Oxide

We showed recently that IL-4 causes mitochondrial dysfunction in allergic asthma. IL-4 is also known to induce 12/15-lipoxygenase (12/15-LOX), a potent candidate molecule in asthma. Because vitamin E (Vit-E) reduces IL-4 and inhibits 12/15-LOX in vitro, here we tested the hypothesis that Vit-E may be effective in restoring key mitochondrial dysfunctions, thus alleviating asthma features in an experimental allergic murine model. Ovalbumin (OVA)-sensitized and challenged male BALB/c mice showed the characteristic features of asthma such as airway hyperresponsiveness (AHR), airway inflammation, and airway remodeling. In addition, these mice showed increase in the expression and metabolites of 12/15-LOX, reduction in the activity and expression of the third subunit of mitochondrial cytochrome-c oxidase, and increased cytochrome c in lung cytosol, which indicate that OVA sensitization and challenge causes mitochondrial dysfunction. Vit-E was administered orally to these mice, and 12/15-LOX expression, key mitochondrial functions, ultrastructural changes of mitochondria in bronchial epithelia, and asthmatic parameters were determined. Vit-E treatment reduced AHR, Th2 response including IL-4, IL-5, IL-13, and OVA-specific IgE, eotaxin, transforming growth factor-beta1, airway inflammation, expression and metabolites of 12/15-LOX in lung cytosol, lipid peroxidation, and nitric oxide metabolites in the lung, restored the activity and expression of the third subunit of cytochrome-c oxidase in lung mitochondria and bronchial epithelia, respectively, reduced the appearance of cytochrome c in lung cytosol, and also restored mitochondrial ultrastructural changes of bronchial epithelia. In summary, these findings show that Vit-E reduces key mitochondrial dysfunctions and alleviates asthmatic features.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Airway Remodeling; Animals; Anti-Asthmatic Agents; Arachidonate 12-Lipoxygenase; Arachidonate 15-Lipoxygenase; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Cytochromes c; Disease Models, Animal; Electron Transport Complex IV; Goblet Cells; Hyperplasia; Hypersensitivity; Immunoglobulin E; Interleukin-13; Interleukin-4; Interleukin-5; Linoleic Acids; Lung; Male; Mice; Mice, Inbred BALB C; Mitochondria; Ovalbumin; Oxidative Stress; Pulmonary Fibrosis; Transforming Growth Factor beta1; Vitamin E

Asthma pathogenesis involves gene and environmental interactions. A disintegrin and metalloprotease 33 (ADAM33)/Adam33 is a susceptibility gene for asthma and bronchial hyperresponsiveness in human beings and mice. ADAM33 is almost exclusively expressed in mesenchymal cells, including mesenchymal progenitors in developing lungs.. Because maternal allergy is a risk factor for asthma, we hypothesized that an allergic environment affects ADAM33/Adam33 expression during human and mouse lung development.. Human embryonic/fetal lung (HEL) tissues were collected from first-trimester terminations of pregnancy. These were processed immediately or used for explant culture +/- IL-13. MF1 mice or ovalbumin-sensitized A/J mice (Bronchial hyperresponsivness (Bhr)1/Adam33 locus-positive) were time-mated and challenged with ovalbumin (A/J mice only) during pregnancy. Lungs were harvested at different times during gestation and post partum. ADAM33/Adam33 expression was analyzed by using reverse transcriptase quantitative polymerase chain reaction and Western blotting.. ADAM33 mRNA was detectable in HELs in the pseudoglandular stage of development and showed a significant increase from 7 to 9 weeks postconception. IL-13 significantly suppressed ADAM33 mRNA in HEL explants. In developing murine lungs, Adam33 mRNA and protein expression increased significantly in the early pseudoglandular stage and showed another large increase post partum. In A/J mice, maternal allergy significantly suppressed Adam33 mRNA in lungs of newborn pups, whereas processed Adam33 protein increased and several smaller isoforms were detected.. Adam33/Adam33 shows 2 significant increments in expression during lung morphogenesis, suggesting important developmental regulation. The ability of maternal allergy or exogenous IL-13 to suppress Adam33/ADAM33 mRNA but enhance Adam33 processing suggests a gene-environment interaction that may be relevant for asthma pathogenesis.

Topics: ADAM Proteins; Animals; Gene Expression Regulation, Developmental; Humans; Hypersensitivity; Interleukin-13; Lung; Mice; Organ Culture Techniques; Ovalbumin; Protein Isoforms; RNA, Messenger

Prospective cohort studies suggest that children hospitalized in early life with severe infections are significantly more likely to develop recurrent wheezing and asthma.. Using an inhalational mouse model of allergic airways inflammation, we sought to determine the effect of viral and bacterial-associated molecular patterns on the magnitude of the allergic inflammatory response and whether this effect was age dependent.. BALB/c mice were sensitized by intranasal administration of endotoxin(low) ovalbumin (OVA) in the absence or presence of viral single-stranded (ss)RNA, lipoteichoic acid or flagellin as neonates (within the first 24 h of life) or as weanlings (4 weeks of age). Mice were challenged four times with OVA at 6 weeks of age and end-points (bronchoalveolar lavage cytology, histology, antigen-specific T and B cell responses) determined at 7 weeks of age.. Inhalational sensitization (<24 h or 4 weeks of age) and challenge with OVA induced a mild allergic inflammatory response in the airways as indicated by increased numbers of eosinophils and mucus cells, elevated serum OVA-specific IgG1, and production of T helper 2 (Th2) cytokines. Mice sensitized to endotoxin(low) OVA at birth in the presence of ssRNA or lipoteichoic acid, but not flagellin, showed an increase in the numbers of airway and tissue eosinophils, mucus producing cells and antigen-specific production of IL-13 as compared with mice exposed only to endotoxin(low) OVA. By contrast, all three TLR ligands failed to increase the magnitude of OVA-induced allergic inflammation in mice sensitized as weanlings.. Recognition of distinct microbial-associated patterns in early life may preferentially promote the de novo differentiation of bystander, antigen-specific CD4(+) T cells toward a Th2 phenotype, and promote an asthma-like phenotype upon cognate antigen exposure in later life.

Topics: Adjuvants, Immunologic; Animals; Animals, Newborn; Eosinophilia; Flagellin; Gene Expression; Hyperplasia; Hypersensitivity; Immunoglobulin G; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Lipopolysaccharides; Lung; Lymph Nodes; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mucous Membrane; Ovalbumin; RNA, Viral; Teichoic Acids; Th2 Cells; Toll-Like Receptor 2; Toll-Like Receptor 7; Vaccination

Allergens can be maternally transferred to the fetus or neonate, though it is uncertain how this initial allergen exposure may impact the development of allergy responses. To evaluate the roles of timing and level of maternal allergen exposure in the early life sensitization of progeny, female BALB/c mice were given ovalbumin (OVA) orally during pregnancy, lactation or weekly at each stage to investigate the immunoglobulin E (IgE) antibody production and cellular responsiveness of their offspring. Exposure to OVA during pregnancy was also evaluated in OVA-specific T-cell receptor (TCR) transgenic (DO11.10) mice. The effect of prenatal antigen exposure on offspring sensitization was dependent on antigen intake, with low-dose OVA inducing tolerance followed by neonatal immunization that was sustained even when pups were immunized when 3 weeks old. These offspring received high levels of transforming growth factor-beta via breastfeeding. High-dose exposure during the first week of pregnancy or perinatal period induced transient inhibition of IgE production following neonatal immunization; although for later immunization IgE production was enhanced in these offspring. Postnatal maternal antigen exposure provided OVA transference via breastfeeding, which consequently induced increased offspring susceptibility to IgE antibody production according to week post-birth. The effect of low-dose maternal exposure during pregnancy was further evaluated using OVA transgenic TCR dams as a model. These progeny presented pronounced entry of CD4(+) T cells into the S phase of the cell cycle with a skewed T helper type 2 response early in life, revealing the occurrence of allergen priming in utero. The balance between tolerance and sensitization depended on the amount and timing of maternal allergen intake during pregnancy.

Topics: Administration, Oral; Allergens; Animals; CD4-Positive T-Lymphocytes; Cytokines; Female; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Male; Maternal Exposure; Maternal-Fetal Exchange; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Pregnancy; Receptors, Antigen, T-Cell; Transforming Growth Factor beta

Dendritic cells (DCs) are essential for the initiation and maintenance of T(H)2 responses to inhaled antigen that lead to the establishment of allergic diseases. Two subpopulations of nonplasmacytoid DCs (ie, CD11b(low)CD103+ and CD11b(high)CD103(-)) are found in lung/airway tissues. Yet the identification and migratory properties of the DC subset that contributes to T(H)2-mediated responses remain to be clarified. CD47, a signal regulatory protein (SIRP)-alpha partner, reportedly governed skin DC migration.. We here thought to investigate the role of CD47/SIRP-alpha interactions in airway DC trafficking and the development of allergic airway inflammation.. We characterized the DC influx into lungs and mediastinal lymph nodes in CD47(-/-) and CD47(+/+) BALB/c mice by using experimental models of allergic asthma. Mice were systemically (intraperitoneal ovalbumin/alum) or locally (intratracheal ovalbumin-loaded bone marrow-derived DCs) immunized and challenged by ovalbumin aerosol. We also evaluated the consequences of SIRP-alpha-Fc fusion molecule administration on the induction of airway disease in BALB/c mice.. SIRP-alpha selectively identified the CD11b(high)CD103(-) DC subset that predominantly accumulated in mediastinal lymph nodes during airway inflammation. However, CD103(-)SIRP-alpha+ DC trafficking, T(H)2 responses, and airway disease were impaired in CD47(-/-) mice. Importantly, the adoptive transfer of CD103(-) SIRP-alpha+CD47(+/+) but not CD47(-/-) DCs elicited a strong T(H)2 response in CD47(-/-) mice. Finally, the administration of SIRP-alpha-Fc molecule protected BALB/c mice from allergic airway inflammation.. Lung CD11b(high)CD103(-)SIRP-alpha+ DC migration is governed by self-CD47 expression, and manipulation of the CD47/SIRP-alpha pathway suppresses CD103(-)SIRP-alpha(+) DC-driven pathogenic T(H)2 responses and airway inflammation.

Topics: Adoptive Transfer; Animals; CD47 Antigen; Cell Movement; Cytokines; Dendritic Cells; Female; Hypersensitivity; Inflammation; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Receptors, Immunologic; Recombinant Fusion Proteins; Th2 Cells

Airway remodeling is a central feature of asthma and includes the formation of new peribronchial blood vessels, which is termed angiogenesis. In a number of disease models, bone marrow-derived endothelial progenitor cells (EPCs) have been shown to contribute to the angiogenic response. In this study we set out to determine whether EPCs were recruited into the lungs in a model of allergic airways disease and to identify the factors regulating EPC trafficking in this model. We observed a significant increase in the number of peribronchial blood vessels at day 24, during the acute inflammatory phase of the model. This angiogenic response was associated with an increase in the quantity of EPCs recoverable from the lung. These EPCs formed colonies after 21 days in culture and were shown to express CD31, von Willebrand factor, and vascular endothelial growth factor (VEGF) receptor 2, but were negative for CD45 and CD14. The influx in EPCs was associated with a significant increase in the proangiogenic factors VEGF-A and the CXCR2 ligands, CXCL1 and CXCL2. However, we show directly that, while the CXCL1 and CXCL2 chemokines can recruit EPCs into the lungs of allergen-sensitized mice, VEGF-A was ineffective in this respect. Further, the blockade of CXCR2 significantly reduced EPC numbers in the lungs after allergen exposure and led to a decrease in the numbers of peribronchial blood vessels after allergen challenge with no effect on inflammation. The data presented here provide in vivo evidence that CXCR2 is critical for both EPC recruitment and the angiogenic response in this model of allergic inflammation of the airways.

Topics: Airway Remodeling; Animals; Antibodies; Cell Movement; Cells, Cultured; Chickens; Endothelial Cells; Female; Hypersensitivity; Mice; Mice, Inbred BALB C; Neovascularization, Pathologic; Ovalbumin; Pneumonia; Receptors, Interleukin-8B; Stem Cells

Specific immunotherapy (SIT) is the only potentially curative treatment for those allergic processes mediated by IgE. We compared the effects of different SITs in mice sensitised with ovalbumin (OVA) Al (OH)(3) : 1) OVA entrapped in particles of poly (D,L-lactic-co-glycolic acid) (PLGA-OVA), 2) Soluble OVA (OVA-sol) and 3) Polymerised OVA (OVA-pol). Serum levels of specific IgE, IgG1, IgG2a and asymmetric IgG, the cutaneous anaphylaxis test (PCA), and the IL-10, IFNgamma and IL-4 levels in culture supernatants of splenocytes challenged with OVA were assessed. Mice treated with PLGA-OVA had higher levels of asymmetric antibodies than non-desensitised mice; a low IgG1 and high IgG2a level was observed together with inhibitory effect in the PCA reaction that reversed in the absence of asymmetric IgG. IL-10 and IFNgamma levels were higher in supernatants from mice treated with PLGA-OVA and OVA-sol than those obtained from non-desensitised controls. Our results suggest that among the different SITs evaluated, PLGA-OVA is the one that best showed an increase in the asymmetric IgG molecules and an effective deviation of the immune response. Furthermore, the increase in the proportion of asymmetric antibodies would be of importance when designing new vaccination strategies for allergy.

Topics: Animals; Cells, Cultured; Chromatography, Affinity; Desensitization, Immunologic; Disease Models, Animal; Glycolates; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-4; Lactic Acid; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Spleen; Time Factors

It is widely established that mast cells (MCs) have a harmful role in asthma, for example by secreting various proinflammatory substances stored within their secretory granule. However, in this study, we show that one of the substances stored within MC granule, chymase, in fact has a protective role in allergic airway inflammation, indicating that MCs may possess both harmful and protective activities in connection with this type of disease. Wild-type (WT) mice and mice lacking mouse MC protease 4 (mMCP-4), a chymase that is functionally homologous to human chymase, were sensitized and challenged with OVA, followed by the assessment of airway physiology and inflammatory parameters. Our results show that the airway hyperresponsiveness was significantly higher in mMCP-4(-/-) as compared with WT mice. Moreover, the degree of lung tissue inflammation was markedly higher in mice lacking mMCP-4 than in WT controls. Histological analysis revealed that OVA sensitization/challenge resulted in a marked increased in the thickness of the smooth muscle cell (SMC) layer and, notably, that the degree of SMC layer thickening was more pronounced in mMCP-4(-/-) animals than in WT controls, thus indicating that chymase may have an effect on airway SMCs. In support of this, mMCP-4-positive MCs were located in the close vicinity of the SMC layer, mainly in the upper airways, and mMCP-4 was shown to be the major chymase expressed in these MCs. Taken together, our results indicate that chymase present in the upper airways protects against allergic airway responses, possibly by regulating SMCs.

Topics: Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Hypersensitivity; Inflammation; Lung; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Myocytes, Smooth Muscle; Ovalbumin; Serine Endopeptidases

Fms-like tyrosine kinase 3 ligand (Flt3L) reverses the features of allergic airway inflammation and increases a Th2-suppressive regulatory lung CD11c(high)CD11b(low) dendritic cell (DC) subset in a mouse model. We examined the migratory pattern and Ag uptake efficiency of lung DC subsets in the therapeutic effect of Flt3L. Lung CD11c(high)CD11b(low) and CD11c(low)CD11b(high) DCs from PBS-treated, OVA-sensitized, and Flt3L-treated/OVA-sensitized BALB/c mice were sorted using MACS and FACS for phenotype analysis. Lymphatic chemokine expression in thoracic lymph nodes was determined by immunohistochemistry. Migration of two lung DC subsets to lymphatic chemokines was examined in vitro using a Transwell chemotaxis assay. Labeled Ag was intranasally delivered into mouse lung to track the migration and Ag uptake of lung DCs. The in vitro cytokine secretion of mediastinal lymph node cells was determined using ELISA. CD11c(low)CD11b(high) DCs have higher expression of CCR5, CCR6, and CCR7, but lower expression of CCR2 than CD11c(high)CD11b(low) DCs. CD11c(low)CD11b(high) DCs in Flt3L-treated/OVA-sensitized mice demonstrated a less mature phenotype, inefficiency in Ag uptake, and impaired migration in vitro to lymphatic chemokine than those in OVA-sensitized mice. Administration of Flt3L decreased the expression of CCR5 and CCR7 in CD11c(low)CD11b(high) DCs in OVA-sensitized mice. Fewer Ag-carrying cells were detected in the lungs and lymph nodes in Flt3L-treated/OVA-sensitized mice than OVA-sensitized mice with a greater decrease in CD11c(low)CD11b(high) DCs. Mediastinal lymph node cells from Flt3L-treated mice secreted higher levels of Th1 cytokines and IL-10 than OVA-sensitized mice in vitro. In conclusion, Flt3L-generated lung immunogenic CD11c(low)CD11b(high) DCs have a less mature phenotype, impaired Ag uptake, and impaired migration to draining lymph nodes.

Topics: Animals; Antigen Presentation; Antigens; Chemotaxis, Leukocyte; Cytokines; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity; Immunohistochemistry; Lymph Nodes; Membrane Proteins; Mice; Mice, Inbred BALB C; Ovalbumin; Phenotype; Pneumonia

We examined cytokine production and allergic reactions in mice fed ad libitum (AL) and subjected to dietary restriction (DR). DR retarded the increase in body weight, and peripheral blood T cells in the DR mice produced less IFN-gamma and more IL-4 in response to immobilized anti-CD3 mAb. Systemic immunization and intranasal challenge with ovalbumin (OVA) induced accumulation of leukocytes into the lung, increase in IL-4 level in bronchoalveolar lavage fluid (BALF), and rise in serum IgE in the AL mice. In contrast, these allergic symptoms were alleviated in the DR mice. Furthermore, the relative proportion of IL-4-producing T cells responsive to OVA was less in the DR mice than the AL mice. DR tended to decrease the proportion and cytolytic activity of NK cells in the spleen, especially in younger mice. These results indicate that DR can prevent the expansion of allergen-specific IL-4-producing T cells followed by suppression of the allergic reaction, but might dampen NK cell activity.

Topics: Animals; Antibody Specificity; Caloric Restriction; Cytokines; Diet; Hypersensitivity; Interleukin-4; Killer Cells, Natural; Male; Mice; Ovalbumin; Spleen; T-Lymphocytes

Lymphocyte stimulation tests (LST) were performed in five dogs sensitised with ovalbumin (OVA) and seven healthy dogs. In addition, all five OVA-sensitised and two control dogs were tested after two in vivo provocations with OVA-containing eye drops. The isolated cells were suspended in culture media containing OVA and were cultured for up to 12 days. Proliferation was measured as reduction in 5,6-carboxylfluorescein diacetate succinimidyl ester (CFSE) intensity by flow cytometry on days 0, 3, 6, 9 and 12. A cell proliferation index (CPI) for each day and the area under the curve (AUC) of the CPI was calculated for each dog. All OVA-sensitised dogs demonstrated increased erythema after conjunctival OVA application. The presence of OVA-specific lymphocytes was demonstrated in 2/5 OVA-sensitised dogs before and 4/5 after in vivo provocation. Using the AUC, the difference between OVA-sensitised and control dogs was significant in all three LST before in vivo provocation (P<0.05) and borderline significant (P=0.053) in 2/3 LST after provocation. The most significant difference in CPI was observed after 9 days of culture (P=0.001). This pilot study indicates that the LST allows detection of rare antigen specific memory T-cells in dogs previously sensitised to, but not concurrently undergoing challenge by a specific antigen.

Topics: Animals; Cell Division; Dog Diseases; Dogs; Erythema; Hypersensitivity; Lymphocyte Activation; Lymphocytes; Ovalbumin; Reference Values

Arachidonate 15-lipoxygenase (LO)-1 has been implicated in allergic inflammation and asthma. The overall effect of 15-LO in allergic inflammation in vivo is, however, unclear. This study investigates systemic allergen sensitization and local allergic airway inflammation and remodeling in mice lacking the murine 12/15-LO, the ortholog to human 15-LO-1. Upon systemic sensitization with intraperitoneal ovalbumin, 12/15-LO-/- mice produced elevated levels of allergen-specific immunoglobulin E compared with wild-type (Wt) controls. However, when challenged with repeated aerosolized allergen, sensitized 12/15-LO-/- mice had an impaired development of airway allergic inflammation compared with Wt controls, as indicated by reduced bronchoalveolar lavage fluid leukocytes (eosinophils, lymphocytes, macrophages) and Th2 cytokines (IL-4, IL-5, IL-13), as well as tissue eosinophils. Allergen-induced airway epithelial proliferation was also significantly attenuated in 12/15-LO-/- mice, whereas goblet cell hyperplasia was unaffected. However, 12/15-LO-/- mice had significantly reduced luminal mucus secretions compared with Wt controls. The repeated allergen challenges resulted in a dramatic increase of alpha-smooth muscle actin-positive alveolar cells in the peripheral airways, a phenomenon that was significantly less developed in 12/15-LO-/- mice. In conclusion, our data suggest that 12/15-LO-/- mice, although having a fully developed systemic sensitization, did not establish a fully developed allergic airway inflammation and associated manifestations of central and peripheral airway remodeling. These data suggest that 12/15-LO-derived metabolites play an important pathophysiologic role in allergen-induced inflammation and remodeling. Hence, pharmacologic targeting of the human 15-LO-1 may represent an attractive therapeutic strategy to control inflammation and remodeling in asthma.

Topics: Allergens; Animals; Antibodies; Antibody Specificity; Apoptosis; Arachidonate 12-Lipoxygenase; Arachidonate 15-Lipoxygenase; Caspase 3; Cell Count; Cytokines; Eosinophilia; Goblet Cells; Hyperplasia; Hypersensitivity; Immunization; Inflammation; Leukocytes; Lung; Mice; Mice, Inbred C57BL; Ovalbumin

Small interfering RNA (siRNA) is a potent means of inducing gene-specific silencing. Gene silencing strategies using siRNA have demonstrated therapeutic benefits in animal models of various diseases, and are currently in clinical trials. However, the utility of gene silencing as a treatment for allergic diseases has not yet been reported. In this study, we report a novel therapy for allergy through gene silencing of CD40, a critical costimulatory molecule and a key factor in allergic immune responses. Silencing CD40 resulted in generation of immunoregulatory dendritic cells (DCs). Administration of CD40 siRNA remarkably reduced nasal allergic symptoms and local eosinophil accumulation in the OVA-induced allergic mice. The OVA-specific T cell response was inhibited after the CD40 siRNA treatment. Additionally, anti-OVA specific IgE and production of IL-4 and IL-5 of T cells stimulated by OVA were significantly decreased in CD40 siRNA-treated mice. Furthermore, we demonstrated that the therapeutic effects by CD40 siRNA were associated with impaired Ag-presenting functions of DCs and B cells, and generation of regulatory T cells. The present study highlights a therapeutic potential of siRNA-based treatment for allergic diseases.

Topics: Animals; B-Lymphocytes; CD40 Antigens; Coculture Techniques; Dendritic Cells; Down-Regulation; Feasibility Studies; Gene Silencing; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunosuppressive Agents; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; RNA, Small Interfering; T-Lymphocytes, Regulatory; Vaccines, Synthetic

Milk consumption from early childhood on has been found to be inversely correlated with allergic sensitization and the onset of bronchial asthma. We tested whether cis-9,trans-11-conjugated linoleic acid (c9,t11-CLA), naturally occurring in milk fat, may prevent allergic sensitization and inhibit airway inflammation in a murine asthma model. BALB/c mice were fed a diet enriched in 1 wt% of c9,t11-CLA or a control diet 7 d prior to and for 32 d during sensitization [d 1 and 14, 100 mg/L ovalbumin (OVA) in adjuvant vs. PBS] and airway challenges (d 28-30, 1% OVA in PBS vs. PBS). Subgroups of mice were coadministered 20 micromol/L of the selective PPARgamma antagonist GW9662 during each OVA challenge. C9,t11-CLA feeding resulted in significantly reduced IgE production and allergen-induced in vivo airway hyperresponsiveness. Further, less mucous plugging of segmental bronchi and significantly reduced interleukin-5 and eosinophils were determined in bronchoalveolar lavage fluids of c9,t11-CLA-fed mice. C9,t11-CLA feeding prevented the downregulation of PPARgamma mRNA in the lung tissues observed after allergen sensitization and airway challenges in control mice. The inhibitory effects of c9,t11-CLA on airway inflammation were partially prevented by coadministration of GW9962. Further, c9,t11-CLA feeding resulted in a significantly lower concentration of the eicosanoid precursor, arachidonic acid, in tissue lipids. These findings demonstrate that dietary c9,t11-CLA can reduce allergic airway inflammation, most likely via a PPARgamma-related mechanism and by reducing eicosanoid precursors. They give new insights into the fatty acid-mediated mechanism of immunomodulation and may represent a step toward an attractive novel strategy in the dietary prevention and treatment of allergic asthma.

Topics: Airway Resistance; Anilides; Animals; Base Sequence; Bronchoalveolar Lavage Fluid; Dietary Fats, Unsaturated; DNA Primers; Female; Hypersensitivity; Inflammation; Linoleic Acids, Conjugated; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; Respiratory System; RNA, Messenger

Diabetic patients are refractory to allergic inflammatory diseases. In this study, the influence of alloxan-induced diabetes on allergic skin inflammation was investigated.. Diabetes was induced by intravenous injection of alloxan into male Wistar rats, and the analyses were performed 21 days later. Animals were actively sensitized with a mixture of aluminium hydroxide plus ovalbumin and challenged intradermally with ovalbumin on day 14.. Diabetic sensitized rats exhibited a less pronounced antigen-induced protein extravasation in the dorsal skin when compared with normal animals. Also, fragments of the dorsal subcutaneous tissue from diabetic sensitized rats showed a reduction in histamine release after stimulation with antigen in vitrowhen compared with fragments obtained from nondiabetic sensitized rats. Optical microscopy analysis revealed that the dorsal skin of diabetic rats showed a marked reduction in dermis thickness, as compared with that seen in normal animals. A significant decrease in the number of skin mast cells was also noted, a phenomenon that paralleled with the reduction in the expression of extracellular matrix components laminin, fibronectin and collagen. Administration of insulin into diabetic rats restored basal mast cell numbers as well as the levels of laminin, fibronectin and collagen.. Our findings show that alloxan diabetes induces downregulation of the skin allergic inflammatory response in rats, and this was correlated with reduction in local mast cell numbers and expression of extracellular matrix components. Lastly, these alterations were reversed with insulin treatment.

Topics: Allergens; Alloxan; Aluminum Hydroxide; Animals; Diabetes Mellitus, Experimental; Down-Regulation; Extracellular Matrix; Histamine Release; Hypersensitivity; Inflammation; Male; Mast Cells; Ovalbumin; Rats; Rats, Wistar; Skin

To determine whether alpha-linked galacto-oligosaccharide (alpha-GOS) prevents allergic peritonitis, BALB/c mice were fed a synthetic diet with and without alpha-GOS supplementation for 7 d, and were then subcutaneously immunized with ovalbumin on days 0 and 7. The mice were challenged by intraperitoneal injection with ovalbumin on day 14, followed by peritoneal lavage on day 15. The total number of peritoneal exudate cells was significantly lower in the mice fed the alpha-GOS diet than in those fed the control diet. Peritoneal lavage fluid from mice fed the alpha-GOS diet not only had less potency to attract peripheral blood leukocytes and peritoneal exudate cells ex vivo, but also had lower concentrations of monocyte chemoattractant protein-1 (MCP-1) and eotaxin. Preincubation of the cells with alpha-GOS failed to affect the migration to peritoneal lavage fluid. We propose that dietary alpha-GOS reduces cell infiltration in allergic peritonitis by reducing antigen-induced elicitation of MCP-1 and eotaxin in mice.

Topics: Animals; Ascitic Fluid; Cell Movement; Chemokine CCL11; Chemokine CCL2; Dietary Supplements; Galactose; Hypersensitivity; Leukocytes; Mice; Mice, Inbred BALB C; Oligosaccharides; Ovalbumin; Peritonitis

Non-selective cation influx through canonical transient receptor potential channels (TRPCs) is thought to be an important event leading to airway inflammation. TRPC6 is highly expressed in the lung, but its role in allergic processes is still poorly understood.. The purpose of this study was to evaluate the role of TRPC6 in airway hyperresponsiveness (AHR) and allergic inflammation of the lung.. Methacholine-induced AHR was assessed by head-out body plethysmography of wild type (WT) and TRPC6(-/-) mice. Experimental airway inflammation was induced by intraperitoneal ovalbumin (OVA) sensitization, followed by OVA aerosol challenges. Allergic inflammation and mucus production were analysed 24 h after the last allergen challenge.. Methacholine-induced AHR and agonist-induced contractility of tracheal rings were increased in TRPC6(-/-) mice compared with WT mice, most probably due to compensatory up-regulation of TRPC3 in airway smooth muscle cells. Most interestingly, when compared with WT mice, TRPC6(-/-) mice exhibited reduced allergic responses after allergen challenge as evidenced by a decrease in airway eosinophilia and blood IgE levels, as well as decreased levels of T-helper type 2 (Th2) cytokines (IL-5, IL-13) in the bronchoalveolar lavage. However, lung mucus production after allergen challenge was not altered by TRPC6 deficiency.. TRPC6 deficiency inhibits specific allergic immune responses, pointing to an important immunological function of this cation channel in Th2 cells, eosinophils, mast cells and B cells.

Topics: Animals; Bronchial Hyperreactivity; Cells, Cultured; Epithelial Cells; Guinea Pigs; Hypersensitivity; Immunoglobulin E; In Vitro Techniques; Interleukin-13; Interleukin-5; Leukocytes; Lung; Mice; Mice, Knockout; Mucus; Muscle Contraction; Muscle, Smooth; Ovalbumin; Pulmonary Eosinophilia; Trachea; TRPC Cation Channels; TRPC6 Cation Channel

Recently, the influence of parasitic infections on the incidence of allergic diseases has become the focus of increased attention. In order to ascertain whether parasite-derived proteins could inhibit the allergic specific Th2 response, we applied excretory-secretory protein (Tl-ES) or total protein (Tl-TP) of the adult worm Toxascaris leonina to asthma model mice prior to or simultaneously with OVA challenge, after which we assessed the OVA-specific Th2 responses. The group subjected to immunization with Tl-ES and Tl-TP (immunized group) evidenced a thinning of the bronchial epithelial and muscle layer, a disruption and shedding of epithelial cells, a reduction in the number of goblet cells, and a reduction in mucus production as compared to the group treated with Tl-ES coupled with OVA challenge (challenge with OVA groups) and the OVA-induced asthma group. The administration of Tl-ES and Tl-TP, regardless of injection time, was shown to inhibit the recruitment of inflammatory cells into the airway, and in particular, macrophages, neutrophils, and lymphocytes were significantly reduced as the result of the parasite proteins. However, the total number of eosinophils was slightly reduced as the result of the administration of parasite proteins. Sensitization and OVA challenge was shown to accelerate the secretion of Th2 cytokines (IL-4 and IL-5) within the lung, but in the immunized groups, those levels were lower. The administration of Tl-TP and OVA challenge group also evidenced a significant reduction in IL-4 levels as compared to the OVA-challenged group. The concentrations of Th2 cytokines in the Tl-ES and OVA challenge group were more similar to those observed in the OVA-challenged group. The concentration of IL-10 and TGF-beta in the lung was decreased substantially in the OVA-only challenge group, but the Tl-TP immunized group exhibited significantly induced IL-10 cytokine. OVA-specific IgG2a, IgG1, and IgE levels in the immunized groups were significantly lower than those detected in the OVA-challenged group. In conclusion, parasite-derived protein is able to inhibit OVA-specific Th2 responses, and in particular, immunization with parasite proteins exerts a more profound protective effect than is seen with the treatment of allergic reactions. The results of our study are encouraging in terms of our further understanding of the molecular basis of immune evasion by nematodes.

Topics: Animals; Asthma; Enzyme-Linked Immunosorbent Assay; Female; Helminth Proteins; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells; Toxascariasis; Toxascaris

Schinus is a genus of the Anacardiaceae family and contains Schinus terebinthifolius, the Brazilian pepper tree that is widely used in folk medicine. We investigate the anti-allergic activity of the ethyl acetate fraction of S. terebinthifolius Raddi (ST fraction). HPLC analysis reveled that gallic acid, methyl gallate and 1,2,3,4,6-pentagalloylglucose are the major aromatic components of the fraction. Oral pre-treatment with the ST fraction (100 mg/kg) significantly inhibited paw edema induced by compound 48/80 (100 ng/paw) and to a lesser extent, the allergic paw edema (OVA, 3 microg/paw). The ST fraction (100 and 200 mg/kg) also inhibited the edema induced by histamine (100 microg/paw), preventing mast cell degranulation and, consequently, histamine release in Wistar rat peritoneal mast cells induced by C 48/80 (5 microg/mL). This histamine inhibition was also observed after mast cell pre-treatment with both methyl gallate and 1,2,3,4,6-pentagalloylglucose (100 microg/mL), the isolated compounds from the ethyl acetate fraction. Pre-treatment with the ST fraction (100 mg/kg) significantly inhibited total leukocyte and eosinophil accumulation in pleural cavities 24 h after the intrathoracic injection of OVA (12.5 microg/cavity). This effect was related to the inhibition of CCL11/eotaxin and CCL5/RANTES in pleural lavage fluid. Pre-treatment with this fraction (100 mg/kg) failed to reduce the cell influx that was observed after LPS-injection into pleural cavity (250 ng/cavity). These findings demonstrate the anti-allergic effect of the ST fraction, which includes the inhibition of edema formation and histamine release caused by mast cell degranulation and eosinophil influx into the pleural cavity probably reflected by the decreased levels of chemokines in recovered pleural lavage fluid.

Topics: Anacardiaceae; Animals; Anti-Allergic Agents; Chemokines; Edema; Histamine; Histamine Release; Hypersensitivity; Immunoglobulin E; Lipopolysaccharides; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Leaves; Pleurisy; Promethazine; Rats; Rats, Wistar

The addition of a nitric oxide (NO)-releasing moiety to prednisolone was shown to enhance the anti-inflammatory activity of this glucocorticoid in some experimental conditions, but its effectiveness in the context of eosinophilic inflammation remains to be elucidated.. This study compared the anti-inflammatory effect of prednisolone to a NO-releasing derivative of prednisolone, NCX-1015, using a model of allergen-evoked eosinophil recruitment in rats. The efficacy of a NO-donor compound, DETA-NONOate, was also assessed for comparison.. Wistar rats were actively sensitized with Al(OH)(3) plus ovalbumin and 14 days later challenged with antigen intrapleurally. Treatments were performed locally 1 h before challenge. Cysteinyl-leucotrienes (Cys-LT) and eotaxin were measured by ELISA.. Antigen challenge induced an eosinophil infiltration at 12 h, maximal at 24 h. It also caused an increase in the levels of Cys-LTs in the pleural exudate and in the expression of 5-lipoxygenase (5-LO) in infiltrated leucocytes at 6 h, peaking at 12 h and persisting for at least 24 h. Treatment with equimolar doses of prednisolone and NCX-1015 inhibited the late eosinophil infiltration, although the dose required to produce maximal inhibition was about one-tenth that of prednisolone. Cys-LT generation and 5-LO expression were inhibited by NCX-1015 but not by prednisolone. Treatment with prednisolone combined with the NO-donor DETA-NONOate led to a greater inhibition of the eosinophilia and Cys-LT generation as compared with either drug alone. Administration of the steroid receptor antagonist RU 486, 1 h before prednisolone and NCX-1015, abolished the inhibitory effect of the former, under conditions where it only partially affected the latter.. Our findings indicate that NCX-1015 provided a greater anti-inflammatory effect than prednisolone on the allergic eosinophil recruitment in rats, suggesting that NO-releasing steroids can be considered as a promising therapeutic approach to allergic diseases.

Topics: Animals; Anti-Inflammatory Agents; Arachidonate 5-Lipoxygenase; Chemokine CCL11; Cysteine; Disease Models, Animal; Drug Therapy, Combination; Eosinophilia; Eosinophils; Hypersensitivity; Leukocytes; Leukocytes, Mononuclear; Leukotrienes; Male; Mifepristone; Neutrophils; Nitric Oxide Donors; Nitroso Compounds; Ovalbumin; Pleural Cavity; Pleurisy; Prednisolone; Rats; Rats, Wistar; Receptors, Glucocorticoid

Thymic stromal lymphopoietin (TSLP) is a cytokine expressed by epithelial cells, including keratinocytes, and is important in allergic inflammation. Allergic skin inflammation elicited by epicutaneous immunization of mice with ovalbumin (OVA), a potential model of atopic dermatitis, was severely impaired in TSLPR(-/-) mice, as evidenced by decreased infiltration of eosinophils and decreased local expression of T helper 2 (Th2) cytokines. However, secretion of Th2 cytokines by splenocytes from epicutaneous sensitized TSLPR(-/-) mice in response to OVA was normal. Skin dendritic cells from TSLPR(-/-) mice were normal in their ability to migrate to draining lymph nodes, express activation markers, and induce proliferation and Th2 cytokine production by naïve T cells. CD4(+) T cells from TSLPR(-/-) mice expressed the skin homing receptor E-selectin ligand normally, and homed to the skin normally, but failed to transfer allergic skin inflammation to WT recipients. TSLP enhanced Th2 cytokine secretion in vitro by targeting TSLPR on antigen specific T cells. Intradermal injection of anti-TSLP blocked the development of allergic skin inflammation after cutaneous antigen challenge of OVA immunized WT mice. These findings suggest that TSLP is essential for antigen driven Th2 cytokine secretion by skin infiltrating effector T cells and could be a therapeutic target in allergic skin inflammation.

Topics: Animals; Cell Movement; Cell Proliferation; Cells, Cultured; Cytokines; Dendritic Cells; Dermatitis; Female; Hypersensitivity; Immunity, Innate; Immunoglobulins; Lymph Nodes; Mice; Mice, Knockout; Ovalbumin; Receptors, Cytokine; Spleen; T-Lymphocytes; Thymic Stromal Lymphopoietin

Recent evidence suggests that asthma leads to inflammation and remodeling not only in the airways but also in pulmonary vessels and parenchyma. In addition, some studies demonstrated that aerobic training decreases chronic allergic inflammation in the airways; however, its effects on the pulmonary vessels and parenchyma have not been previously evaluated. Our objective was to test the hypothesis that aerobic conditioning reduces inflammation and remodeling in pulmonary vessels and parenchyma in a model of chronic allergic lung inflammation. Balb/c mice were sensitized at days 0, 14, 28, and 42 and challenged with ovalbumin (OVA) from day 21 to day 50. Aerobic training started on day 21 and continued until day 50. Pulmonary vessel and parenchyma inflammation and remodeling were evaluated by quantitative analysis of eosinophils and mononuclear cells and by collagen and elastin contents and smooth muscle thickness. Immunohistochemistry was performed to quantify the density of positive cells to interleukin (IL)-2, IL-4, IL-5, interferon-gamma, IL-10, monocyte chemotatic protein (MCP)-1, nuclear factor (NF)-kappaB p65, and insulin-like growth factor (IGF)-I. OVA exposure induced pulmonary blood vessels and parenchyma inflammation as well as increased expression of IL-4, IL-5, MCP-1, NF-kappaB p65, and IGF-I by inflammatory cells were reduced by aerobic conditioning. OVA exposure also induced an increase in smooth muscle thickness and elastic and collagen contents in pulmonary vessels, which were reduced by aerobic conditioning. Aerobic conditioning increased the expression of IL-10 in sensitized mice. We conclude that aerobic conditioning decreases pulmonary vascular and parenchymal inflammation and remodeling in this experimental model of chronic allergic lung inflammation in mice.

Topics: Anaerobiosis; Animals; Conditioning, Psychological; Exercise Test; Hypersensitivity; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Pulmonary Circulation; Reference Values; Time Factors

Currently, no treatment is available for food allergy and strict avoidance of the allergenic food remains the only way to manage the allergy. New strategies leading to a safe and efficacious food allergy treatment are required. Modified vaccinia virus Ankara (MVA), which allows high levels of expression of recombinant protein in vivo and gives rise to a Th1-biased specific immune response, was used as a prophylactic vaccine in a murine model of ovalbumin (OVA) allergy.. An MVA-OVA vector vaccine was prepared. Female BALB/c mice were vaccinated twice with a MVA-OVA vector vaccine, followed by sensitization with OVA plus alum. OVA-specific immunoglobulin E(IgE) activity was measured by mediator release from rat basophilic leukaemia cells, whereas specific IgG subclass titers were determined by enzyme-linked immunosorbent assay.. Expression of immunological active OVA in mammalian cells was demonstrated. OVA-specific IgE levels in sera from MVA-OVA-vaccinated mice were reduced and appeared delayed. The vaccine-mediated immune modulation was dose-dependent; the highest vaccine dose protected 50% of the animals from allergic sensitization. Upon sensitization, similar OVA-specific IgG1 titers were found in all mice, but the OVA-specific IgG2a antibody levels were strongly increased in MVA-OVA-vaccinated mice, signifying a Th1-biased and, non-allergic immune response.. Prophylactic vaccination with MVA-OVA delays and in part even prevents the onset of a successful allergen-specific sensitization. Recombinant MVA, which fulfills the requirements for clinical application, is a promising candidate vector for the development of novel approaches to allergen-specific prophylactic vaccination and specific immunotherapy.

Topics: Animals; Female; Hypersensitivity; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Th1 Cells; Vaccines; Vaccinia virus

Asthma is characterized as a chronic inflammatory disorder of the airways. Phosphodiesterases (PDE), which hydrolyze cAMP, are considered to play important roles in asthma. We previously reported that acetamide-45 could inhibit cAMP-PDE activity, and histamine- and methacholine-induced contractions of isolated guinea pig trachea. The purpose of this study is to determine whether this agent could suppress allergic-induced airway hyperresponsiveness (AHR) and airway inflammation in allergic mice.. A mouse model for asthma was used to investigate acetamide-45 on the airway lesions compared with glucocorticoids. The study was conducted on mice sensitized and challenged with ovalbumin and the whole body plethysmography was carried out to assess AHR. The bronchoalveolar lavage (BAL) histopathology was examined.. We found that acetamide-45 significantly inhibited the enhanced hyperresponsiveness and eosinophil recruitment in airways with elimination of cAMP-PDE activity in lung tissue. Elevated IL-4 and IL-5 in bronchoalveolar lavage fluid (BALF) in asthmatic mice were markedly decreased.. Our results indicate that the agent has a potential role in inflammatory disease.

Topics: Acetamides; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Eosinophils; Hypersensitivity; Indoles; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-5; Leukocytes; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphodiesterase Inhibitors

To investigate the allergic reaction in neonatal streptozotocin (nSTZ)-induced diabetes mellitus.. Male newborn Wistar rats were made diabetic by the injection of streptozotocin (160 mg/kg, i. p.) and used 8 weeks thereafter.. Animals were sensitized against ovalbumin (OA, 50 microg and Al(OH)3, 5 mg, s. c.) and challenged 14 or 21 days thereafter.. OA-induced airway inflammation and OA-induced pleurisy models were used to investigate leukocyte migration (total and differential leukocyte counts) and lung vascular permeability (Evans blue dye extravasation).. nSTZ-diabetic rats presented glucose intolerance and insulin resistance. Relative to controls, nSTZ rats exhibited a 30% to 50% reduction in lung vascular permeability. Leukocyte infiltration in both models of allergen-induced inflammation, and number of pleural mast cells did not differ between groups.. Data suggest that the reduction of allergic inflammatory reactions in nSTZ rats is restricted to microvascular dysfunctions and associated, probably, with insulin resistance in lung microvascular endothelium.

Topics: Animals; Animals, Newborn; Capillary Permeability; Diabetes Mellitus, Experimental; Glucose Tolerance Test; Hypersensitivity; Inflammation; Insulin Resistance; Male; Ovalbumin; Pleurisy; Rats; Rats, Wistar; Streptozocin

Corticosteroids are known to inhibit bronchial hyperresponsiveness (BHR) and allergic inflammation but there is little information on its dose-dependence. We examined the effect of different doses of the glucocorticosteroid budesonide in an allergic model. Brown-Norway rats were sensitised to ovalbumin (OVA) and pretreated with an intra-gastric dose of budesonide (0.1, 1.0, or 10 mgkg(-1)). Exposure to OVA induced BHR, accumulation of eosinophils in the bronchoalveolar lavage (BAL) fluid and in the airways submucosa. Budesonide dose-dependently inhibited BAL fluid influx of lymphocytes, eosinophils and neutrophils, tissue eosinophils and lymphocytes and BHR. At 0.1 mgkg(-1), budesonide did not inhibit these parameters but at 1 mgkg(-1), BAL fluid eosinophils and T-cells, and submucosal T-cells were significantly reduced. At 10 mgkg(-1), budesonide suppressed BHR, BAL fluid inflammatory cells numbers and tissue eosinophilia. T-cell numbers were more related to BHR than eosinophil numbers. Budesonide inhibited both airway inflammation and BHR, but BAL fluid eosinophil cell counts may be dissociated from BHR.

Topics: Acetylcholine; Animals; Blood Cell Count; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Budesonide; Disease Models, Animal; Dose-Response Relationship, Drug; Glucocorticoids; Hypersensitivity; Male; Ovalbumin; Rats

Peroxisome proliferator-activated receptor-alpha (PPARalpha) is implicated in the control of airway inflammation. However, little is known so far about PPARalpha expression and regulation in the lung. Our aim was to assess PPARalpha expression in the lung from normal mice, as well as to investigate its regulation during airway inflammation or in response to anti-inflammatory agents. The PPARalpha activator, fenofibrate, the glucocorticoid, dexamethasone or vehicle was administered to normal mice, to mice exposed to tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) or to ovalbumin (OVA)-sensitized and challenged animals. PPARalpha expression was assessed by quantifying PPARalpha mRNA levels using real-time quantitative PCR after reverse-transcription of total lung RNA. Airway inflammation was evaluated by determining total and differential cell counts, as well as TNF-alpha production in bronchoalveolar lavage fluids. PPARalpha mRNA was found at significant levels in the lung from normal mice. This expression was increased by 65% (p<0.05) and 55% (p<0.05) in animals treated with fenofibrate and dexamethasone, respectively. In mice exposed to TNF-alpha or LPS, as well as in animals sensitized and challenged with OVA, that exhibited airway inflammation, PPARalpha mRNA was decreased by 60% (p<0.05), 43% (p<0.05) and 50% (p<0.05), respectively. In mice exposed to LPS, down-regulation of PPARalpha was maximal at 4h, whereas TNF-alpha production and cell infiltration peaked at 2 and 24h, respectively. In the lung of mice exposed to LPS or OVA and treated with fenofibrate or dexamethasone, PPARalpha down-regulation was suppressed, while airway inflammation was abolished. Our data showed that PPARalpha is constitutively expressed in mouse lung and down-regulated in response to TNF-alpha or upon acute or allergic airway inflammation. Fenofibrate and dexamethasone upregulated PPARalpha in normal lung and suppressed PPARalpha down-regulation associated with airway inflammation. Taken together, our data show that PPARalpha expression is inversely regulated with lung inflammation.

Topics: Animals; Anti-Inflammatory Agents; Bronchoalveolar Lavage Fluid; Dexamethasone; Down-Regulation; Fenofibrate; Glucocorticoids; Hypersensitivity; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred C57BL; Ovalbumin; PPAR alpha; Respiratory Distress Syndrome; RNA, Messenger; Transcription Factors; Tumor Necrosis Factor-alpha

Increased resistance in the small airways is a major contributor of airway obstruction in asthma. The role of leukotrienes (LT) in determining inflammation and obstruction of small size bronchi is not completely understood. Here, we have examined the effect of the cysteinyl-leukotriene (CysLT 1) receptor antagonist, montelukast, against the bronchoconstriction and inflammatory responses induced by exogenous leukotriene and by allergen challenge in small size (

Topics: Acetates; Animals; Bronchi; Bronchoconstriction; Capillary Permeability; Cyclopropanes; Eosinophils; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Leukotriene Antagonists; Male; Microcirculation; Ovalbumin; Quinolines; Receptors, Leukotriene; Sulfides

The prevalence and morbidity of asthma, a chronic inflammatory airway disease, is increasing. Animal models provide a meaningful but limited view of the mechanisms of asthma in humans. A systems-level view of asthma that integrates multiple levels of molecular and functional information is needed. For this, we compiled a gene expression compendium from five publicly available mouse microarray datasets and a gene knowledge base of 4,305 gene annotation sets. Using this collection we generated a high-level map of the functional themes that characterize animal models of asthma, dominated by innate and adaptive immune response. We used Module Networks analysis to identify co-regulated gene modules. The resulting modules reflect four distinct responses to treatment, including early response, general induction, repression, and IL-13-dependent response. One module with a persistent induction in response to treatment is mainly composed of genes with suggested roles in asthma, suggesting a similar role for other module members. Analysis of IL-13-dependent response using protein interaction networks highlights a role for TGF-beta1 as a key regulator of asthma. Our analysis demonstrates the discovery potential of systems-level approaches and provides a framework for applying such approaches to asthma.

Topics: Algorithms; Allergens; Animals; Asthma; Disease Models, Animal; Gene Expression Profiling; Humans; Hypersensitivity; Immunity, Innate; Interleukin-13; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Knockout; Models, Biological; Oligonucleotide Array Sequence Analysis; Ovalbumin; Protein Interaction Mapping; Reproducibility of Results; Systems Biology; Transcription, Genetic; Transforming Growth Factor beta1

Traditional therapies for asthma and allergic rhinitis (AR) such as corticosteroids and antihistamines are not without limitations and side effects. The use of complementary and alternative approaches to treat allergic airways disease, including the use of herbal and dietary supplements, is increasing but their efficacy and safety are relatively understudied. Previously, we have demonstrated that gamma-tocopherol (gammaT), the primary form of dietary vitamin E, is more effective than alpha-tocopherol, the primary form found in supplements and tissue, in reducing systemic inflammation induced by non-immunogenic stimuli.. We used allergic Brown Norway rats to test the hypothesis that a dietary supplement with gammaT would protect from adverse nasal and pulmonary responses to airway allergen provocation.. Ovalbumin (OVA)-sensitized Brown Norway rats were treated orally with gammaT before intranasal provocation with OVA. Twenty-four hours after two challenges, histopathological changes in the nose, sinus and pulmonary airways were compared with gene expression and cytokine production in bronchoalveolar lavage fluid and plasma.. We found that acute dosing for 4 days with gammaT was sufficient to provide broad protection from inflammatory cell recruitment and epithelial cell alterations induced by allergen challenge. Eosinophil infiltration into airspaces and tissues of the lung, nose, sinus and nasolacrimal duct was blocked in allergic rats treated with gammaT. Pulmonary production of soluble mediators PGE(2), LTB(4) and cysteinyl leukotrienes, and nasal expression of IL-4, -5, -13 and IFN-gamma were also inhibited by gammaT. Mucous cell metaplasia, the increase in the number of goblet cells and amounts of intraepithelial mucus storage, was induced by allergen in both pulmonary and nasal airways and decreased by treatment with gammaT.. Acute treatment with gammaT inhibits important inflammatory pathways that underlie the pathogenesis of both AR and asthma. Supplementation with gammaT may be a novel complementary therapy for allergic airways disease.

Topics: Animals; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dietary Supplements; Eosinophilia; gamma-Tocopherol; Gene Expression; Hyperplasia; Hypersensitivity; Lung; Male; Nasal Mucosa; Ovalbumin; Paranasal Sinuses; Rats; Rats, Inbred BN; Respiratory Mucosa; Respiratory Tract Diseases; Rhinitis

Dietary fatty acids have been shown to influence allergic sensitisation. Both n-3 and n-6 PUFA are involved in targeted mediation of inflammatory responses during allergic sensitisation and manifestation of atopic diseases. In the present experiments we investigated whether supplementation of DHA-enriched fish oil partly substituting dietary sunflower-seed oil, in comparison with sunflower-seed oil, supplemented to mice influences fatty acid composition of serum lipid classes. The effects of the two different diets were also investigated depending on allergic sensitisation. Supplementation of DHA and EPA in doses of 2 and 0.12 % (w/w) to non-sensitised and sensitised mice resulted in significantly increased percentile contributions of DHA to all lipid classes. In contrast, serum values of the n-6 PUFA arachidonic acid (AA) were significantly lower, both in non-sensitised and sensitised mice fed the DHA-enriched diet. The fatty acid composition of serum lipids also reflected allergic sensitisation: the EPA:AA ratio in TAG, cholesteryl esters and phospholipids in non-supplemented animals fell to 23, 29 and 29 % respectively of the original value after allergic sensitisation, whereas it decreased to 70, 80 and 76 % respectively only in the animals supplemented with DHA. In summary, allergic sensitisation alone decreased significantly the EPA:AA ratios in serum TAG, while concomitant supplementation of DHA-enriched fish oil ameliorated this decrease. We postulate from the present results that the amelioration of the severity of allergic sensitisation after DHA supplementation may be linked to altered ratios of the eicosanoid precursors EPA and AA as well as DHA needed for further metabolic activation to pro- or anti-inflammatory bioactive lipids.

Topics: Animals; Arachidonic Acid; Cholesterol Esters; Dietary Supplements; Docosahexaenoic Acids; Eicosapentaenoic Acid; Enzyme-Linked Immunosorbent Assay; Fatty Acids; Fatty Acids, Omega-3; Female; Fish Oils; Galactosylceramides; gamma-Linolenic Acid; Hypersensitivity; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Phospholipids; Plant Oils; Sunflower Oil; Triglycerides

Live BCG administered intranasally to mice inhibits the development of ovalbumin (OVA)-induced eosinophilia and airway hyperresponsiveness (AHR). It is unacceptable to treat human subjects intranasally with live BCG.. We investigated whether BCG killed by extended freeze-drying (EFD) and subcutaneously injected has a protective effect in murine and guinea pig models of allergic airway inflammation.. Mice were OVA sensitized (days 0 and 7), treated subcutaneously (day 14) with EFD and live or heat-killed BCG, and then OVA challenged (day 42). OVA-sensitized mice (days 0 and 7) were challenged (day 14) and EFD treated (day 18) before OVA rechallenge (day 46) to demonstrate the capacity of EFD to reverse the established lung inflammation. Guinea pigs were OVA sensitized (days 0 and 14), treated intradermally (day 35) with EFD, and OVA challenged (days 90-105).. In mice and guinea pigs EFD treatment reduced AHR. Among 3 BCG preparations, only EFD efficiently reduced AHR, eosinophilia, and the recruitment of dendritic cells to the lungs after OVA challenge. The protective effect of EFD is associated with production of the immunoregulatory cytokine IL-10. Moreover, EFD treatment did not induce toxic effects or delayed-type hypersensitivity to mycobacterial antigens; that is, it did not interfere with the diagnosis of tuberculosis.. EFD administered subcutaneously inhibits the development of allergic airway inflammation and prevents AHR without inducing delayed-type hypersensitivity and side effects associated with live or heat-killed BCG.

Topics: Animals; BCG Vaccine; Bronchial Hyperreactivity; Bronchitis; Dendritic Cells; Eosinophilia; Freeze Drying; Guinea Pigs; Hypersensitivity; Injections, Subcutaneous; Interleukin-10; Lung; Male; Mice; Ovalbumin; Pneumonia; Vaccines, Inactivated

In the present study, we show that the intra-thoracic injection of ovalbumin (OVA, 12.5 microg per cavity) into C57BL/10 mice induced a significant increase in gammadelta T lymphocyte numbers in the pleural cavity, blood and thoracic lymph node of challenged mice. Such increase was significant within 12 h, peaked within 48 h and returned to basal counts within 120 h. Levels of CC chemokine ligand (CCL)-2/monocyte chemotactic protein-1, CCL5/regulated upon activation, normal T cell expressed and secreted, CCL3/macrophage inflammatory protein-1 alpha and CCL25/thymus-expressed chemokine were above control values in pleural washes recovered 24 h after OVA challenge (OPW) and were likely produced by pleural macrophages and mesothelial cells. Antigenic challenge also induced an up-regulation in CC chemokine receptor (CCR)-2, CCR5 and CCR9 on gammadelta T cells from pleural cavities, blood and lymph nodes, suggesting that cells found in mice pleural cavity migrate from secondary lymphoid organs into the inflammatory site via blood stream. The in vitro neutralization of CCL2 (but not of CCL3, CCL5 or CCL25) abrogated OPW-induced gammadelta T lymphocyte transmigration. Confirming such results, the in vivo administration of alpha-CCL2 mAb inhibited gammadelta T lymphocyte accumulation in the pleural cavity of challenged mice, whereas the blockade of CCL3, CCL5 or CCL25 showed no effect on gammadelta T cell mobilization. In addition, OVA challenge failed to induce gammadelta T lymphocyte accumulation in the pleural cavity of C57BL/6 CCR2 knockout mice, which also showed decreased numbers of these cells in blood and lymph nodes when compared with wild-type mice. Overall, such results demonstrate that CCR2/CCL2 pathway is crucial for gammadelta T lymphocyte mobilization during the allergic response.

Topics: Animals; Cells, Cultured; Chemokine CCL2; Chemotaxis, Leukocyte; Hypersensitivity; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pleura; Pleurisy; Receptors, Antigen, T-Cell, gamma-delta; Receptors, CCR2; T-Lymphocytes

Modulation of leukocyte recruitment through intervention with chemokine receptors is an attractive, therapeutic strategy. Recently, we have shown that n-Nonanoyl (NNY)-CCL14 internalizes and desensitizes human (h)CCR3, resulting in the inactivation of eosinophils. In this study, we investigated the interaction of NNY-CCL14 with CCR1 and CCR5 and the relevance of these NNY-CCL14 receptors on its in vivo effects in allergic airway inflammation. NNY-CCL14 has inactivating properties on CCR1(+) and CCR5(+) cell lines and primary leukocytes. It desensitizes hCCR1- and hCCR5-mediated calcium release and internalizes these receptors from the cellular surface. Treatment of OVA-sensitized BALB/c mice with NNY-CCL14 resulted in reduced pulmonary inflammation. Above all, it is demonstrated that systemic treatment with NNY-CCL14 down-modulates CCR5 from the surface of lymphocytes in vivo. Although NNY-CCL14 acts on murine lymphocytes and internalizes CCR5, it does not internalize CCR3 on mouse eosinophils, showing species selectivity regarding this particular receptor. Therefore, the inhibitory effects of NNY-CCL14 in murine models of allergic airway inflammation can be assigned to its interaction with CCR5. The presented results substantiate the relevance of CCR5 as a target for allergic airway inflammation.

Topics: Animals; CCR5 Receptor Antagonists; Chemokine CCL11; Chemokine CCL3; Chemokines, CC; Humans; Hypersensitivity; Inflammation; Mice; Ovalbumin; Receptors, CCR1; Receptors, CCR5; T-Lymphocytes

The relative contributions of the allergen-specific early-phase nasal response and nonspecific nasal response and mast cells to the pathophysiology of allergic rhinitis are not well defined.. To determine the contributions of specific reactivity, nonspecific reactivity, and mast cells to the development of early-phase and late-phase responses using a mouse model of allergic rhinitis.. Sensitized wild-type and FcvarepsilonRI-deficient (FcvarepsilonRI-/-) mice were exposed to allergen for 3, 5, or 12 days. As indicators of nasal reactivity, respiratory frequency and nasal resistance were monitored.. Sensitized mice exposed to 3 days of nasal allergen challenge showed a nonspecific early-phase response. As the number of allergen exposures increased, there was progressive diminution in nonspecific responses with increased allergen-specific early-phase responses and a late-phase response. Sensitized FcvarepsilonRI-/- mice did not develop nonspecific nasal responses or late-phase responses, but transfer of in vitro-differentiated wild-type mast cells into FcvarepsilonRI-/- mice restored nonspecific early-phase nasal responses but not the late-phase response.. These data identify the nonspecific nasal response as a major contributor to the early-phase response, especially during initial allergen exposure, and is dependent on mast cells. Increasing allergen exposure results in increasing allergen-specific responses, converting the nonspecific early-phase response to a late-phase response that is allergen-specific and mast cell-independent.

Topics: Allergens; Ambrosia; Animals; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Nasal Provocation Tests; Ovalbumin; Rhinitis; Time

Racemic albuterol is an equimolar mixture of two isomers, (R) and (S). Whether (R) and (S) isomers and the combination of both exert different effects in immune activation is not well defined. We analyzed the effects of (R+S)-albuterol, (R)-albuterol and (S)-albuterol in a murine model of allergic pulmonary inflammation and in activated T cells. Mice (C57BL/6) sensitized and aerosol challenged with the allergen ovalbumin (OVA) or phosphate buffered saline (PBS) were treated with (R)-albuterol, (S)-albuterol or (R+S)-albuterol. Following administration of (R)-albuterol, allergen induced bronchoalveolar lavage eosinophils and IgE showed a decrease, albeit not significantly by ANOVA. As T cells are important in allergic inflammation, we asked whether (R+S), (R) or (S)-albuterol might differ in effects on T cells and on the activity of the inflammatory transcription factor NF-kappaB. In activated T cells, (R)-albuterol administration decreased levels of inflammatory cytokines and NF-kappaB activity. These studies suggest that (R)-albuterol decreases cytokine secretion and NF-kappaB activity in T cells.

Topics: Adrenergic beta-Agonists; Albuterol; Animals; Cell Line; Cells, Cultured; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung Diseases; Mice; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Pneumonia; Protein Isoforms; Spleen; T-Lymphocytes

We already demonstrated that adoptive transfer of alveolar macrophages (AMs) from non-allergic rats into AM-depleted allergic rats prevents airway hyperresponsiveness (AHR). We also showed that AMs from non-sensitized, but not from sensitized, allergy-prone rats can prevent AHR following allergen challenge in sensitized allergic animals, establishing the importance of rat immunological status on the modulation of AM functions and suggesting that an allergic lung environment alters AM functions.. We investigated how the activation of allergic AMs can be modulated to reinstitute them with their capacity to reduce AHR.. AMs from sensitized Brown Norway rats were cultured ex vivo for up to 18 h in culture media to deprogram them from the influence of the allergic lung before being reintroduced into the lung of AM-depleted sensitized recipient. AHR and cytokines in bronchoalveolar lavage (BAL) were measured following allergen challenge. AMs stimulated ex vivo with Bacillus Calmette-Guerin (BCG) were used as positive controls as BCG induces a T-helper type 1 activation in AMs.. AMs ex vivo cultured for 4-18 h reduced AHR to normal level. Interestingly, pro-allergic functions of AMs were dampened by 18 h culture and they reduced AHR even after spending 48 h in an allergic lung microenvironment. Furthermore, transfer of cultured AMs caused an increase in the levels of IFN-gamma and IL-12 in BAL when compared with their ovalbumin control. After 18 h of ex vivo culture, AMs expressed reduced levels of TNF, IL-1alpha, IL-6, and Arginase-2 mRNAs compared with freshly isolated AMs, suggesting that ex vivo culture exempted AMs from lung stimuli that affected their functions.. There is a significant crosstalk between lung microenvironment and AMs, affecting their functions. It is also the first report showing that sensitized AMs can be modulated ex vivo to reduce lung pro-allergic environment, opening the way to therapies targetting AMs.

Topics: Animals; Arginase; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Separation; Cells, Cultured; Cytokines; Hypersensitivity; Liposomes; Macrophages, Alveolar; Ovalbumin; Rats; Rats, Inbred BN; RNA, Messenger; Time Factors

Allergic asthma is a chronic disease characterized by airway obstruction in response to allergen exposure. It results from an inappropriate T helper type 2 response to environmental airborne antigens and affects 300 million individuals. Its prevalence has increased markedly in recent decades, most probably as a result of changes in environmental factors. Exposure to environmental antigens during infancy is crucial to the development of asthma. Epidemiological studies on the relationship between breastfeeding and allergic diseases have reached conflicting results. Here, we have investigated whether the exposure of lactating mice to an airborne allergen affects asthma development in progeny. We found that airborne antigens were efficiently transferred from the mother to the neonate through milk and that tolerance induction did not require the transfer of immunoglobulins. Breastfeeding-induced tolerance relied on the presence of transforming growth factor (TGF)-beta during lactation, was mediated by regulatory CD4+ T lymphocytes and depended on TGF-beta signaling in T cells. In conclusion, breast milk-mediated transfer of an antigen to the neonate resulted in oral tolerance induction leading to antigen-specific protection from allergic airway disease. This study may pave the way for the design of new strategies to prevent the development of allergic diseases.

Topics: Animals; Antigens; Asthma; Bronchial Hyperreactivity; Female; Hypersensitivity; Immune Tolerance; Immunoglobulins; Lactation; Maternal Exposure; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Milk; Ovalbumin; Pneumonia; T-Lymphocytes, Regulatory

Evidence indicates that environment pollutants from fossil fuel combustion compromise the immune system by enhancing allergic reactions and damaging the respiratory tract. This study was performed to investigate the effects of motorcycle exhaust particles (MEP), a major air pollutant especially in the urban areas of Taiwan, on allergen-induced airway inflammatory reactions in lab animals. BALB/c mice were intratracheally instilled with ovalbumin (OVA), MEP, or phosphate-buffered saline, 3 times every 2 wk. Airway hyperresponsiveness was measured in unrestrained mice by barometric plethsmography. Bronchoalveolar lavage fluid (BALF) and serum from treated animals were collected for cytokine and antibody determination by enzyme-linked immunosorbent assay (ELISA). Lung tissue stained with hematoxylin/eosin was examined. Data showed that MEP augmented OVA-induced airway inflammation; characterized by infiltration of eosinophils and neutrophils in BALF and lung tissue inflammation. The combination of OVA and MEP markedly increased interleukin-4 (IL-4), interleukin-5 (IL-5), and tumor necrosis factor-alpha (TNF-alpha) protein levels in BALF. In addition, MEP also augmented OVA-induced rise in OVA-specific immunoglobulin (Ig) G1 and IgE and airway hyperresponsiveness. Pretreated lavage cells with mitogen-activated protein kinase (MAPK) inhibitors showed that TNF-alpha release was significantly inhibited. This study found that MEP augmented antigen-induced allergic airway inflammation and airway hyperresponsiveness through a Th2-dominant pathway.

Topics: Air Pollutants; Animals; Antigens; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Mice; Mice, Inbred BALB C; Motorcycles; Ovalbumin; Respiratory System; Vehicle Emissions

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Bronchoconstrictor Agents; Disease Models, Animal; Drug Synergism; Hypersensitivity; Ovalbumin; Polylysine

In a previous study of the immunoregulatory properties of commensal bacterial DNA, we identified the strong immunostimulatory oligodeoxynucleotide (ISS-ODN) ID35 in the genomic DNA of Lactobacillus rhamnosus GG (LGG). The observed effects of ID35 are because of the unique TTTCGTTT motif located at the 5' end of the ODN, which is different from the previously identified ISS motifs in humans and mice. In the present study, we used an ovalbumin (OVA)-sensitized mouse model to show that ID35 is a potent suppressor of antigen-specific immunoglobulin E (IgE) production in vivo. This effect was toll-like receptor 9-dependent, as GpC negID35 failed to suppress antigen-specific IgE production. ID35 activated the specific subset of CD11c+CD8a+ dendritic cells, which are associated with T-helper 1 (Th1)-type systemic responses, and effectively induced interferon-gamma (IFN-gamma) production by CD4+ T cells in OVA-challenged mice. These immunoregulatory effects of ID35 were comparable with those induced by the murine prototype ODN 1826. Thus, ID35 is the first ISS-ODN with such a strong immunostimulatory and IgE suppressor activity to be found in immunobiotic bacterial DNA.

Topics: Allergens; Amino Acid Motifs; Animals; Antibody Specificity; CD11c Antigen; CD4-Positive T-Lymphocytes; CD8 Antigens; CHO Cells; Cricetinae; Cricetulus; Dendritic Cells; DNA, Bacterial; Hypersensitivity; Immunoglobulin E; Immunosuppression Therapy; Injections, Intraperitoneal; Interferon-gamma; Lacticaseibacillus rhamnosus; Mice; Mice, Inbred BALB C; Models, Animal; Oligodeoxyribonucleotides; Ovalbumin; Probiotics; Spleen; Toll-Like Receptor 9

Mycobacterium bovis Bacillus Calmette-Guérin (BCG) vaccination has been shown to inhibit allergic airway inflammation in animal models, associated with the regulation of allergen-specific T-cell immunity. However, little is known about whether neonatal BCG treatment could inhibit allergic inflammation by regulating allergen-specific T-cell response in aged mice. This study was aimed to investigate the impact of neonatal BCG treatment on allergic asthma and possible mechanism(s) underlying the action of BCG in different ages of mice.. C57BL/6 neonates were vaccinated with BCG on days 1, 7 and 14, sensitized with ovalbumin (OVA) at 5 and 7 weeks of age, and then challenged with allergen at 9 or 45 weeks of age for early- or late-challenged asthma. Their airway inflammation and allergen-specific T-cell responses were characterized.. Following early-challenge, BCG vaccination inhibited airway hyper-responsiveness (AHR), infiltration of eosinophils and mucous overproduction (P < 0.05), and shifted OVA-specific predominant Th2- to Th1-type cytokine responses in both the bronchoalveolar lavage fluid and the splenocyte supernatants (P < 0.05). In late-challenged mice, neonatal BCG treatment attenuated AHR and eosinophilia (P < 0.05), but failed to modulate allergen-specific cytokine responses.. Our data suggest that neonatal BCG vaccination has a long-term effect on inhibiting AHR and eosinophilia, which is associated with the modulation of Th1/Th2 cytokine production in early-, but not in late-challenged mice. Thus, different mechanisms may mediate the long-term protective effect of BCG neonatal vaccination differently in younger adult and aged mice.

Topics: Animals; Animals, Newborn; Asthma; BCG Vaccine; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Freeze Drying; Humans; Hypersensitivity; Inflammation; Mice; Mice, Inbred C57BL; Mycobacterium bovis; Ovalbumin; T-Lymphocytes

To experimentally examine the hygiene hypothesis, here we studied the effect of chlamydial infection on the development of allergic responses induced by OVA and the involvement of NK cells in this process using a mouse model of airway inflammation. We found that prior Chlamydia muridarum infection can inhibit airway eosinophilic inflammation and mucus production induced by allergen sensitization and challenge. The inhibition was correlated with an alteration of allergen-driven cytokine-producing patterns of T cells. We demonstrated that NK cells were activated following chlamydial infection, showing both cell expansion and cytokine secretion. The in vivo depletion of NK cells using anti-NK Ab before OVA sensitization and challenge partially abolished the inhibitory effect of chlamydial infection, which was associated with a partial restoration of Th2 cytokine production. In contrast, the adoptive transfer of NK cells that were isolated from infected mice showed a significant inhibitory effect on allergic responses, similar to that observed in natural infection. The data suggest that the innate immune cells such as NK cells may play an important role in infection-mediated inhibition of allergic responses.

Topics: Allergens; Animals; Cell Separation; Chlamydia muridarum; Cytokines; Female; Hypersensitivity; Killer Cells, Natural; Mice; Mice, Knockout; Mucus; Organic Anion Transport Protein 1; Ovalbumin; Pulmonary Eosinophilia; Th2 Cells; Vascular Cell Adhesion Molecule-1

Previous mouse and clinical studies demonstrate a link between Th2 intestinal inflammation and induction of the effector phase of food allergy. However, the mechanism by which sensitization and mast cell responses occurs is largely unknown. We demonstrate that interleukin (IL)-9 has an important role in this process. IL-9-deficient mice fail to develop experimental oral antigen-induced intestinal anaphylaxis, and intestinal IL-9 overexpression induces an intestinal anaphylaxis phenotype (intestinal mastocytosis, intestinal permeability, and intravascular leakage). In addition, intestinal IL-9 overexpression predisposes to oral antigen sensitization, which requires mast cells and increased intestinal permeability. These observations demonstrate a central role for IL-9 and mast cells in experimental intestinal permeability in oral antigen sensitization and suggest that IL-9-mediated mast cell responses have an important role in food allergy.

Topics: Administration, Oral; Anaphylaxis; Animals; Antigens; Cromolyn Sodium; Disease Susceptibility; Fatty Acid-Binding Proteins; Gene Expression Profiling; Hypersensitivity; Interleukin-9; Intestines; Mast Cells; Mastocytosis; Mice; Mice, Transgenic; Ovalbumin; Permeability; Phenotype; Rats; Receptors, Interleukin-4; STAT6 Transcription Factor; Th2 Cells

Allergic asthma is a chronic disease of the airways, with superimposed acute inflammatory episodes which correspond to exacerbations of asthma. Two novel models of allergic asthma have been developed in mice receiving the same allergen sensitisation, but with acute or chronic allergen exposures, the latter to mimic the human situation more closely. Ovalbumin-sensitised mice were challenged by ovalbumin inhalation twice on the same day for the acute model, and 18 times over a period of 6 weeks for the chronic model. Lung function was monitored in conscious, unrestrained mice immediately after the last challenge for up to 12 h. Airway responsiveness to inhaled methacholine and serum antibody levels were determined 24 h after challenge. Bronchoalveolar inflammatory cell recruitment was determined at 2 or 24 h. Acute and chronically treated mice had similar early and late asthmatic responses peaking at 2 h and 7-8 h, respectively. IgE and IgG antibody levels, compared with naïve mice, and eosinophil infiltration, compared with naïve and saline challenge, were elevated. Airway hyperresponsiveness to methacholine was observed 24 h after challenge in both models. The acute model had higher levels of eosinophilia, whereas the chronic model showed hyperresponsiveness to lower doses of methacholine and had higher levels of total IgE and ovalbumin-specific IgG antibodies. Both novel murine models of allergic asthma bear a close resemblance to human asthma, each offering particular advantages for studying the mechanisms underlying asthma and for evaluating existing and novel therapeutic agents.

Topics: Acute Disease; Administration, Inhalation; Allergens; Animals; Antibodies; Asthma; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Cell Count; Chronic Disease; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Phenotype; Plethysmography, Whole Body; Pneumonia; Respiratory Function Tests

We have shown previously that specific ligands of the peroxisome proliferator-activated receptor-gamma (PPARgamma) inhibit the systemic allergic immune response. The objective of this study was to investigate the impact of PPARgamma-ligand treatment on the local allergic immune response. We established a murine model exhibiting clinical and histological features of AD-like skin lesions with high reproducibility. In this model, the PPARgamma ligand was applied in an either preventive or therapeutic manner via systemic and local routes. The affected skin areas were assessed by standardized skin score, histological analyses, and immunohistochemical examinations. Our data show that systemic application of PPARgamma ligand by a preventive protocol led to significantly reduced onset of eczematous skin lesions. This was confirmed by histology, showing decreased skin thickness accompanied by significantly reduced infiltrations of CD4+ and CD8+ lymphocytes but also mast cells. Additionally, early allergen-specific IgE and IgG1 responses were reduced (day 21/35), whereas IgG2a levels remained unchanged. In conclusion, our results demonstrate that PPARgamma-ligand treatment inhibits not only systemic allergic immune response, but also local allergen-mediated dermatitis. Our findings point to therapeutic strategies, including a PPARgamma-ligand-based treatment.

Topics: Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Dermatitis, Atopic; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Ligands; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; Skin; Thiazolidinediones

We identify the tumor necrosis factor receptor superfamily 25 (TNFRSF25)/TNFSF15 pair as critical trigger for allergic lung inflammation, which is a cardinal feature of asthma. TNFRSF25 (TNFR25) signals are required to exert T helper cell 2 (Th2) effector function in Th2-polarized CD4 cells and co-stimulate interleukin (IL)-13 production by glycosphingolipid-activated NKT cells. In vivo, antibody blockade of TNFSF15 (TL1A), which is the ligand for TNFR25, inhibits lung inflammation and production of Th2 cytokines such as IL-13, even when administered days after airway antigen exposure. Similarly, blockade of TNFR25 by a dominant-negative (DN) transgene, DN TNFR25, confers resistance to lung inflammation in mice. Allergic lung inflammation-resistant, NKT-deficient mice become susceptible upon adoptive transfer of wild-type NKT cells, but not after transfer of DN TNFR25 transgenic NKT cells. The TNFR25/TL1A pair appears to provide an early signal for Th2 cytokine production in the lung, and therefore may be a drug target in attempts to attenuate lung inflammation in asthmatics.

Topics: Animals; Antibodies, Monoclonal; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cricetinae; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Lymphocyte Activation; Mice; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Receptors, Tumor Necrosis Factor, Member 25; Spleen; T-Lymphocytes; Th2 Cells

The effect of ovalbumin (Ova) sensitization on pulmonary C-fiber sensitivity was investigated. Brown-Norway rats were sensitized by intraperitoneal injection of Ova followed by aerosolized Ova three times per week for 3 wk. Control rats received the vehicle. At the end of the third week, single-unit fiber activities (FA) of pulmonary C fibers were recorded in anesthetized, artificially ventilated rats. Our results showed the following: 1) Ova sensitization induced airway inflammation (infiltration of eosinophils and neutrophils) and airway hyperresponsiveness in rats; 2) baseline FA in sensitized rats was significantly higher than that in control ones; 3) similarly, the pulmonary C-fiber response to right atrial injection of capsaicin was markedly higher in sensitized rats, which were significantly amplified after the acute Ova inhalation challenge; and 4) similar patterns, but smaller magnitudes of the differences in C-fiber responses to adenosine and lung inflation, were also found between sensitized and control rats. In conclusion, Ova sensitization elevated the baseline FA and excitability of pulmonary C fibers, and the hypersensitivity was further potentiated after the acute Ova inhalation challenge in sensitized rats. Chronic allergic inflammatory reactions in the airway probably contributed to the sensitizing effect on these lung afferents.

Topics: Administration, Inhalation; Aerosols; Allergens; Animals; Blood Pressure; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Dose-Response Relationship, Drug; Heart Rate; Hypersensitivity; Inflammation; Lung; Male; Methacholine Chloride; Nerve Fibers, Unmyelinated; Neurons, Afferent; Ovalbumin; Rats; Rats, Inbred BN

Epidemiological studies have shown an association between ambient particle inhalation and adverse respiratory heath effects. Inhalation of ultrafine particles (UFP, diameter <100 nm) has been suggested to contribute to exacerbation of allergic airway inflammation. Here we analyze the potential effects of allergen sensitization and challenge on total and regional deposition of UFP in the lung. Ovalbumin (OVA)-sensitized and nonsensitized mice were exposed for 1 h to ultrafine iridium particles radiolabeled with (192)Ir (UF-Ir) (0.2 mg m(-3)) at 2 different time points either before or after allergen (OVA) challenge. Additional sensitized and nonsensitized mice were exposed to UF-Ir without allergen challenge. Lung total and regional UF-Ir deposition were calculated according to the distribution of radioactivity in the body and in the excreta during 3 days following UF-Ir inhalation. OVA-sensitized mice showed a 21% relative increase of total UF-Ir deposited fraction compared to nonsensitized mice. When UF-Ir inhalation was performed after allergen challenge, no difference in total UF-Ir deposited fraction between sensitized and nonsensitized mice was detectable. Furthermore, no differences in extrathoracic deposition or in regional particle deposition were detected between all experimental groups. This study indicates that allergen sensitization alone can affect UFP deposition in the lungs. Whether higher UFP deposition in sensitized individuals compared to nonsensitized individuals or whether other factors, like alterations in long-term clearance kinetics, contribute substantially to the susceptibility of allergic individuals to particle exposure has yet to be elucidated.

Topics: Administration, Inhalation; Aerosols; Allergens; Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Hypersensitivity; Lung; Lung Diseases; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Particulate Matter; Random Allocation; Respiratory Function Tests

Farming has been widely reported to be associated with decreased risk of developing atopic disorders, but underlying immunomodulatory mechanisms are still not fully defined. We delineated T-cell functions after induction of mucosal tolerance in the context of intranasally delivered organic dust compounds, lipopolysaccharides (LPS).. BALB/c mice were pretreated intranasally with ovalbumin (OVA) with or without LPS (Escherichia coli) three times (days -21, -14, -7) prior to systemic OVA sensitization (days 1 and 14) and airway allergen challenges (days 28-30). CD4+ spleen T cells from pretreated and sensitized donors were characterized for cytokine function, and transferred into naive recipients prior to subsequent OVA sensitization and challenges.. Intranasal OVA pretreatment suppressed Th2-mediated immune and inflammatory responses and enhanced frequency of regulatory T cells in OVA-sensitized and -challenged mice. Addition of LPS to OVA, but not LPS alone, inhibited development of allergen-induced sensitization and eosinophilic airway infiltration, and markedly enhanced allergen-specific IgG1 serum levels and frequencies of IL-10- and IFN-gamma-producing CD4+ T cells. Transfer of CD4+ spleen T cells from OVA-pretreated animals protected naive recipients against subsequent allergen sensitization and airway disease, whereas transfer from LPS/OVA-pretreated animals only protected against allergen sensitization.. Microbial LPS modulated mucosal tolerance by inducing allergen-specific IgG1 production and distinct effector CD4+ T cells with a mixed regulatory/Th1 phenotype. Organic dust components such as LPS might therefore be important immune modulators in naturally occurring or preventive allergen-specific tolerance induction.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity; Immune Tolerance; Immunity, Mucosal; Immunoglobulin G; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Ovalbumin

T-bet and STAT4 play critical roles in helper T cell differentiation, especially for Th1 cells. However, it is still unknown about the relative importance and redundancy of T-bet and STAT4 for Th1 differentiation. It is also unknown about their independent role of T-bet and STAT4 in the regulation of allergic airway inflammation. In this study, we addressed these issues by comparing T-bet-deficient (T-bet(-/-)) mice, STAT4(-/-) mice, and T-bet- and STAT4-double-deficient (T-bet(-/-)STAT4(-/-)) mice on the same genetic background. Th1 differentiation was severely decreased in T-bet(-/-) mice and STAT4(-/-) mice as compared with that in wild-type mice, but Th1 differentiation was still observed in T-bet(-/-) mice and STAT4(-/-) mice. However, Th1 cells were hardly detected in T-bet(-/-)STAT4(-/-) mice. In contrast, the maintenance of Th17 cells was enhanced in T-bet(-/-) mice but was reduced in STAT4(-/-) mice and T-bet(-/-)STAT4(-/-) mice. In vivo, Ag-induced eosinophil and neutrophil recruitment into the airways was enhanced in T-bet(-/-) mice but was attenuated in STAT4(-/-) mice and T-bet(-/-)STAT4(-/-) mice. Ag-induced IL-17 production in the airways was also diminished in STAT4(-/-) mice and T-bet(-/-)STAT4(-/-) mice. These results indicate that STAT4 not only plays an indispensable role in T-bet-independent Th1 differentiation but also is involved in the maintenance of Th17 cells and the enhancement of allergic airway inflammation.

Topics: Animals; Bronchial Hyperreactivity; CD4-Positive T-Lymphocytes; Cell Differentiation; Cell Proliferation; Cytokines; Flow Cytometry; Hypersensitivity; Interleukin-17; Mice; Mice, Mutant Strains; Ovalbumin; Pneumonia; STAT4 Transcription Factor; T-Box Domain Proteins; T-Lymphocyte Subsets; Th1 Cells; Th2 Cells

Exposure to particulate matter (PM) air pollution is associated with increased asthma morbidity. Residual oil flash ash (ROFA) is rich in water-soluble transition metals, which are involved in the pathological effects of PM. The objective of this study was to investigate the effects of intranasal administration of ROFA on pulmonary inflammation, pulmonary responsiveness, and excess mucus production in a mouse model of chronic pulmonary allergic inflammation. BALB/c mice received intraperitoneal injections of ovalbumin (OVA) solution (days 1 and 14). OVA challenges were performed on days 22, 24, 26, and 28. After the challenge, mice were intranasally instilled with ROFA. After forty-eight hours, pulmonary responsiveness was performed. Mice were sacrificed, and lungs were removed for morphometric analysis. OVA-exposed mice presented eosinophilia in the bronchovascular space (p < .001), increased pulmonary responsiveness (p < .001), and epithelial remodeling (p = .003). ROFA instillation increased pulmonary responsiveness (p = .004) and decreased the area of ciliated cells in the airway epithelium (p = .006). The combined ROFA instillation and OVA exposure induced a further increase in values of pulmonary responsiveness (p = .043) and a decrease in the number of ciliated cells in the airway epithelium (p = .017). PM exposure results in pulmonary effects that are more intense in mice with chronic allergic pulmonary inflammation.

Topics: Air Pollutants; Animals; Carbon; Chronic Disease; Coal Ash; Disease Models, Animal; Hypersensitivity; Inhalation Exposure; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Particulate Matter; Pneumonia

Eosinophils have been implicated as playing a major role in allergic airway responses. However, the importance of these cells to the development of this disease has remained ambiguous despite many studies, partly because of lack of appropriate model systems. In this study, using transgenic murine models, we more clearly delineate a role for eosinophils in asthma. We report that, in contrast to results obtained on a BALB/c background, eosinophil-deficient C57BL/6 Delta dblGATA mice (eosinophil-null mice via the Delta DblGATA1 mutation) have reduced airway hyperresponsiveness, and cytokine production of interleukin (IL)-4, -5, and -13 in ovalbumin-induced allergic airway inflammation. This was caused by reduced T cell recruitment into the lung, as these mouse lungs had reduced expression of CCL7/MCP-3, CC11/eotaxin-1, and CCL24/eotaxin-2. Transferring eosinophils into these eosinophil-deficient mice and, more importantly, delivery of CCL11/eotaxin-1 into the lung during the development of this disease rescued lung T cell infiltration and airway inflammation when delivered together with allergen. These studies indicate that on the C57BL/6 background, eosinophils are integral to the development of airway allergic responses by modulating chemokine and/or cytokine production in the lung, leading to T cell recruitment.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Eosinophils; GATA1 Transcription Factor; Hypersensitivity; Inflammation; Interleukins; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Species Specificity; T-Lymphocytes

Successful allergen-specific immunotherapy (SIT) is associated with reduced Th2 cytokine production and the induction of IL-10-producing regulatory T cells. To improve treatment efficacy, we investigated the impact of an IL-4/IL-13 inhibitor during SIT.. BALB/c mice were sensitized intranasally with ovalbumin (OVA) for 4 weeks. Subsequently, they were subjected to intranasal SIT, with OVA being administered at doses increasing from 1 mug to 1 mg over 3 weeks with or without an IL-4/IL-13 inhibitor. Serum OVA-specific antibodies were measured and bronchoalveolar lavage (BAL) fluids were checked for airway eosinophilia. Subsequently, lung tissue was examined histologically for inflammatory infiltrates. Cytokines were detected in BAL fluids and spleen cell cultures. Furthermore, CD4 CD25 double-positive spleen T cells were checked for intracellular IL-10 production by flow cytometry.. OVA sensitization resulted in persistent IgE synthesis and an eosinophil-rich allergic airway inflammation combined with increased IL-4 and IL-5 levels. Therefore, intranasal SIT could efficiently reverse the allergic phenotype. This was associated with decreased IL-4 and IL-5 levels, and increased IL-10 levels in BAL fluids as well as increased amounts of IL-10-producing CD25+ regulatory T cells. However, mice treated with the IL-4/IL-13 inhibitor during SIT did not produce significantly different results .. The use of an IL-4/IL-13 inhibitor as an adjuvant for SIT did not enhance anti-allergic effects. Thus, the observed reversal of Th2 responses during SIT may not be the keystone for successful therapy, but rather other factors, e.g. IL-10-producing regulatory T cells, may be crucial.

Topics: Administration, Intranasal; Animals; Asthma; Bronchoalveolar Lavage Fluid; Desensitization, Immunologic; Enzyme-Linked Immunosorbent Assay; Eosinophils; Flow Cytometry; Hypersensitivity; Inflammation; Interleukin-10; Interleukin-13; Interleukin-2 Receptor alpha Subunit; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

The edeines analogs were tested in several in vitro and in vivo assays using the mouse model, with edeine B (peptide W1) and cyclosporine A as reference compounds. The peptides displayed moderate, stimulatory effects on concanavalin A-induced (ConA-induced) splenocyte proliferation, whereas their effects on pokeweed mitogen-induced (PWM-induced) splenocyte proliferation were inhibitory. The peptides inhibited lipopolysacharide-induced (LPS-induced) tumor necrosis factor alpha production but had little effect on interleukin 6 production. In the model of the humoral immune response in vitro to sheep red blood cells, peptide 1 was distinctly stimulatory in the investigated concentrations (1-100 microg/ml), whereas peptides 3 and 4 only stimulated the number of antibody-forming cells at the highest concentration (100 microg/ml). In the model of the delayed type hypersensitivity in vivo to ovalbumin, the peptides were moderately suppressive (3 being the most active). The reference peptide W1 stimulated ConA-induced cell proliferation at 1-10 microg/ml but was inhibitory at 100 microg/ml. It also inhibited PWM-induced cell proliferation in a dose-dependent manner. This peptide had no effect on the humoral immune response in vitro or on cytokine production, but inhibited DTH reaction in vivo. The relationship between structure and activity, and a possible mode of action of the peptides, is discussed in this paper.

Topics: Animals; Cell Proliferation; Concanavalin A; Edeine; Hypersensitivity; Immunity; Interleukin-6; Lipopolysaccharides; Mice; Ovalbumin; Pokeweed Mitogens; Sheep; Spleen; Tumor Necrosis Factor-alpha

Although many studies regarding airway remodeling in asthma have been reported, only a few studies have investigated airway remodeling in allergic rhinitis.. To determine whether repetitive allergen challenge could induce airway remodeling in the nose and evaluate the effect of steroids using a murine model of allergic rhinitis.. To develop a mouse model of airway remodeling, ovalbumin-sensitized mice were repeatedly exposed to inhaled ovalbumin administration twice a week for 1 month and 3 months. Matched control mice were challenged with phosphate-buffered saline, and the treatment group received intraperitoneal dexamethasone injection. Trichrome, periodic acid-Schiff, hematoxylin-eosin, and immunohistochemical staining against matrix metalloproteinase 9 and tissue inhibitors of metalloproteinase 1 were performed to nasal and lung tissues, and the level of transforming growth factor beta in the nasal lavage fluid was analyzed.. Repetitive ovalbumin challenge for 3 months induced circumferential peribronchial fibrosis in the lung. In the nose, subepithelial fibrosis, increased matrix metalloproteinase 9 and tissue inhibitors of metalloproteinase 1 expression, goblet cell hyperplasia, and submucous gland hypertrophy were observed compared with the control group. Features of airway remodeling were more prominent in the lung tissue. Administration of dexamethasone significantly inhibited these histologic changes.. Airway remodeling associated with long-term allergen challenge can occur in the nasal mucosa and the lung. Steroid treatment prevents airway inflammation in response to acute allergen challenge, as well as airway remodeling by long-term allergen challenge.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Dexamethasone; Fibrosis; Hypersensitivity; Immunohistochemistry; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Rhinitis; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta

Several studies indicate that selectin-mediated leukocyte migration may depend on the types of initiating inflammatory stimuli or on the vascular beds involved in the inflammatory response. Thus, targeting selectin interactions to treat inflammation may have variable effects depending on the site and origin of the inflammatory response.. To address whether selectin-mediated leukocyte recruitment is stimulus or tissue dependent.. We examined pulmonary and cutaneous allergic inflammatory responses and silica-induced nonallergic lung inflammation and fibrosis in wild-type and P- and E-selectin-deficient (P/E-/-) double knockout mice. Allergen-sensitized wild-type and P/E-/- double knockout mice were challenged either intradermally or via the airways to induce allergic responses in the skin or lung, respectively. Other animals were subjected to intranasal silica administration to induce a nonallergic lung inflammatory/fibrotic response.. The P/E-/- mice exhibited significantly reduced allergic inflammation in the skin and lung. Allergic late-phase ear swelling and allergic lung airway hyperresponsiveness were also significantly attenuated in the P/E-/- mice compared with identically treated wild-type animals. In contrast, pulmonary inflammation and fibrosis induced by intranasal administration of silica particles resulted in a more severe phenotype in the P/E-/- mice.. Selectin interactions drive allergic inflammation in the lung and skin. Silica-induced pulmonary inflammation and fibrosis, however, was more pronounced in the absence of selectin interactions, suggesting that selectin-mediated leukocyte migration may depend on the types of initiating inflammatory stimuli.

Topics: Animals; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Cytokines; E-Selectin; Endothelium, Vascular; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; P-Selectin; Silicon Dioxide; Skin

Since liposomes are known as strong adjuvants, we attempted to use liposomes in immunotherapy as adjuvants, and to achieve desensitization in pre-sensitized mice. At first, we sensitized mice with intraperitoneal injection of model antigen, 100 microg ovalbumin (OVA), with Alum and treated them with liposome composed of distearoylphosphatidylcholine (DSPC) and cholesterol (2:1 as a molar ratio), which was coupled with a small amount of OVA (10 microg OVA in 400 nmol DSPC and 200 nmol cholesterol-liposome was injected into 20 g mouse). It is well known that antigen-specific immunotherapy increases IgG blocking antibodies and decreases in IgE antibodies. The treatment with i.v. injection of OVA-liposome at days 8, 10, and 12 after sensitization strongly suppressed OVA-specific IgE production without affecting IgG level after the boost (100 microg OVA with Alum). Moreover, the treatment with high-density OVA-liposome (10 microg OVA in 80 nmol DSPC and 40 nmol cholesterol-liposome/20 g mouse) not only strongly suppressed IgE levels but also reduced IgG production after the boost of OVA-sensitized mice suggesting the importance of liposomal characteristic in desensitization immunotherapy. Next we reduced the dose of OVA-liposome and the desensitization effect was also observed at the dose of as low as 1 microg OVA on OVA-liposome/mouse. On the contrary, free OVA did not affect the production of both IgG and IgE levels. Biodistribution study indicated that OVA-liposome was highly accumulated in spleen of OVA-sensitized mice compared to control liposome at 3 h after i.v. injection. These results suggest that the liposomal OVA effectively interacts with and desensitizes immune cells, therefore, liposomes coupling with a certain antigen may be effective in allergy immunotherapy.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Antigens; Cholesterol; Desensitization, Immunologic; Dose-Response Relationship, Immunologic; Drug Administration Schedule; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Liposomes; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphatidylcholines; Spleen; Tissue Distribution

Experimental airway allergy in mice leads to increased activity in specific hypothalamic and amygdaloid nuclei, and behavioral changes. The experiments described here were designed to determine the role of anaphylactic antibodies, mast cell degranulation, and lung inflammation in the neural and behavioral correlates of an experimental murine asthma-like response. Animals were sensitized intraperitoneally with ovalbumin adsorbed to alum, and challenged by intranasal ovalbumin instillation or aerosol. To induce immunological tolerance, animals were fed ovalbumin in the drinking water for 5 consecutive days, along with primary sensitization. Depletion of IgE was also accomplished with a non-anaphylactic anti-IgE antibody. Mast cell degranulation was inhibited by cromolyn. In addition to BALB/c animals, C3H/HeJ mice were used for their relative resistance to lung allergic inflammation. We confirmed that ovalbumin challenge in allergic mice leads to increased activity in the paraventricular nucleus of the hypothalamus and central nucleus of the amygdala, and avoidance behavior towards an allergen-associated compartment. Moreover, these responses were precluded by oral tolerance or anti-IgE treatment, even in the presence of IgG1. Cromolyn abrogates both responses in the presence of anaphylactic antibodies. Finally, although sensitized C3H/HeJ mice did not develop airway inflammation, they exhibited brain and behavioral changes similar to BALB/c animals. The repercussions of murine allergic asthma on brain and behavior are IgE-dependent, mediated by mast cell degranulation, and do not require a pulmonary inflammatory infiltrate, suggesting that the early phase of this immediate allergic response suffices for the brain activation associated with avoidance behavior towards exposure to the allergen.

Topics: Amygdala; Analysis of Variance; Anaphylaxis; Animals; Asthma; Avoidance Learning; Behavior, Animal; Cell Degranulation; Disease Models, Animal; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Male; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Neuroimmunomodulation; Ovalbumin; Paraventricular Hypothalamic Nucleus; Species Specificity; Statistics, Nonparametric

Epidemiological data have indicated that some infections are associated with a low risk of allergic diseases, thus supporting the idea (hygiene hypothesis) that the microbial load is an important environmental factor conferring protection against the development of allergies. We set out to test the hygiene hypothesis in a unique epidemiological setting in two socio-economically and culturally markedly different, although genetically related, populations living in geographically adjacent areas. The study cohorts included 266 schoolchildren from the Karelian Republic in Russia and 266 schoolchildren from Finland. The levels of total IgE and allergen-specific IgE for birch, cat and egg albumen were measured. Microbial antibodies were analysed against enteroviruses (coxsackievirus B4), hepatitis A virus, Helicobacter pylori and Toxoplasma gondii. Although total IgE level was higher in Russian Karelian children compared to their Finnish peers, the prevalence of allergen-specific IgE was lower among Russian Karelian children. The prevalence of microbial antibodies was, in turn, significantly more frequent in the Karelian children, reflecting the conspicuous difference in socio-economic background factors. Microbial infections were associated with lower risk of allergic sensitization in Russian Karelian children, enterovirus showing the strongest protective effect in a multivariate model. The present findings support the idea that exposure to certain infections, particularly in childhood, may protect from the development of atopy. Enterovirus infections represent a new candidate to the list of markers of such a protective environment. However, possible causal relationship needs to be confirmed in further studies.

Topics: Adolescent; Allergens; Animals; Antibodies, Bacterial; Antibodies, Protozoan; Antibodies, Viral; Bacteria; Betula; Cats; Child; Enterovirus B, Human; Female; Finland; Helicobacter pylori; Hepatitis A virus; Humans; Hypersensitivity; Immunoglobulin E; Male; Ovalbumin; Pollen; Russia; Toxoplasma; Viruses

There are no studies examining the effects of sevoflurane on a chronically inflamed and remodeled airway, such as that found in asthma. In the present study, we sought to define the respiratory effects of sevoflurane in a model of chronic allergic asthma. For this purpose, pulmonary mechanics were studied and lung morphometry analyzed to determine whether the physiological modifications reflected underlying morphological changes.. Thirty-six BALB/c mice (20-25 g) were randomly divided into four groups. In OVA groups, mice were sensitized with ovalbumin and exposed to repeated ovalbumin challenges. In SAL groups, mice received saline using the same protocol. Twenty-four hours after the last challenge, the animals were anesthetized with pentobarbital sodium (PENTO, 20 mg/kg i.p.) or sevoflurane (SEVO, 1 MAC). Lung static elastance (Est), resistive ([DELTA]P1) and viscoelastic/inhomogeneous ([DELTA]P2) pressure decreases were analyzed by an end-inflation occlusion method. Lungs were fixed and stained for histological analysis.. Animals in the OVASEVO group showed lower [DELTA]P1 (38%), [DELTA]P2 (24%), and Est (22%) than animals in the OVAPENTO group. Histology demonstrated greater airway dilation (16%) and a lower degree of alveolar collapse (25%) in the OVASEVO compared with OVAPENTO group. [DELTA]P1 was lower (35%) and airway diameters larger (12%) in the SALSEVO compared with SALPENTO group.. Sevoflurane anesthesia acted both at airway level and lung periphery reducing ([DELTA]P1 and [DELTA]P2 pressures, and Est in chronic allergic asthma.

Topics: Anesthetics, Inhalation; Animals; Asthma; Disease Models, Animal; Hypersensitivity; Lung; Methyl Ethers; Mice; Mice, Inbred BALB C; Ovalbumin; Pressure; Respiration; Respiratory Mechanics; Sevoflurane; Time Factors

Adrenomedullin (ADM), a newly identified vasodilating peptide, is reported to be expressed in lungs and have a bronchodilating effect. We hypothesized whether ADM could be involved in the pathogenesis of bronchial asthma. We examined the role of ADM in airway responsiveness using heterozygous ADM-deficient mice (AM+/-) and their littermate control (AM+/+). Here, we show that airway responsiveness is enhanced in ADM mutant mice after sensitization and challenge with ovalbumin (OVA). The immunoreactive ADM level in the lung tissue after methacholine challenge was significantly greater in the wild-type mice than that in the mutant. However, the impairment of ADM gene function did not affect immunoglobulins (OVA-specific IgE and IgG1), T helper 1 and 2 cytokines, and leukotrenes. Thus the conventional mechanism of allergen-induced airway responsiveness is not relevant to this model. Furthermore, morphometric analysis revealed that eosinophilia and airway hypersecretion were similarly found in both the OVA-treated ADM mutant mice and the OVA-treated wild-type mice. On the other hand, the area of the airway smooth muscle layer of the OVA-treated mutant mice was significantly greater than that of the OVA-treated wild-type mice. These results suggest that ADM gene disruption may be associated with airway smooth muscle hyperplasia as well as enhanced airway hyperresponsiveness. ADM mutant mice might provide novel insights to study the pathophysiological role of ADM in vivo.

Topics: Adrenomedullin; Allergens; Animals; Asthma; Bronchial Provocation Tests; Hypersensitivity; Mice; Mice, Knockout; Ovalbumin

These studies were conducted to investigate the role of dermal exposure to perfluorooctanoic acid (PFOA), a known immunosuppressant, on the hypersensitivity response to ovalbumin (OVA) in a murine model of asthma. PFOA has had widespread use as a carpet and fabric protectant. BALB/c mice were exposed dermally, on the dorsal surface of each ear, to concentrations of PFOA ranging from 0.01 to 1.5% (applied dose 0.25-50 mg/kg) for 4 days. In hypersensitivity studies, mice were also ip injected with 7.5 microg OVA and 2 mg alum on days 1 and 10 and in some studies challenged with 250 microg OVA by pharyngeal aspiration on days 17 and 26. Following exposure to PFOA, an increase in liver weights and a decrease in thymus and spleen weights and cellularities were observed. Similar immunomodulatory trends were demonstrated in mice coadministered PFOA and OVA. Compared to the OVA alone-exposed animals, an increase in total IgE was demonstrated when mice were coexposed to OVA and concentrations of PFOA ranging from 0.75 to 1.5%, while the OVA-specific IgE response peaked with 0.75% PFOA coexposure (p < or = 0.05). OVA-specific airway hyperreactivity was increased in the 1.0% PFOA coexposed group (p < or = 0.05), with an increased pleiotropic cell response characterized by eosinophilia and mucin production, in animals coexposed to concentrations of PFOA up to 1.0%, as compared to the OVA alone-exposed animals. In a murine model, PFOA was demonstrated to be immunotoxic following dermal exposure, with an enhancement of the hypersensitivity response to OVA, suggesting that PFOA exposure may augment the IgE response to environmental allergens.

Topics: Administration, Topical; Animals; Antigens; Asthma; Body Weight; Bronchial Hyperreactivity; Caprylates; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Fluorocarbons; Hypersensitivity; Immunoglobulin E; Immunosuppressive Agents; Lung; Mice; Mice, Inbred BALB C; Organ Size; Ovalbumin; Phenotype; Respiratory Hypersensitivity

Chronic allergic asthma is associated with marked inflammatory reaction, microvascular leakage and epithelium injury. As previously shown in a rat model of chronic asthma, these alterations increase lung permeability and distal airway fluid clearance. Keratinocyte growth factor (KGF) has been shown to induce epithelial cell proliferation and to protect from acute lung injuries. Therefore, the current authors evaluated the potential role of KGF treatment on lung permeability and airway inflammation in rats with chronic asthma. KGF (1 mg x kg(-1)) was administered intravenously before the last ovalbumin (OVA) challenge in sensitised rats. Permeability was assessed by the leak of radiolabelled albumin from the alveolar and systemic compartments. Histopathological analysis was also performed. Treatment with KGF decreased the leak of both markers and decreased the level of extravascular lung water in sensitised rats challenged with OVA. KGF treatment also reduced the inflammatory cell number in bronchoalveolar lavage fluid but not in bronchial mucosa. KGF markedly limited the allergen-induced alterations in epithelium integrity and the expression of the intercellular junction proteins beta-catenin and zonula occludens protein-1. In conclusion, keratinocyte growth factor administration markedly limits lung permeability and airway inflammation, an effect associated with a decrease in epithelium alterations during chronic allergic asthma. These data open new prospects in the therapeutic strategy of asthma.

Topics: Animals; Asthma; beta Catenin; Bronchi; Epithelial Cells; Epithelium; Fibroblast Growth Factor 7; Hypersensitivity; Inflammation; Lung; Male; Mucous Membrane; Ovalbumin; Permeability; Rats

Osteopontin (Opn) is important for T helper type 1 (T(H)1) immunity and autoimmunity. However, the role of this cytokine in T(H)2-mediated allergic disease as well as its effects on primary versus secondary antigenic encounters remain unclear. Here we demonstrate that OPN is expressed in the lungs of asthmatic individuals and that Opn-s, the secreted form of Opn, exerts opposing effects on mouse T(H)2 effector responses and subsequent allergic airway disease: pro-inflammatory at primary systemic sensitization, and anti-inflammatory during secondary pulmonary antigenic challenge. These effects of Opn-s are mainly mediated by the regulation of T(H)2-suppressing plasmacytoid dendritic cells (DCs) during primary sensitization and T(H)2-promoting conventional DCs during secondary antigenic challenge. Therapeutic administration of recombinant Opn during pulmonary secondary antigenic challenge decreased established T(H)2 responses and protected mice from allergic disease. These effects on T(H)2 allergic responses suggest that Opn-s is an important therapeutic target and provide new insight into its role in immunity.

Topics: Animals; Anti-Inflammatory Agents; Bronchi; Dendritic Cells; Disease Models, Animal; Humans; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Osteopontin; Ovalbumin; Recombinant Proteins

The contributing role of indoor air pollution to the development of allergic disease has become increasingly evident in public health problems. It has been reported that extensive communication exists between neurons and immune cells, and neurotrophins are molecules potentially responsible for regulating and controlling this neuroimmune crosstalk. The adverse effects of volatile organic compounds which are main indoor pollutants on induction or augmentation of neuroimmune interaction have not been fully characterized yet. To investigate the effects of low-level toluene inhalation on the airway inflammatory responses, male C3H mice were exposed to filtered air (control), 9 ppm, and 90 ppm toluene for 30 min by nose-only inhalation on Days 0, 1, 2, 7, 14, 21, and 28. Some groups of mice were injected with ovalbumin intraperitoneally before starting exposure schedule and these mice were then challenged with aerosolized ovalbumin as booster dose. For analysis of airway inflammation, bronchoalveolar lavage (BAL) fluid were collected to determine inflammatory cell influx and lung tissue and blood samples were collected to determine cytokine and neurotrophin mRNA and protein expressions and plasma antibody titers using real-time RT-PCR and ELISA methods respectively. Exposure of the ovalbumin-immunized mice to low-level toluene resulted in (1) increased inflammatory cells infiltration in BAL fluid; (2) increased IL-5 mRNA, decreased nerve growth factor receptor tropomyosin-related kinase A and brain-derived neurotrophic factor mRNAs in lung; and (3) increased IgE and IgG(1) antibodies and nerve growth factor content in the plasma. These findings suggest that low-level toluene exposure aggravates the airway inflammatory responses in ovalbumin-immunized mice by modulating neuroimmune crosstalk.

Topics: Administration, Inhalation; Animals; Bronchoalveolar Lavage Fluid; Corticosterone; Cytokines; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lung; Male; Mice; Mice, Inbred C3H; Nerve Growth Factors; Neurotoxicity Syndromes; Ovalbumin; Receptor Cross-Talk; Receptors, Nerve Growth Factor; Respiratory Hypersensitivity; RNA, Messenger; Solvents; Toluene

There have been a number of reports showing that the crude beta-glucan fraction prepared from various kinds of Basidiomycetes mushrooms acts as anti-cancer and anti-allergic reagent through stimulation of IFN-gamma production. It has been reported, however, that the exposure of the airway to beta-(1,3) d-glucan, contained in house dust, indoor moulds and some bacteria, potentiates the airway allergic response. It seems likely that the discrepant effects on immune function may be related to such factors as differences of the administration route, average molecular weight and water solubility. We isolated a new low-molecular-weight (about 100 kDa) beta-glucan from Aureobasidium pullulans 1A1 strain of black yeast, and found that it had low viscosity and was water-soluble. In this study, we examined the effects of water-soluble low-molecular-weight beta-(1-->3) and 50-80% branched beta-(1-->6) glucan (LMW-beta-Glucan) isolated from A. pullulans on the ova-albumin (OVA)-treated allergic reaction in mice. Feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan significantly inhibited the OVA-specific IgE elevation compared to that in OVA-sensitized mice fed standard laboratory diet alone (control). Furthermore, feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan inhibited the reduction of IL-12 and IFN-gamma production from splenocytes and the reduction of CD8- and IFN-gamma-positive cell number in the small intestine of the OVA-sensitized mice. These findings suggest that anti-food allergic action of LMW-beta-Glucan may be due to the inducing IFN-gamma production in the small intestine and splenocytes.

Topics: Animal Feed; Animals; Anti-Allergic Agents; Antineoplastic Agents; Basidiomycota; Carbohydrate Sequence; Cytokines; Dose-Response Relationship, Drug; Down-Regulation; Food Hypersensitivity; Glucans; Hypersensitivity; Immunoglobulin E; Immunologic Factors; Male; Mice; Mice, Inbred BALB C; Molecular Weight; Ovalbumin; Solubility; Spleen; Water

The objective of this study was to assess the antiallergic effect of fermented milk prepared, respectively, with Streptococcus thermophilus MC, Lactobacillus acidophilus B, Lactobacillus bulgaricus Lb, L. bulgaricus 448, and Bifidobacterium longum B6. Female BALB/c mice fed fermented milk were immunized intraperitoneally with ovalbumin (OVA)/complete Freund's adjuvant (CFA) to evaluate the immune response by observing the secretion of cytokines IL-2, IL-4, and IFN-gamma and serum antibody IgE. The results showed that supplementation with lactic acid bacteria fermented milk did not significantly change the IL-2 spontaneous and OVA-stimulated secretions of splenocytes. However, both spontaneous and OVA-stimulated secretions of splenocytes from mice fed lactic acid bacteria fermented milk showed significantly (P < 0.05) lower levels of IL-4 (Th2 cytokine) than those from OVA/CFA-immunized mice fed non-fermented milk (OVA/CFA-milk group). The spontaneous secretion of IFN-gamma (Th1 cytokine) by splenocytes from mice fed L. bulgaricus 448 or L. bulgaricus Lb fermented milk significantly increased as compared to that from the OVA/CFA-milk group. The results showed that the ratios of IFN-gamma to IL-4 of both spontaneous and OVA-stimulated secretions in splenocytes from mice fed lactic acid bacteria fermented milk increased significantly as compared to that of PBS- or OVA/CFA-milk groups. The serum levels of OVA-specific IgE in fermented milk fed groups, especially the group fed S. thermophilus MC fermented milk, were significantly lower than those in the OVA/CFA-milk group through a 6 week feeding experiment. The results showed that milk fermented with lactic acid bacteria demonstrated in vivo antiallergic effects on OVA/CFA-immunized mice via increasing the secretion ratio of IFN-gamma/IL-4 (Th1/Th2) by splenocytes and decreasing the serum level of OVA-specific IgE.

Topics: Animals; Female; Fermentation; Hypersensitivity; Immunoglobulin E; Lactobacillus acidophilus; Mice; Mice, Inbred BALB C; Milk; Models, Animal; Ovalbumin; Probiotics

Chronic inflammation in asthma and chronic obstructive pulmonary disease drives pathological structural remodelling of the airways. Using tiotropium bromide, acetylcholine was recently identified as playing a major regulatory role in airway smooth muscle remodelling in a guinea pig model of ongoing allergic asthma. The aim of the present study was to investigate other aspects of airway remodelling and to compare the effectiveness of tiotropium to the glucocorticosteroid budesonide. Ovalbumin-sensitised guinea pigs were challenged for 12 weeks with aerosolised ovalbumin. The ovalbumin induced airway smooth muscle thickening, hypercontractility of tracheal smooth muscle, increased pulmonary contractile protein (smooth-muscle myosin) abundance, mucous gland hypertrophy, an increase in mucin 5 subtypes A and C (MUC5AC)-positive goblet cell numbers and eosinophilia. It was reported previously that treatment with tiotropium inhibits airway smooth muscle thickening and contractile protein expression, and prevents tracheal hypercontractility. This study demonstrates that tiotropium also fully prevented allergen-induced mucous gland hypertrophy, and partially reduced the increase in MUC5AC-positive goblet cell numbers and eosinophil infiltration. Treatment with budesonide also prevented airway smooth muscle thickening, contractile protein expression, tracheal hypercontractility and mucous gland hypertrophy, and partially reduced MUC5AC-positive goblet cell numbers and eosinophilia. This study demonstrates that tiotropium and budesonide are similarly effective in inhibiting several aspects of airway remodelling, providing further evidence that the beneficial effects of tiotropium bromide might exceed those of bronchodilation.

Topics: Adrenal Cortex Hormones; Allergens; Animals; Bronchodilator Agents; Budesonide; Cholinergic Antagonists; Eosinophilia; Extracellular Matrix; Glucocorticoids; Guinea Pigs; Humans; Hypersensitivity; Inflammation; Male; Muscle, Smooth; Ovalbumin; Scopolamine Derivatives; Tiotropium Bromide; Trachea

Relaxin is a reproductive hormone with pleiotropic actions. In addition to airway fibrosis, relaxin deficiency results in airway structural changes (epithelial thickening) and increased lung recoil, suggesting that relaxin may impact other aspects of airway/lung structure and function beyond its ability to regulate collagen turnover. Furthermore, these structural changes associated with relaxin deficiency show marked similarity to the structural changes seen in asthma. The current study investigated the broader role of relaxin in regulating airway structure and function and examined the relationship between airway inflammation, structural changes, and airway hyperresponsiveness (AHR) using an ovalbumin (OVA)-induced model of allergic airways disease (AAD). The model of AAD was applied to 12-month-old relaxin-deficient (Rln(-/-)) mice with established airway fibrosis and age-matched wild-type (Rln(+/+)) controls. OVA-treated Rln(+/+) mice (induced inflammation) developed increased epithelial thickening (P < 0.05) and AHR (P < 0.05) but not airway fibrosis, compared with saline-treated Rln(+/+) controls. Saline-treated Rln(-/-) mice had significantly increased lung collagen deposition (existing fibrosis) and epithelial thickening and remarkably were found to have increased AHR that was equivalent to that in OVA-treated Rln(+/+) mice (all P < 0.05 vs. saline-treated Rln(+/+) controls). OVA-treated Rln(-/-) mice (existing fibrosis and induced inflammation) had increased airway/lung fibrosis (P < 0.05) but equivalent airway inflammation and AHR compared with OVA-treated Rln(+/+) animals. These findings demonstrate for the first time a role for relaxin in the regulation of airway responses using Rln(-/-) mice and suggest that airway fibrosis and/or epithelial thickening can result in increased AHR equivalent to that induced by airway inflammation in AAD.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Relaxin; Respiratory Physiological Phenomena; Respiratory System

Allergen exclusion measures during pregnancy and lactation have been given consideration in studies of primary allergy prevention but complete avoidance of mother/neonatal allergen exposure has proven to be a difficult procedure. To evaluate a strategy to prevent allergen sensitization in early life in mice, we first established a neonatal model with ovalbumin sensitization through maternal allergen exposure during pregnancy or breastfeeding. The modulatory potential of preconception immunization was investigated on the neonatal development of subsequent allergic responses to maternal allergen exposure. Herein, we demonstrate that immunized mothers exposed to antigen during pregnancy or breastfeeding underwent intense vertical transmission of antibodies, including immunoglobulin G (IgG) in complex with ovalbumin and IgG1 antibody with anaphylactic function. It was further shown that maternal immunization efficiently decreased the passage of free antigens through breastfeeding and inhibited the enhanced IgE antibody response after postnatal antigen exposure. In addition, antenatal immunization decreased the antigen-specific proliferative response of immunized neonates, in parallel with profound downmodulatory effects on both the activation and differentiation of T and B cells after a non-specific stimulus and cytokine production. These findings showed that early life sensitization, subsequent to maternal allergen exposure during both the prenatal and postnatal periods, could be avoided by preventive vaccination of the mother.

Topics: Allergens; Animals; Animals, Newborn; Cytokines; Female; Hypersensitivity; Immunity, Maternally-Acquired; Immunization; Immunoglobulin E; Lactation; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Milk; Ovalbumin; Preconception Care; Pregnancy

Parental phenotype is known to influence the inheritance of atopic diseases, such as allergic asthma, with a maternal history being a more significant risk factor for progeny than paternal history. We hypothesized that recall Th1- or Th2-type immune responses during pregnancy would result in transfer of maternal factors that would differentially impact development of immune responsiveness in offspring. Following weaning, susceptibility and severity of allergic airway disease (a murine model of human asthma) was evaluated in progeny, disease being elicited by immunization with OVA-Al(OH)(3) and challenge with aerosolized OVA. We found that progeny of mothers with Th1-biased immunity to OVA subjected to recall aerosol challenge during pregnancy had reduced levels of Ag-specific IgE and airway eosinophilia compared with progeny of mothers with Th2-biased immunity to OVA or naive mothers. Interestingly, progeny of mothers with Th1-type immunity to a heterologous albumin, BSA, were not protected from developing OVA-induced allergic airway disease. These findings demonstrated that maternal transfer of protection from development of allergic airway disease to offspring in this model of maternal Th1-type immunity was Ag specific.

Topics: Adjuvants, Immunologic; Animals; Asthma; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity; Immune System; Immunoglobulins; Male; Mice; Ovalbumin; Pregnancy; Th1 Cells; Th2 Cells

We evaluated the role of estradiol and progesterone in allergic lung inflammation. Rats were ovariectomized (Ovx) and, 7 days later, were sensitized with ovalbumin (OA) and challenged after 2 wk with inhaled OA; experiments were performed 1 day thereafter. Ovx-allergic rats showed reduced cell recruitment into the bronchoalveolar lavage (BAL) fluid relative to sham-Ovx allergic rats, as was observed in intact allergic rats treated with ICI-182,780. Estradiol increased the number of cells in the BAL of Ovx-allergic rats, whereas progesterone induced an additional reduction. Cells of BAL and bone marrow (BM) of Ovx-allergic rats released elevated amounts of IL-10 and reduced IL-1beta and TNF-alpha. BM cells of Ovx-allergic rats released increased amounts of IL-10 and lower amounts of IL-4. Estradiol treatment of Ovx-allergic rats decreased the release of IL-10 but increased that of IL-4 by BM cells. Estradiol also caused an increased release of IL-1beta and TNF-alpha by BAL cells. Progesterone significantly increased the release of IL-10, IL-1beta, and TNF-alpha by BAL cells and augmented that of IL-4 by BM cells. Degranulation of bronchial mast cells from Ovx rats was reduced after in vitro challenge, an effect reverted by estradiol but not by progesterone. We suggest that the serum estradiol-to-progesterone ratio might drive cellular recruitment, modulating the pulmonary allergy and profile of release of anti-inflammatory or inflammatory cytokines. The existence of such dual hormonal effects suggests that the hormone therapy of asthmatic postmenopausal women and of those suffering of premenstrual asthma should take into account the possibility of worsening the pulmonary conditions.

Topics: Animals; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Cell Degranulation; Cytokines; Disease Models, Animal; Estradiol; Female; Hypersensitivity; Interleukin-10; Interleukin-1beta; Interleukin-4; Leukocyte Count; Mast Cells; Ovalbumin; Ovariectomy; Pneumonia; Progesterone; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

Creatine supplement is the most popular nutritional supplement, and has various metabolic functions and sports medicine applications. Creatine supplementation increases muscle mass and can decrease muscular inflammation. Some studies have also suggested a beneficial role of creatine supplementation on chronic pulmonary diseases such as chronic obstructive pulmonary disease and cystic fibrosis. Among athletes, the prevalence of asthma is high, and many of these individuals may be taking creatine. However, the effects of creatine supplementation on chronic pulmonary diseases of allergic origin have not been investigated. In the present study, we analyzed the effects of creatine supplementation on a model of chronic allergic lung inflammation. Thirty-one Balb/c mice were divided into four groups: control, creatine (Cr), ovalbumin (OVA), and OVA+Cr. OVA and OVA+Cr groups were sensitized with intraperitoneal injections of OVA on Days 0, 14, 28, and 42. OVA challenge (OVA 1%) and Cr treatment (0.5 g/kg/d) were initiated on Day 21 and lasted until Day 53. We determined the index of hyperresponsiveness, the serum levels of OVA-specific immunoglobulin (Ig)E and IgG(1), and the total and differential cell counts in bronchoalveolar lavage fluid. We also quantified airway inflammation, and the airway density of IL-4+, IL-5+, IL-2+, IFN-gamma+, and insulin-like growth factor (IGF)-1+ cells, collagen and elastic fibers, and airway smooth muscle thickness. Our results showed that creatine in OVA-sensitized mice increased hyperresponsiveness; eosinophilic inflammation; airway density of IL-4+, IL-5+, and IGF-1 inflammatory cells; airway collagen and elastin content; and smooth muscle thickness. The results show that creatine supplementation exacerbates the lung allergic response to OVA through a T helper cell type 2 pathway and increased IGF-1 expression.

Topics: Animals; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cell Count; Creatine; Dietary Supplements; Eosinophils; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Pneumonia; Respiratory Hypersensitivity; Respiratory System

The reason why particular inhaled Ags induce allergic sensitization while others lead to immune tolerance is unclear. Along with a genetic predisposition to atopy, intrinsic characteristics of these Ags must be important. A common characteristic of many allergens is that they either possess proteinase activity or are inhaled in particles rich in proteinases. Many allergens, such as house dust mite and cockroach allergens, have the potential to activate the proteinase-activated receptor (PAR)-2. In this study, we report that PAR-2 activation in the airways at the same time as exposure to inhaled Ags induces allergic sensitization, whereas exposure to Ag alone induces tolerance. BALB/c mice were administered OVA with a PAR-2 activating peptide intranasally. Upon allergen re-exposure mice developed airway inflammation and airway hyperresponsiveness, as well as OVA-specific T cells with a Th2 cytokine profile when restimulated with OVA in vitro. Conversely, mice given OVA alone or OVA with a PAR-2 control peptide developed tolerance. These tolerant mice did not develop airway inflammation or airway hyperresponsiveness, and developed OVA-specific T cells that secreted high levels of IL-10 when restimulated with OVA in vitro. Furthermore, pulmonary dendritic cell trafficking was altered in mice following intranasal PAR-2 activation. Finally, we showed that PAR-2-mediated allergic sensitization was TNF-dependent. Thus, PAR-2 activation in the airways could be a critical factor in the development of allergic sensitization following mucosal exposure to allergens with serine proteinase activity. Interfering with this pathway may prove to be useful for the prevention or treatment of allergic diseases.

Topics: Allergens; Animals; Cell Movement; Cytokines; Hypersensitivity; Immune Tolerance; Inhalation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptor, PAR-2; Th2 Cells; Tumor Necrosis Factor Inhibitors; Tumor Necrosis Factors

Allergic individuals rarely present with concurrent multiple-organ disease but, rather, with manifestations that privilege a specific site such as the lung, skin, or gastrointestinal tract. Whether the site of allergic sensitization influences the localization of Th2 immune-inflammatory responses and, ultimately, the organ-specific expression of disease, remains to be determined. In this study, we investigated whether both the site of initial Ag exposure and concomitant Th2 differentiation in specific lymph nodes (LNs) privileges Th2 memory responses to mucosal and nonmucosal sites, and whether this restriction is associated with a differential expression in tissue-specific homing molecules. In mice exposed to Ag (OVA) via the peritoneum, lung, or skin, we examined several local and distal LNs to determine the site of Ag-specific proliferation and Th2 differentiation. Whereas respiratory and cutaneous Ag exposure led to Ag-specific proliferation and Th2 differentiation exclusively in lung- and skin-draining LNs, respectively, Ag delivery to the peritoneum evoked responses in gut-associated, as well as distal thoracic, LNs. Importantly, only mice that underwent Th2 differentiation in thoracic- or gut-associated LNs mounted Th2 immune-inflammatory responses upon respiratory or gastric Ag challenge, respectively, whereas cutaneous Th2 recall responses were evoked irrespective of the site of initial sensitization. In addition, we observed the differential expression of gut homing molecules (CCR9, alpha(4), beta(7)) in gut-associated LNs and, unexpectedly, a universal induction of skin-related homing molecules (CCR4, CCR10) in all LNs. These data suggest that the site of initial Th2 differentiation and differential homing molecule expression restricts Th2 immune-inflammatory responses to mucosal, but not cutaneous, tissues.

Topics: Animals; Antigens; Cell Differentiation; Cytokines; Female; Gastrointestinal Tract; Hypersensitivity; Immunoglobulin E; Lung; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mucous Membrane; Ovalbumin; Peritoneum; Skin; Th2 Cells

Pathogenesis of allergic inflammatory disorders is characterized by allergen-induced IgE stimulated by Th2 cytokines including mainly IL-4 overproduction. To counteract IL-4 effects in sensitized-BALB/c mice, we prepared an IL-4 derivative immunogen, made of KLH and murine IL-4 heterocomplex, termed mIL-4 kinoid. Murine IL-4 kinoid immunized mice produced high titer of anti-IL-4 neutralizing Abs. In contrast to KLH control immunization kinoid immunization reversed the allergic IgE:IgG ratio hallmark in rBet v 1a sensitized mice and reduced pulmonary eosinophil recruitment and bronchial hyperreactivity in Ova-sensitized mice. These data pave the way to alternative therapies to combat allergic conditions.

Topics: Adjuvants, Immunologic; Allergens; Animals; Antigens, Plant; Eosinophils; Female; Hemocyanins; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Neutralization Tests; Ovalbumin; Recombinant Proteins

Vitamin A (VA) and its derivatives, the retinoids, are important factors for the development of the immune system. It has been shown in adult animals that proliferation of lymphocyte populations and antibody secretion are retinoid dependent, while little is known about the effects of retinoids during postnatal development. The aim of this study was to investigate the role of VA on allergic sensitisation during lactation and after weaning using an in vivo system for postnatal allergic sensitisation in mice. Different VA diets (basal/VA elimination/VA (as retinyl palmitate) supplemented) were fed to the dams throughout lactation and directly to the pups after weaning. Allergic sensitisation was induced with a single peritoneal ovalbumin (OVA) injection at day 28 after weaning. The phenotype of lymphocytes was analysed by flow cytometry and functional data were obtained by analysis of (IL-4/IFN-gamma) cytokine production and antibody production (OVA-specific IgG1 and IgE) in the offspring. VA/retinyl palmitate supplementation during lactation and after weaning decreased CD3+, CD4+, CD8+ and B220+ populations in splenic lymphocytes but also significantly enhanced IL-4 production and OVA-specific IgE after sensitisation. In contrast, mice fed VA-elimination diet displayed no significant alteration of lymphocyte numbers and a slightly increased IL-4 production. Our results showed that a single allergen injection during postnatal development induces allergic sensitisation whose degree is modified by the VA content of the maternal diet during lactation and the diet of the pups after weaning, indicating an important role of VA on the severity of the allergic sensitisation.

Topics: Allergens; Animals; Cytokines; Diet; Dietary Supplements; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lactation; Lymphocyte Count; Lymphocyte Subsets; Mice; Mice, Inbred BALB C; Ovalbumin; Phenotype; Spleen; Vitamin A; Weaning

We investigated the role of group V phospholipase A2 (gVPLA2) in OVA-induced inflammatory cell migration and airway hyperresponsiveness (AHR) in C57BL/6 mice. Repeated allergen challenge induced biosynthesis of gVPLA2 in airways. By aerosol, gVPLA2 caused dose-related increase in airway resistance in saline-treated mice; in allergic mice, gVPLA2 caused persistent airway narrowing. Neither group IIa phospholipase A2, a close homolog of gVPLA2, nor W31A, an inactive gVPLA2 mutant with reduced activity, caused airway narrowing in immune-sensitized mice. Pretreatment with MCL-3G1, a blocking Ab against gVPLA2, before OVA challenge blocked fully gVPLA2-induced cell migration and airway narrowing as marked by reduction of migrating leukocytes in bronchoalveolar lavage fluid and decreased airway resistance. We also assessed whether nonspecific AHR caused by methacholine challenge was elicited by gVPLA2 secreted from resident airway cells of immune-sensitized mice. MCL-3G1 also blocked methacholine-induced airway bronchoconstriction in allergic mice. Blockade of bronchoconstriction by MCL-3G1 was replicated in allergic pla2g5-/- mice, which lack the gene encoding gVPLA2. Bronchoconstriction caused by gVPLA2 in pla2g4-/- mice was comparable to that in pla2g4+/+ mice. Our data demonstrate that gVPLA2 is a critical messenger enzyme in the development of AHR and regulation of cell migration during immunosensitization by a pathway that is independent of group IVa phospholipase A2.

Topics: Animals; Antibodies; Antigens; Cell Movement; Gene Expression Regulation, Enzymologic; Hypersensitivity; Methacholine Chloride; Mice; Mice, Knockout; Ovalbumin; Phospholipases A; Phospholipases A2; Respiratory Hypersensitivity; Up-Regulation

Over the past several decades, there has been a significant increase in allergy and asthma in the world, which correlates with alterations in microflora and widespread use of antibiotics. The authors have developed a mouse model of antibiotics-induced microbiota disruption. In that model, mice were challenged by intranasal exposure to Aspergillus fumigatus allergens to explore the relation of allergic airway response and intestinal microflora disruption.. Sixty female BALB/c mice were divided at random into 6 groups with 10 mice in each. (1) First antibiotic therapy group: the mice were given oral cefoperazone for 7 days, on day 7, mice were inoculated with Candida albicans (10(9)/ml, 50 microl) orally. (2) First control group: the mice were treated as first antibiotic therapy group, but cefoperazone and Candida albicans were replaced by saline. The mice in groups (1) and (2) were sacrificed on day 8, and cecal contents were collected for quantitative analysis of the intestinal bacterial flora. (3) Antibiotic therapy and challenge group: the mice were treated as the first antibiotic therapy group, then challenged (day 9 and 16) by intranasal exposure to Aspergillus fumigatus allergen. (4) Second antibiotic therapy group: the mice were treated as the first antibiotic therapy group, then challenged (day 9 and 16) by intranasal exposure to saline. (5) Challenge group: the mice were treated as the first control group, then challenged (day 9 and 16) by intranasal exposure to Aspergillus fumigatus allergen. (6) Second control group: the mice were treated as the first control group, then challenged (day 9 and 16) by intranasal exposure to saline. The mice in (3) - (6) group were killed for analysis of allergic airway response on day 19.. The quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus in first antibiotic therapy group was significantly lower than that in the first control group, the quantity of Candida albicans increased in the first antibiotic therapy group as compared with the first control group. Mice intestinal microflora were disrupted with weight reduction and increased moisture in feces. After challenging with Aspergillus fumigatus allergens via intranasal inhalation, the total cell count, eosinophils, lymphocytes and neutrophils increased in BALF, especially in bronchoalveolar lavage fluid (BALF) from the mice in antibiotic therapy and challenge groups. IL-4 level in BALF from antibiotic therapy and challenge group (45.35 +/- 2.36) pg/ml was higher than that in the second control group (35.32 +/- 2.53) pg/ml. The expression of GATA-3 mRNA in the mice lung tissue (0.569 +/- 0.023) was higher than that in the second control group (0.410 +/- 0.020), and the ratios of T-bet/GATA-3 (0.578 +/- 0.021) decreased as compared with that in the second control group (0.804 +/- 0.035). IFN-gamma level in BALF from any group was not significantly different. In the absence of antibiotics, mice exposed to Aspergillus fumigatus allergen did not develop an allergic response in the airways.. The allergic (Th2) immune response can be induced by airway challenge with Aspergillus fumigatus allergen in the mice in which the intestinal microflora disruption resulted from antibiotic therapy, this result suggests that the intestinal microflora disruption resulted from antibiotic therapy is a risk factor for allergy and asthma.

Topics: Animals; Anti-Bacterial Agents; Antibiosis; Aspergillus fumigatus; Asthma; Bronchoalveolar Lavage Fluid; Cefoperazone; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Hypersensitivity, Immediate; Intestines; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory System

In this study, we examined the effects of dietary lactosucrose (LS, a non-digestible oligosaccharide) on the IgE response in mice immunized with ovalbumin (OVA)/alum. In addition to IgG1 and IgG2a responses, the anti-OVA IgE response in mice fed LS diets was dose-dependently suppressed, as compared with the control mice, while the serum total IgG levels were comparable. Moreover, dietary LS feeding inhibited antigen-specific IgE and IgG1 productions even after a second immunization. Regarding with cytokine production, when stimulated in vitro with OVA, splenocytes obtained from LS-fed mice produced a similar level of IFN-gamma, and lower levels of IL-4 and IL-5, as compared with the control mice. But IL-10 production by OVA-stimulated splenocytes was augmented in LS-fed mice, suggesting that IL-10 producing cells are responsible for the immunoregulatory effect of LS. Our findings indicate the further possibility that dietary LS supplementation can be used to prevent IgE-mediated allergic diseases.

Topics: Adjuvants, Immunologic; Allergens; Alum Compounds; Animals; Antibody Formation; Cytokines; Dietary Carbohydrates; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Immunosuppression Therapy; Mice; Ovalbumin; Spleen; Trisaccharides

Since B-1 cells were first described, their origin and function remain controversial. Given the ability to produce natural antibodies and large amounts of IL-10, there is a consensus about their role in innate immunity. More recently, however, B-1 cells have been associated to adaptive immunity as well, due to the demonstration of immunological memory and antigen presentation capability. Here we demonstrate that adoptive transfer of pre-sensitized B-1b cells (obtained from OVA-sensitized mice) to naïve B-1 deficient animals, drastically affects the ability of transplanted animals to mount an adaptive response upon immunization with OVA. In contrast to naïve B-1 populated mice, mice transplanted with sensitized B-1 exhibit lower anti-OVA antibody levels, milder footpad swelling in response to OVA subcutaneous injection and reduced granulomatous reaction to OVA-coated beads. Moreover, we show that these pre-sensitized B-1 cells, when acting as APCs, induce poor T cell proliferation in vitro when compared with macrophages or B-1 cells obtained from naïve mice. This property may be due in part to insufficient expression of the co-stimulatory molecule CD86, necessary for optimal antigen presentation. In conclusion, our data suggest a novel role for B-1 cells as part of suppressor mechanisms in the immune system.

Topics: Adoptive Transfer; Animals; B-Lymphocyte Subsets; Cell Proliferation; Hypersensitivity; Immune Tolerance; Immunity, Active; Immunity, Innate; Immunization; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Ovalbumin; T-Lymphocytes

The capacity of airway eosinophils, potentially pertinent to allergic diseases of the upper and lower airways, to function as professional APCs, those specifically able to elicit responses from unprimed, Ag-naive CD4(+) T cells has been uncertain. We investigated whether airway eosinophils are capable of initiating naive T cell responses in vivo. Eosinophils, isolated free of other APCs from the spleens of IL-5 transgenic mice, following culture with GM-CSF expressed MHC class II and the costimulatory proteins, CD40, CD80, and CD86. Eosinophils, incubated with OVA Ag in vitro, were instilled intratracheally into wild-type recipient mice that adoptively received i.v. infusions of OVA Ag-specific CD4(+) T cells from OVA TCR transgenic mice. OVA-exposed eosinophils elicited activation (CD69 expression), proliferation (BrdU incorporation), and IL-4, but not IFN-gamma, cytokine production by OVA-specific CD4(+) T cells in paratracheal lymph nodes (LN). Exposure of eosinophils to lysosomotropic NH(4)Cl, which inhibits Ag processing, blocked each of these eosinophil-mediated activation responses of CD4(+) T cells. By three-color fluorescence microscopy, OVA Ag-loaded eosinophil APCs were physically interacting with naive OVA-specific CD4(+) T cells in paratracheal LN after eosinophil airway instillation. Thus, recruited luminal airway eosinophils are distinct allergic "inflammatory" professional APCs able to activate primary CD4(+) T cell responses in regional LNs.

Topics: Adoptive Transfer; Allergens; Ammonium Chloride; Animals; Antigen Presentation; Antigen-Presenting Cells; Antigens, CD; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Eosinophils; Granulocyte-Macrophage Colony-Stimulating Factor; Histocompatibility Antigens Class II; Hypersensitivity; Inflammation; Infusions, Intravenous; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Transgenic; Ovalbumin; Th2 Cells; Up-Regulation

Artemisia princeps Pampanini (AP) was fermented with Bifidobacterium infantis K-525 and its antiasthmic effect investigated. AP and fermented AP (FAP) reduced the IgE level in the blood of ovalbumin-induced asthmic mice. Moreover, FAP reduced the IgE, proinflammatory cytokine IL-6, and IL-4 levels in the trachea, as well as in the lung of the experimental asthmic mice, whereas AP only reduced the IgE and IL-6 levels in the lungs. Nonetheless, AP and FAP both inhibited the mRNA expression of IL-6 and TNF-alpha in IgE-induced RBL-2H3 cells. The in vivo antiasthmic effect of FAP was more potent than that of AP. Therefore, these findings suggest that the enhanced antiasthmic effect of AP after bifidus fermentation was possibly due to the regulation of the proinflammatory cytokine biosynthesis of IL-6 and TNF-alpha.

Topics: Animals; Anti-Allergic Agents; Artemisia; Asthma; Bifidobacterium; Fermentation; Hypersensitivity; Immunoglobulin E; Interleukin-4; Interleukin-6; Lung; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Plant Extracts; Pruritus; RNA, Messenger

In addition to being an air pollutant, NO2 is a potent inflammatory oxidant generated endogenously by myeloperoxidase and eosinophil peroxidase. In these studies, we sought to determine the effects of NO2 exposure on mice with ongoing allergic airway disease pathology. Mice were sensitized and challenged with the antigen ovalbumin (OVA) to generate airway inflammation and subsequently exposed to 5 or 25 ppm NO2 for 3 days or 5 days followed by a 20-day recovery period. Whereas 5 ppm NO2 elicited no pathological changes, inhalation of 25 ppm NO2 alone induced acute lung injury, which peaked after 3 days and was characterized by increases in protein, LDH, and neutrophils recovered by BAL, as well as lesions within terminal bronchioles. Importantly, 25 ppm NO2 was also sufficient to cause AHR in mice, a cardinal feature of asthma. The inflammatory changes were ameliorated after 5 days of inhalation and completely resolved after 20 days of recovery after the 5-day inhalation. In contrast, in mice immunized and challenged with OVA, inhalation of 25 ppm NO2 caused a marked augmentation of eosinophilic inflammation and terminal bronchiolar lesions, which extended significantly into the alveoli. Moreover, 20 days postcessation of the 5-day 25 ppm NO2 inhalation regimen, eosinophilic and neutrophilic inflammation, pulmonary lesions, and AHR were still present in mice immunized and challenged with OVA. Collectively, these observations suggest an important role for NO2 in airway pathologies associated with asthma, both in modulation of degree and duration of inflammatory response, as well as in induction of AHR.

Topics: Animals; Bronchi; Bronchial Hyperreactivity; Dose-Response Relationship, Drug; Hypersensitivity; Mice; Mice, Inbred C57BL; Nitrogen Dioxide; Ovalbumin; Oxidants, Photochemical; Pneumonia

Antibody-antigen interactions in the airway initiate inflammation in acute asthma exacerbations. This inflammatory response is characterized by the recruitment of granulocytes into the airways. In murine models of asthma, granulocyte recruitment into the lung contributes to the development of airway hyperresponsiveness (AHR), mucus production, and airway remodeling. Leukotriene B4 is a mediator released following antigen challenge that has chemotactic activity for granulocytes, mediated through its receptor, BLT1. We investigated the role of BLT1 in granulocyte recruitment following antigen challenge. Wild-type mice and BLT1-/- mice were sensitized and challenged with ovalbumin (OVA) to induce acute allergic airway inflammation. In addition, to explore the relevance to antibody-antigen interactions, we injected OVA bound to anti-OVA IgG1 or anti-OVA IgE intratracheally into naïve wild-type and BLT1-/- mice. Cell composition of the lungs, cytokine levels, histology, and AHR were determined. After sensitization and challenge with ovalbumin, there was significantly reduced neutrophil and eosinophil recruitment into the airways of BLT1-/- mice compared with wild-type animals after one or two daily antigen challenges, but this difference was not seen after three or four daily antigen challenges. Mucus production and AHR were not affected. Intratracheal injection of OVA bound to IgG1 or IgE induced neutrophil recruitment into the airways in wild-type mice but not in the BLT1-/- mice. We conclude that BLT1 mediates early recruitment of granulocytes into the airway in response to antigen-antibody interactions in a murine model of acute asthma.

Topics: Animals; Antigen-Antibody Reactions; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemokines; Chemotaxis, Leukocyte; Disease Models, Animal; Granulocytes; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lung; Male; Mice; Mice, Knockout; Mucus; Neutrophils; Ovalbumin; Receptors, Leukotriene B4; Receptors, Purinergic P2

Soluble guanylyl cyclase (sGC) is an enzyme highly expressed in the lung that generates cGMP contributing to airway smooth muscle relaxation. To determine whether the bronchoconstriction observed in asthma is accompanied by changes in sGC expression, we used a well-established murine model of allergic asthma. Histological and biochemical analyses confirmed the presence of inflammation in the lungs of mice sensitized and challenged with ovalbumin (OVA). Moreover, mice sensitized and challenged with OVA exhibited airway hyperreactivity to methacholine inhalation. Steady-state mRNA levels for all sGC subunits (alpha1, alpha2, and beta1) were reduced in the lungs of mice with allergic asthma by 60-80%, as estimated by real-time PCR. These changes in mRNA were paralleled by changes at the protein level: alpha1, alpha2, and beta1 expression was reduced by 50-80% as determined by Western blotting. Reduced alpha1 and beta1 expression in bronchial smooth muscle cells was demonstrated by immunohistochemistry. To study if sGC inhibition mimics the airway hyperreactivity seen in asthma, we treated naïve mice with a selective sGC inhibitor. Indeed, in mice receiving ODQ the methacholine dose response was shifted to the left. We conclude that sGC expression is reduced in experimental asthma contributing to the observed airway hyperreactivity.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Enzyme Inhibitors; Guanylate Cyclase; Homeostasis; Hypersensitivity; Isoenzymes; Methacholine Chloride; Mice; Ovalbumin; Oxadiazoles; Quinoxalines; Receptors, Cytoplasmic and Nuclear; RNA, Messenger; Soluble Guanylyl Cyclase

Modern persistent organic pollutants (POPs) contamination are logical candidates in the investigation of the, as yet, unexplained association between allergic disease and progressive industrialisation. POPs have been detected in human cord blood, placental tissues and breast milk, and the reported association between cord blood IgE levels and cord/placental POP levels has raised concerns about potential immunological effects in early life.. The initial aim of this study was to determine if POPs were detectable in maternal blood, cord blood, placental tissues, adipose tissue and breast milk samples from randomly selected Western Australian women (n = 31), where allergic disease is epidemic. Gas chromatography was used to detect polychlorinated biphenyl compounds [PCBs] (as Aroclor 1232, 1254, 1260) and organochlorine (OC) pesticides, including p,p'-DDT, p,p'-DDE, hexachlorobenzene (HCB), lindane, heptachlor epoxide, dieldrin and chlordane. Secondly, we assessed the relationship between POP levels detected in vivo and maternal and neonatal responses (cytokine and lymphoproliferation) to allergens and mitogens.. Low level POP contamination was detected in adipose tissue and breast milk (but not in cord blood, maternal blood or placental tissues). The most ubiquitous compound found in over 90% of adipose tissues samples was a OC metabolite of DDT, p,p'-DDE (median 0.07 mg/kg; interquartile range [IQR] 0.05-0.12). However, the majority of other OC compounds were not detectable and PCB were not detectable in any samples. The three main residues detected in breast milk were p,p'-DDE (0.003 mg/l; 0.001-0.009), dieldrin (0.001 mg/l; 0.001-0.046) and HCB (0.001 mg/l; 0.001-0.001). These levels are significantly lower than reported over 20 years ago. There were no consistent relationships between POP levels in vivo and maternal or infant responses, with the exception of a significant inverse association (Spearman rank correlation: r = -0.406, p = 0.049) between maternal adipose tissue levels of OC p,p'-DDE and maternal T helper cell Type 1 interferon [IFN] gamma to mitogens.. This study provides the first evidence (in Australia) since the early 1990's that adipose OC levels have continued to fall. The negligible levels in this randomly selected group are significantly lower than those previously recorded, suggesting that POP contamination (at biologically relevant levels) is not likely to be a major contributing factor in the increasing rates of allergy in Western Australia. However, the relationship between Th1 immune function and OC contamination is consistent with other reports and is worth investigating as a relevant factor in populations where OC contamination is greater.

Topics: Adipose Tissue; Adolescent; Adult; Allergens; Cytokines; Environmental Monitoring; Environmental Pollutants; Female; Fetal Blood; Humans; Hydrocarbons, Chlorinated; Hypersensitivity; Infant, Newborn; Leukocytes, Mononuclear; Maternal Exposure; Milk, Human; Ovalbumin; Placenta; Pregnancy; Pyroglyphidae; Western Australia

Hardwood smoke (HWS) from wood burning stoves and fireplaces can be a significant contributor to the composition of ambient air pollution. We hypothesize that the inhalation of HWS by ovalbumin (OVA)-sensitized mice with preexisting lung inflammation leads to the exacerbation of allergic airway responses. Two different models were employed to characterize the effects of inhaled wood smoke on allergic airway inflammation. In both models, male BALB/c mice were sensitized by injection with OVA and alum. In one model, mice were challenged by inhalation with OVA 1 day prior to exposure to HWS (30, 100, 300, or 1000 microg particulate matter [PM]/m(3)) for 6 h/day on 3 consecutive days. In the other model, mice were exposed by inhalation to OVA, rested for 11 days, were exposed to HWS for 3 consecutive days, and then were exposed to OVA immediately after the final HWS exposure. Bronchoalveolar lavage (BAL), and blood collection were performed approximately 18 h after the last HWS or OVA exposure. HWS exposure after the final allergen challenge (first model) led to a significant increase in BAL eosinophils only at the 300 microg/m(3) level. In contrast, changes in BAL cells did not reach statistical significance in the second model. There were no HWS-induced changes in BAL interleukin (IL)-2, IL-4, IL-13, and interferon (IFN)gamma levels in either model following OVA challenge. These results suggest that acute HWS exposure can minimally exacerbate some indices of allergic airway inflammation when a final OVA challenge precedes HWS exposure, but does not alter Th1/Th2 cytokine levels.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cytokines; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Inhalation Exposure; Lung Diseases; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Smoke; Wood

At present there are conflicting results from studies investigating the role of corticosteroids in inhibiting airway remodeling in asthma. We have used a mouse model to determine whether administration of corticosteroids prevents the development of allergen-induced structural features of airway remodeling. Mice treated with corticosteroids were subjected to repetitive ovalbumin (OVA) challenge for 3 mo, at which time levels of peribronchial fibrosis and the thickness of the peribronchial smooth muscle layer were assessed by immunohistology, levels of transforming growth factor (TGF)-beta1 by ELISA, and the number of alpha-smooth muscle actin+/Col-1+ peribronchial myofibroblasts by immunohistochemistry. Corticosteroids significantly reduced allergen-induced increases in peribronchial collagen deposition and levels of total lung collagen but did not reduce allergen-induced increases in the thickness of the peribronchial smooth muscle layer. Levels of lung TGF-beta1 were significantly reduced in mice treated with systemic corticosteroids, and this was associated with a significant decrease in the number of peribronchial inflammatory cells that expressed TGF-beta1, including eosinophils and mononuclear cells. Corticosteroids also significantly reduced the number of peribronchial myofibroblasts. Overall, these studies demonstrate that administration of corticosteroids significantly reduces levels of allergen-induced peribronchial fibrosis. The reduction in peribronchial fibrosis mediated by corticosteroids is likely to be due to several mechanisms including inhibition of expression of TGF-beta1, a reduction in the number of peribronchial inflammatory cells expressing TGF-beta1 (eosinophils, macrophages), as well as by corticosteroids reducing the accumulation of peribronchial myofibroblasts that contribute to collagen expression.

Topics: Actins; Adrenal Cortex Hormones; Animals; Bronchi; Bronchitis; Collagen; Fibroblasts; Fibronectins; Hypersensitivity; Immunologic Techniques; Lung; Mice; Mice, Inbred BALB C; Mucus; Muscle, Smooth; Myocytes, Smooth Muscle; Ovalbumin; Staining and Labeling; Transforming Growth Factor beta; Transforming Growth Factor beta1

Steroid treatment of allergic eosinophilic airway diseases is considered to attenuate cell recruitment by inhibiting several chemokines and to cause eosinophil clearance through inducement of apoptosis of these cells. However, roles of these mechanisms in the actions of steroids in vivo have not been fully established. Also, as regards clearance of tissue eosinophils other mechanisms than apoptosis may operate in vivo.. This study explores anti-inflammatory effects of steroids instituted during either development or resolution of airway allergic inflammation.. Immunized mice were subjected to week-long daily allergen challenges (ovalbumin). Steroid treatment was instituted either amidst the challenges or exclusively post-allergen challenge. CC chemokines, goblet cell hyperplasia, occurrence of eosinophil apoptosis, and airway tissue as well as lumen eosinophilia were examined at different time-points.. Daily steroids instituted amid the allergen challenges non-selectively attenuated a range of chemokines, permitted egression of tissue eosinophils into airway lumen to increase, and reduced development of lung tissue eosinophilia. Steroid treatment instituted post-challenge selectively inhibited the CC-chemokine regulation upon activation, normal T cell expressed and secrted (RANTES), permitted continued egression of eosinophils into airway lumen, and resolved the tissue eosinophilia. Eosinophil apoptosis rarely occurred at development and resolution of the allergic eosinophilic inflammation whether the animals were steroid treated or not. However, anti-Fas monoclonal antibodies given to mice with established eosinophilia post-challenge produced apoptosis of the tissue eosinophils indicating that apoptotic eosinophils, if they occur, are well detectible in vivo.. Airway tissue eosinophils are likely eliminated through egression into airway lumen with little involvement of apoptosis and phagocytosis. Our data further suggest that therapeutic steroids may resolve airway inflammation by permitting clearance of tissue eosinophils through egression and inhibiting RANTES-dependent cell recruitment to lung tissues.

Topics: Allergens; Animals; Apoptosis; Biomarkers; Bronchoalveolar Lavage Fluid; Budesonide; Chemokine CCL11; Chemokine CCL4; Chemokine CCL5; Chemokines, CC; Eosinophils; Epithelial Cells; Glucocorticoids; Hypersensitivity; Interleukin-5; Lung; Macrophage Inflammatory Proteins; Male; Mice; Mice, Inbred C57BL; Models, Animal; Ovalbumin; RNA, Messenger; Staining and Labeling

The present study reports the anti-allergic activity of a group of six different tetranortriterpenoids (TNTP) isolated from the seeds of Carapa guianensis Aublet: 6a-acetoxygedunin, 7-deacetoxy-7-oxogedunin, andirobin, methyl angolensate, 6a-acetoxyepoxyazadiradione and gedunin. Oral pretreatment with TNTP significantly inhibited total leukocyte and eosinophil accumulation in C57BL/10 mice pleural cavities 24 h after the intrathoracic (i.t.) injection of ovalbumin (OVA), but had no effect on CD4, CD8 or gammadelta T lymphocyte accumulation. Pleural washes recovered from 6 h OVA-stimulated mice (OPW) pretreated with TNTP failed to induce shape change in eosinophil in vitro, indicating the inhibition of eosinophilotactic chemokines by TNTP. In accordance with such results, ELISA assays showed decreased levels of CCL11/eotaxin and IL-5 in OPW recovered from TNTP pretreated mice within 6 h. TNTP oral pretreatment inhibited nuclear factor-kappaB (NFkappaB) translocation into the nucleus in pleural leukocytes recovered from previously sensitized mice after antigenic challenge. In addition, the incubation of splenocytes recovered from previously sensitized mice with TNTP also inhibited NFkappaB activation after OVA stimulation. Taken together, these results indicate that the inhibition of allergic eosinophilia by TNTP is correlated with the inhibition of CCL11/eotaxin and IL-5 generation through NFkappaB signaling pathway impairment in mice.

Topics: Allergens; Animals; Blotting, Western; Cell Nucleus; Cell Shape; Chemokine CCL11; Chemokines, CC; Enzyme-Linked Immunosorbent Assay; Eosinophils; Flow Cytometry; Hypersensitivity; Immunoblotting; Immunohistochemistry; Interleukin-5; Leukocytes; Mice; Mice, Inbred C57BL; Neutrophils; NF-kappa B; Ovalbumin; Pleurisy; Subcellular Fractions; Triterpenes

Ganoderma tsugae (a Chinese mushroom Songshan lingzhi) cultivated in Taiwan is extensively used in Chinese traditional medicine to treat different diseases. To determine whether G. tsugae has anti-inflammatory effects on bronchoalveolar inflammation in vivo, we investigated the anti-inflammatory effects of G. tsugae products, YK01 and YK07, on bronchoalveolar inflammation using an airway sensitization and challenge mouse model. Female BALB/c mice were weekly sensitized by intraperitoneal injection of ovalbumin (OVA) three times and challenged with aerosolized OVA twice. Differential cell counts of infiltrating leukocytes, inflammatory mediators, cytokines in bronchoalvelor lavage fluid (BALF) of OVA-challenged mice were examined after continuously consuming G. tsugae diets for 5 weeks. We found that supplementation of G. tsugae significantly decreased total infiltrating leukocytes and lymphocyte percentage in BALF in the experimental groups. Supplementation of G. tsugae also significantly reduced inflammatory mediators in BALF including histamine, prostaglandin E2, eotaxin, and protein levels, however the levels of pro-inflammatory cytokines, interleukin (IL)-1beta and IL-6, in BALF did not significantly change. These results suggest that both G. tsugae supplementation diets YK01 and YK07 might alleviate bronchoalveolar inflammation via decreasing the infiltration of inflammatory cells and the secretion of inflammatory mediators into the local tissues of lungs and airways. Further, these results indicate that the relief of bronchoalveolar inflammation in an airway sensitization murine model provides a possible therapeutic application for G. tsugae in allergic asthma.

Topics: Alveolitis, Extrinsic Allergic; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Count; Chemokine CCL11; Chemokines; Chemokines, CC; Cytokines; Dinoprostone; Enzyme-Linked Immunosorbent Assay; Female; Ganoderma; Histamine Release; Hypersensitivity; Mice; Mice, Inbred BALB C; Mycelium; Nitric Oxide; Ovalbumin; Proteins; Th1 Cells; Th2 Cells

Using guinea pig tracheal preparations, we have recently shown that endogenous arginase activity attenuates inhibitory nonadrenergic noncholinergic (iNANC) nerve-mediated airway smooth muscle relaxation by reducing nitric oxide (NO) production--due to competition with neuronal NO-synthase (nNOS) for the common substrate, L-arginine. Furthermore, in a guinea pig model of allergic asthma, airway arginase activity is markedly increased after the early asthmatic reaction (EAR), leading to deficiency of agonist-induced, epithelium-derived NO and subsequent airway hyperreactivity. In this study, we investigated whether increased arginase activity after the EAR affects iNANC nerve-derived NO production and airway smooth muscle relaxation.. Electrical field stimulation (EFS; 150 mA, 4 ms, 4 s, 0.5-16 Hz)-induced relaxation was measured in tracheal open-ring preparations precontracted to 30% with histamine in the presence of 1 microM atropine and 3 microM indomethacin. The contribution of NO to EFS-induced relaxation was assessed by the nonselective NOS inhibitor Nomega-nitro-L-arginine (L-NNA, 100 microM), while the involvement of arginase activity in the regulation of EFS-induced NO production and relaxation was investigated by the effect of the specific arginase inhibitor Nomega-hydroxy-nor-L-arginine (nor-NOHA, 10 microM). Furthermore, the role of substrate availability to nNOS was measured in the presence of exogenous L-arginine (5.0 mM).. At 6 h after ovalbumin-challenge (after the EAR), EFS-induced relaxation (ranging from 3.2 +/- 1.1% at 0.5 Hz to 58.5 +/- 2.2% at 16 Hz) was significantly decreased compared to unchallenged controls (7.1 +/- 0.8% to 75.8 +/- 0.7%; P < 0.05 all). In contrast to unchallenged controls, the NOS inhibitor L-NNA did not affect EFS-induced relaxation after allergen challenge, indicating that NO deficiency underlies the impaired relaxation. Remarkably, the specific arginase inhibitor nor-NOHA normalized the impaired relaxation to unchallenged control (P < 0.05 all), which effect was inhibited by L-NNA (P < 0.01 all). Moreover, the effect of nor-NOHA was mimicked by exogenous L-arginine.. The results clearly demonstrate that increased arginase activity after the allergen-induced EAR contributes to a deficiency of iNANC nerve-derived NO and decreased airway smooth muscle relaxation, presumably via increased substrate competition with nNOS.

Topics: Animals; Arginase; Arginine; Asthma; Bronchoconstriction; Electric Stimulation; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Male; Muscle Relaxation; Muscle, Smooth; Nitric Oxide; Nitric Oxide Synthase Type I; Ovalbumin; Specific Pathogen-Free Organisms; Trachea

Peripheral tolerance can be induced after the feeding of Ag, which is referred to as oral tolerance. We demonstrated in this study that the oral administration of OVA induced tolerance in an experimental model of asthma in mice, and investigated which cells function as the regulatory cells in the transfer of this oral tolerance. In OVA-fed mice, the percentage of eosinophils in bronchoalveolar lavage fluid, serum IgE levels, airway hyperresponsiveness, and mRNA levels of IL-13 and eotaxin were significantly lower than found in nonfed mice. Histological examination of lung tissue showed a suppression of the accumulation of inflammatory cells in the peribronchial area of OVA-fed mice. Feeding after the first immunization or between the first and the second immunization suppressed these findings, whereas feeding just before the airway Ag challenge did not. The suppression of disease in OVA-fed mice was successfully transferred by injection of whole spleen cells of OVA-fed mice. When CD11c+ dendritic cells (DCs) were removed from splenocytes, this transfer of suppression was completely abolished. The injection of splenic DCs purified from OVA-fed mice alone transferred the suppression, whereas the injection of splenic DCs from naive mice that were cocultured with OVA in vitro did not. These data suggest that not only CD4+ T cells, but also CD11c+ DCs induced by Ag feeding are important for the active transfer of oral tolerance in this murine experimental model of asthma.

Topics: Administration, Oral; Adoptive Transfer; Animals; Antigens; Asthma; CD11c Antigen; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Dose-Response Relationship, Immunologic; Eosinophilia; Hypersensitivity; Immune Tolerance; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Spleen

Bronchial asthma is defined as a chronic airway inflammatory disease characterized by sustained activation of many inflammatory cells including mast cells. Urocortin (UCN) is synthesized and secreted by human mast cells and activated mast cells release more UCN. On the other hand, UCN can induce mast cell degranulation and generation of many proinflammatory factors. The purpose of this study was to examine the expression profile of UCN in rat lung with allergic asthma. Twenty-four male Sprague-Dawley rats were allocated to normal control, asthma model, and dexamethasone group, respectively. Animals were actively sensitized by subcutaneous injection of ovalbumin (OVA) and challenged by an aerosol of 1% OVA 2 weeks after sensitization. Both UCN mRNA and peptide were expressed in normal rat lungs. Rats in asthma model group developed severe infiltration of inflammatory cells and inflammation in airway, together with a significantly up-regulated expression of urocortin mRNA detected by semi-quantitative reverse transcriptase-polymerase chain reaction and peptide measured both by immunohistochemistry and Western blot analysis. In contrast, treatment with dexamethasone resulted in markedly ameliorated airway inflammation and alleviated airway inflammatory cell infiltration, coupled with a significantly decreased urocortin expression. Regression analysis revealed a positive correlation between urocortin expression and the number of inflammatory cells in bronchoalveolar lavage fluid (P<0.01). In the present study, we first demonstrated that UCN was locally produced in rat lungs and expressed more pronouncedly in inflammatory airway of asthmatic rats. Glucocorticoid treatment markedly reduced the production of UCN in asthmatic lung tissues. Peripherally produced UCN in lung may act as a possible local autocrine and paracrine immune-inflammatory mediator in inflammatory airway of allergic asthma rats.

Topics: Actins; Animals; Asthma; Blotting, Western; Bronchi; Bronchoalveolar Lavage; Corticotropin-Releasing Hormone; Dexamethasone; Disease Models, Animal; Epithelium; Gene Expression Regulation; Glucocorticoids; Hypersensitivity; Immunohistochemistry; Inflammation; Lung; Male; Mast Cells; Models, Statistical; Ovalbumin; Peptides; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Time Factors; Up-Regulation; Urocortins

Eotaxin is a chemokine that has high potency and selectivity as a chemoattractant agent for eosinophils, signalling exclusively through the CCR3 receptor. Eotaxin is upregulated in the lungs within 3 h of antigen challenge, levels peak at 6 h in lung tissue and bronchoalveolar (BAL) fluid, and fall within 12 h of exposure. This study aimed to look at the effect(s) of eotaxin inhalation on airway function in guinea pigs, to determine if the expected inflammatory cell (eosinophil) infiltration could induce airway hyperreactivity (AHR) and a bronchoconstrictor response equivalent to the late asthmatic response (LAR) seen after antigen challenge. Animals were sensitised with 100 microg/ml OA with a dose on days 1 and 5. Airway responses to inhaled eotaxin (10 or 20 microg/ml) were determined by whole body plethysmography as the change in specific airway conductance (sGaw). Inhaled histamine (1mM) was used to investigate AHR, and cell influx was determined by BAL. Senitised animals exposed to 10 microg/ml eotaxin did not reveal a bronchoconstrictor response or AHR and cellular infiltration to the lungs was not evident 24 h after exposure. Both sensitised and non-sensitised animals exposed to 20 microg/ml eotaxin however revealed a significant bronchoconstrictor response 6h post-challenge, with reductions in sGaw of -27.0+/-6.6% and -32.3+/-6.8%, respectively. Both groups also displayed a bronchoconstrictor response to inhaled histamine 24h after exposure, indicating AHR, and a significant increase in both total and differential cell counts. Sensitised animals, however, revealed a significant increase in cell influx compared to non-sensitised animals. Nebulised eotaxin can reveal a LAR, AHR to inhaled histamine, and cellular infiltration to the lungs, possibly via the mobilisation of eosinophils from the bone marrow, and their subsequent recruitment to the airways.

Topics: Administration, Inhalation; Airway Resistance; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cell Count; Chemokine CCL11; Chemokines, CC; Data Interpretation, Statistical; Guinea Pigs; Histamine; Hypersensitivity; Inflammation; Male; Ovalbumin; Plethysmography, Whole Body; Respiratory Physiological Phenomena

Oral tolerance is a T-cell mediated phenomenon defined by inhibition of immune responsiveness to a protein previously contacted by the oral route. Oral tolerance may prevent autoimmune and allergic diseases that involve the recruitment and/or activation of different cell types including mast cells, neutrophils, eosinophils, monocytes and lymphocytes. The mechanisms by which oral tolerance avoids these immunological disorders are still controversial. Herein we used a murine model of ovalbumin (OVA)-induced peritonitis to investigate the effect of oral tolerance on allergic inflammation. Frequency of leucocyte subpopulations was evaluated by global and differential cell counts in peritoneal lavage fluid, peripheral blood, and bone marrow. Changes on lymphocyte subsets and adhesion molecules expression by these cells were analysed by flow cytometry. As compared with OVA-immune mice, intraperitoneal challenge of tolerant animals with OVA resulted in a significantly milder peritonitis, mostly affecting neutrophils and eosinophils; a concomitant reduction in total white blood cell counts was also observed, mainly because of lower neutrophil and eosinophil counts. Eosinophils, but not neutrophils, were also reduced in the bone-marrow of OVA-challenged tolerant mice. No changes occurred in total peritoneal lymphocyte counts in OVA-tolerant mice, however, there was a significant decrease in CD3+ CD8+ T cells and an increase in B cells (CD45R+) in these animals as compared to immune OVA-challenged animals. Altered expression of CD18 and CD54, respectively, in blood and peritoneal lymphocytes was also noted. These results suggest that, in addition to local specific effects, oral tolerance has systemic effects on the mobilization of leucocytes and bone-marrow eosinopoiesis.

Topics: Animals; Antigens; Ascitic Fluid; Bone Marrow; Cell Adhesion Molecules; Eosinophilia; Eosinophils; Female; Granulocytes; Hypersensitivity; Immune Tolerance; Leukocyte Count; Lymphocyte Subsets; Mice; Mice, Inbred Strains; Neutrophils; Ovalbumin; Peritonitis

The mammalian chitinase family includes members both with and without enzymatic activity against chitin, a product of fungal cell walls, exoskeletons of crustaceans and insects, and the microfilarial sheaths of parasitic nematodes. Two members of that family, Ym1 and acidic mammalian chitinase (AMCase), are strongly upregulated in pulmonary T helper (Th) 2 inflammation but not in Th1 inflammation. The sites of expression of these products are incompletely known. We show here that, in two different models of Th2 inflammation, Ym1 and AMCase are mutually exclusively expressed in proximal vs. distal airway epithelium, respectively, whereas both are expressed in alveolar macrophages. This regional difference along the airway corresponds to the previously noted distinction between mucus positive proximal cells and mucus negative distal cells under the same conditions. Among distal cells, AMCase colocalizes with epithelial cells expressing the Clara cell marker Clara cell secretory protein. These AMCase-expressing cells retain expression of FOXA2, a transcription factor whose downregulation in association with IL-13 signaling has previously been associated with production of mucus in proximal airway epithelial cells. These results provide evidence that secretory cells of proximal and distal airways undergo fundamentally different gene expression programs in response to allergic inflammation. Furthermore, AMCase provides the first positive molecular marker of distal Clara cell secretory protein-expressing cells under these conditions.

Topics: Animals; Biomarkers; Chitinases; Hepatocyte Nuclear Factor 3-beta; Hypersensitivity; Inflammation; Interleukin-13; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Respiratory Mucosa; Uteroglobin

A 95% ethanol extract from whole aerial parts of Euphorbia hirta (EH A001) showed antihistaminic, antiinflammatory and immunosuppressive properties in various animal models. EH A001 inhibited rat peritoneal mast cell degranulation triggered by compound 48/80. It significantly inhibited dextran-induced rat paw edema. EH A001 prevented eosinophil accumulation and eosinophil peroxidase activity and reduced the protein content in bronchoalveolar lavage fluid (BALF) in a 'mild' model of asthma. Moreover, the CD4/CD8 ratio in peripheral blood was suppressed. EH A001 attenuated the release of interleukin-4 (IL-4) and augmented interferon-gamma (IFN-gamma) in ovalbumin-sensitized mouse splenocytes. The results were compared with the effects of known compounds, ketotifen, cetirizine and cyclophosphamide. These findings demonstrated that Euphorbia hirta possessed significant activity to prevent early and late phase allergic reactions.

Topics: Animals; Bronchoalveolar Lavage Fluid; CD4-CD8 Ratio; Cytokines; Edema; Eosinophil Peroxidase; Eosinophils; Euphorbia; Histamine Antagonists; Hypersensitivity; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Rats; Rats, Wistar; Spleen

Chondroitin sulfate (CS) was administered orally to BALB/c mice immunized intraperitoneally with ovalbumin (OVA) and/or dinitrophenylated OVA. The titers of antigen-specific IgE and IgG1 in mouse sera were determined. The antigen-specific IgE production by mice fed ad libitum with CS was significantly inhibited. We also examined the effect of feeding CS on immediate-type hypersensitivity. One hour after antigen stimulation, the ears of mice fed with CS swelled less than those of the control mice. Furthermore, the rise in serum histamine in the mice fed with CS under active systemic anaphylaxis was significantly lower than that in the controls. We next examined the pattern of cytokine production by splenocytes from mice followed by re-stimulation with OVA in vitro. The splenocytes from the mice fed with CS produced less interleukin (IL)-5, IL-10, and IL-13 than those from the control group. In contrast, the production of interferon-gamma and IL-2 by the splenocytes of mice fed with CS was not significantly different from those in the control mice. In addition, the production of transforming growth factor-beta from the splenocytes of mice fed with CS was significantly higher than that of the control mice. Furthermore, we showed that the percentages of CD4(+) cells, CD8(+) cells, and CD4(+)CD25(+) cells in the splenocytes of mice fed with CS are significantly higher than those of the control. These findings suggest that oral intake of CS inhibits the specific IgE production and antigen-induced anaphylactic response by up-regulating regulatory T-cell differentiation, followed by down-regulating the Th2 response.

Topics: Administration, Oral; Animals; Chondroitin Sulfates; Ear; Edema; Hypersensitivity; Immunoglobulin E; Magnetic Resonance Spectroscopy; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Quillaja saponin is the extract from the balk of a South American tree, and it is considered to modulate immunological responses. We hypothesized that Quillaja saponin may change allergy-associated cytokine profile and antigen-specific immune responses. The purpose of this study is to investigate whether Quillaja saponin can suppress ovalbumin (OVA)-induced IgE-mediated allergic responses through promoting a dominant Th1 immune response. The spleen cells from BALB/c mice, which were primed by OVA, were used for an in vitro challenge test. The level of total and OVA-specific IgE, IL-4, IFN-gamma, and IL-12 was determined by enzyme-linked immunosorbent assay (ELISA). BALB/c mice were orally administered with saponin for 35 days. The mice were immunized intraperitoneally with OVA on days 14 and 21. After intraperitoneal challenge with OVA on day 35, anaphylactic symptoms were monitored. Total and specific IgE and IgG, specific IgG1 and IgG2a, and histamine levels in serum were analyzed by ELISA. The increase of IL-12 and IFN-gamma levels was observed in the presence of Quillaja saponin, while the IL-4 level was decreased. Furthermore, Quillaja saponin suppressed total and OVA-specific IgE secretion in spleen cells. Balb/c mice that were orally administered Quillaja saponin exhibited lower total and OVA-specific IgE and OVA-specific IgG secretions, whereas total IgG levels remained unchanged. Suppression of OVA-specific IgG1 and an increase of OVA-IgG2a were observed in mice fed saponin. Quillaja saponin also decreased serum histamine levels and diminished anaphylactic symptoms. The present study indicates that Quillajasaponin can suppress allergen-specific IgE-mediated reactivity in a murine model of food allergy, which results from shifting from a Th2-dominated to a Th1-dominated immune response.

Topics: Animals; Female; Hypersensitivity; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Quillaja; Th1 Cells; Th2 Cells

The prevalence of atopic disease among children in the formerly socialist countries in Europe, with a life style similar to that prevailing in Western Europe 30-40 years ago, is low, whereas there has been a pronounced increase in industrialized countries over the last decades. The environment during infancy influences the risk of developing allergy for many years, perhaps even for life.. To investigate the development of allergen-specific cytokine responses during the first 2 years of life in two geographically adjacent countries with marked differences in living conditions and incidence of atopic diseases, i.e. Estonia and Sweden.. The development of immune responses to food (beta-lactoglobulin (BLG) and ovalbumin (OVA)) and inhalant (cat and birch) allergens was studied from birth up to the age of 2 years in 30 Estonian and 76 Swedish infants. Clinical investigation and skin prick tests were performed and blood samples were obtained at birth and at 3, 6, 12 and 24 months.. The levels of IL-5, IL-10 and IL-13 secreted by peripheral blood mononuclear cells stimulated with BLG, OVA and cat allergen in Estonian and Swedish infants declined during the first 3 months of life. All cytokines then progressively increased in the Swedish infants, indicating the replacement of non-specifically responding immature cord blood T cells with specific T memory cells, which are primed postnatally. The resurgence of allergen-specific responses in the Estonian infants was less marked. These differences were particularly notable for birch-specific T cell responses, which correlated with development of atopic disease in the Swedish children.. The development of specific T cell memory to food and inhalant allergens during the first 2 years of life differs between infants living in Sweden and Estonia, and mirrors the disparate patterns of expression of allergic disease which subsequently develops in the respective populations.

Topics: Allergens; Animals; Betula; Cats; Cells, Cultured; Cytokines; Estonia; Humans; Hypersensitivity; Infant; Interferon-gamma; Interleukin-10; Interleukin-13; Interleukin-4; Interleukin-5; Interleukin-9; Lactoglobulins; Leukocytes, Mononuclear; Life Style; Ovalbumin; Sweden; T-Lymphocytes

The relationship between LPS exposure and allergic asthma is poorly understood. Epidemiologic studies in humans have found that exposure to LPS can protect, have no effect, or exacerbate allergic asthma. Similarly, LPS has had variable effects on allergic pulmonary inflammation in the mouse, depending on the model used. In the present study, we studied the effect of very low doses of LPS in models of both short-term and long-term allergen challenge. When challenged with allergen for short periods, wild-type and tlr4-deficient mice had similar responses. However, when challenged for periods of 1 wk or longer, tlr4-deficient mice developed dramatically increased airway eosinophils, serum IgE, and Th2 cytokines compared with similarly challenged, genetically matched C57BL/6 mice. The relative attenuation of allergic responses seen in C57BL/6 mice was dependent on bone marrow-derived cell-specific expression of tlr4, and was not associated with an increase in Th1 responses. The number of dendritic cells in lungs of challenged tlr4-deficient mice was significantly increased compared with those in challenged C57BL/6 mice. No differences were seen in the abilities of naive C57BL/6 and tlr4-deficient mice to develop allergen-specific tolerance after exposure to similar preparations of OVA, suggesting that tolerance and regulation of existing inflammation develop through different mechanisms. The attenuation of eosinophilic inflammation in C57BL/6 mice was abolished when these mice were challenged with OVA supplemented with additional LPS. Together, these findings show that low doses of endotoxin can have regulatory effects on allergic inflammation, particularly in the setting of ongoing allergen exposure.

Topics: Allergens; Animals; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Signal Transduction; Toll-Like Receptor 2; Toll-Like Receptor 4

Dendritic cells are the most powerful of the antigen-presenting cells and are known to play important roles in sensitization and inflammation in allergen-specific asthma. Various cytokines and chemokines are involved in the maturation and activation of dendritic cells. Among them is CC chemokine ligand (CCL)21, a key chemokine in the entry of naive T cells and antigen-stimulated dendritic cells into the T-cell zones of secondary lymphoid organs, which is a critical process in antigen-specific T-cell activation.. We studied the role of CCL21 in airway inflammation in asthma by using BALB/c-plt/plt (plt) mice, which possess genetic defects in expression of both CCL21 and CCL19.. Plt and control BALB/c mice were immunized with ovalbumin and alum 4 times and thereafter were subjected to a 2-week regimen of ovalbumin inhalation.. In plt mice, ovalbumin-specific IgE response was delayed compared with control BALB/c mice, but they had the same level of response after final immunization. Although airway inflammation and response to acetylcholine were significantly reduced compared with BALB/c mice, significant eosinophilic inflammation and hyperresponsiveness were also observed in plt mice after 2 weeks of inhalation. Four weeks after cessation of inhalation, airway inflammation and hyperresponsiveness in plt mice were greater than in BALB/c mice. At the time of resolution of airway inflammation, IL-10 production was enhanced in BALB/c mice but not in plt mice.. The chemokines CCL21 and CCL19 were critical for resolution of airway inflammation.. The findings about the chemokines for induction and resolution of inflammation are key to establishing a new strategy for asthma immunotherapy.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Chemokine CCL19; Chemokine CCL21; Chemokines, CC; Disease Models, Animal; Epitopes, T-Lymphocyte; Humans; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Inflammation Mediators; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Ovalbumin; Receptors, CCR7; Receptors, Chemokine; Receptors, Interleukin-2

Histamine plays an important role in immediate and late immune responses. The histamine type 1 (H1) receptor is expressed on several immune cell populations, but its role in a murine model of asthma remains unclear. The present study evaluated the role of histamine H1 receptors in airway allergic inflammation by comparing the development of bronchial asthma in histamine H1 receptor gene knockout (H1RKO) and wild-type mice.. H1RKO and wild-type mice were sensitized by intraperitoneal injection of ovalbumin (OVA) or saline, and then challenged with aerosolized OVA or saline. Ventilatory timing in response to inhaled methacholine was measured, and samples of blood, bronchoalveolar lavage, and lung tissues were taken 24 h after the last OVA challenge.. OVA-treatedwild-type mice showed significantly increased airway eosinophilic infiltration, and airway response to methacholine compared to OVA-treated H1RKO mice. The serum level of immunoglobulin E and levels of interleukin (IL)-4, IL-5, IL-13, and TGF-beta1 in bronchoalveolar lavage fluid were lower in OVA-treated H1RKO mice than in OVA-treated wild-type mice, but there was no significant difference in interferon-gamma expression. Overall, deletion of histamine H1 receptors reduced allergic responses in a murine model of bronchial asthma.. Histamine plays an important role via H1 receptors in the development of T helper type 2 responses to enhance airway inflammation.

Topics: Animals; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Histocytochemistry; Hypersensitivity; Immunoglobulin E; Lung; Male; Methacholine Chloride; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Histamine H1; Specific Pathogen-Free Organisms

Interleukin-1 (IL-1) is a proinflammatory cytokine and IL-1 receptor antagonist (IL-1ra) is a natural inhibitor that binds to IL-1 receptor type I without inducing signal transduction. It is suggested that IL-1 is required for allergen-specific T helper type 2 cell activation and the development of airway hyper-responsiveness (AHR), but the immunologic effect of exogenous IL-1ra in allergic asthma remains unclear. To examine the effect of IL-1ra on airway inflammation and immunoeffector cells in allergic asthma, recombinant adenovirus expressing human IL-1ra (Ad-hIL-1ra) was delivered intranasally into ovalbumin (OVA)-immunized mice. Single intranasal administration of Ad-hIL-1ra before airway antigen challenge in OVA-immunized mice significantly decreased the severity of AHR and reduced pulmonary infiltration of eosinophils and neutrophils. Suppression of IL-5 and eotaxin with concomitant enhancement of interferon gamma in bronchoalveolar lavage fluid was also noted in OVA-immunized mice by administration of Ad-hIL-1ra. In addition, histological studies showed that Ad-hIL-1ra was able to decrease OVA-induced peribronchial inflammation. Taken together, our results indicated that administration of Ad-hIL-1ra may have therapeutic potential for the immunomodulatory treatment of allergic asthma.

Topics: Adenoviridae; Administration, Inhalation; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokines, CC; Female; Genetic Engineering; Genetic Therapy; Genetic Vectors; Hypersensitivity; Interferon-gamma; Interleukin 1 Receptor Antagonist Protein; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Th2 Cells; Transduction, Genetic

It is well known that the immunoregulatory cytokine interleukin (IL)-10 inhibits the accessory function of human dendritic cells (DC) in vitro. Recently, we have shown that these IL-10 DC inhibit the production of T helper cell 1 (Th1) and T helper cell 2 (Th2) cytokines by T cells from atopic individuals in vitro. The current study was set out to analyze whether IL-10 DC also exert inhibitory effects in vivo in a murine model of allergy to ovalbumin adsorbed to the adjuvant aluminium hydroxide (OVA/alum).. OVA-pulsed or unpulsed bone marrow-derived DC, treated with IL-10 or left untreated during generation, were injected intravenously into BALB/c mice prior to and during OVA/alum sensitization, and sera and immune responses of mesenterial lymph node cells were analyzed. Additionally, bronchoalveolar lavage was performed after intranasal challenge with OVA.. Treatment of BALB/c mice with OVA-pulsed DC led to a significantly enhanced proliferation as well as Th2 (IL-4, IL-5), Th1 (interferon-gamma) and IL-10 cytokine production after restimulation of lymph node cells with OVA in vitro compared with OVA immunization alone. In contrast, using OVA-pulsed IL-10 DC for transfer, proliferation and cytokine production by lymph node cells were not enhanced. OVA-specific immunoglobulin G1 (IgG1) and IgG2a production were significantly increased after transfer of OVA-pulsed DC and OVA-pulsed IL-10 DC, respectively, whereas anti-OVA IgE production and airway eosinophilia remained unchanged.. Our data indicate that IL-10 treatment of DC decreases the Th1 and Th2 stimulatory capacity of DC but does not actually inhibit systemic (IgE) and local (airway inflammation) allergen-specific immune responses in a murine model of allergy.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Cell Proliferation; Cell Transplantation; Cytokines; Dendritic Cells; Disease Models, Animal; Female; Flow Cytometry; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Interleukin-10; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Allergic diseases are reported to be caused by a skew in the balance between T helper type 1 and 2 cells. Because some lactic acid bacteria have been shown to stimulate IL-12 (p70) production, which in turn shifts the balance between the T helper type 1 and 2 cell response from the latter to the former, they have the potential to either prevent or ameliorate disease conditions or both. They have therefore been extensively studied in the recent past for their probiotic activities. Nevertheless, much less information is available concerning the microbial factors that determine the strain-dependent ability to affect the production of cytokines. The objectives of our study were first to select potentially probiotic lactobacilli that strongly stimulate cytokine production in vitro, and then to determine whether the selected Lactobacillus strains could suppress antigen-specific IgE production in vivo by using allergic model animals. Finally, our investigation was extended to analyze which bacterial components were responsible for the observed biological activity. Twenty strains of heat-killed lactobacilli isolated from humans were screened for their stimulatory activity for the production of IL-12 (p70) by murine splenocytes in vitro. The results showed that some strains of Lactobacillus plantarum and Lactobacillus gasseri had a higher stimulatory activity for IL-12 (p70) production than the other lactobacilli tested; however, this effect was strain dependent rather than species dependent. Oral administration of the heat-killed strains that showed higher stimulatory activity for IL-12 (p70) production tended to reduce the serum antigen-specific IgE levels in ovalbumin-sensitized BALB/c mice compared with the controls. Among the lactobacilli tested, L. gasseri OLL2809 showed the highest activity in reducing the level of antigen-specific IgE. Furthermore, the stimulatory activity for IL-12 (p70) production was found to be reduced after treating the lactobacilli with N-acetyl-muramidase and to be positively correlated with the amount of peptidoglycan in the cells. The present data suggest that L. gasseri OLL2809 is a good candidate for potential probiotics in terms of either the prevention or amelioration of allergic diseases or both. In addition, the strain-dependent stimulatory activity for IL-12 (p70) production was found to be due, at least in part, to the amount of peptidoglycan present in the cells.

Topics: Animals; Cytokines; Endopeptidases; Freeze Drying; Hot Temperature; Hypersensitivity; Immunoglobulin E; Interleukin-12; Lactobacillus; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Peptidoglycan; Probiotics; Spleen; Teichoic Acids

We investigated the role of antioxidants in airway hyperresponsiveness to acetylcholine using young asthma model mice, which were sensitized and stimulated with ovalbumin.. The mice had been fed either a normal diet, an alpha-tocopherol-supplemented diet or a probucol-supplemented diet 14 days before the first sensitization. They were immunized with antigen at intervals of 12 days and, starting from 10 days after the second immunization, they were exposed to antigen 3 times every 4th day using an ultrasonic nebulizer. Twenty-four hours after the last antigen inhalation, airway responsiveness to acetylcholine was measured and bronchoalveolar lavage fluid (BALF) was collected. A blood and lung tissue study was also carried out.. Twenty-four hours after the last antigen challenge, both IL-4 and IL-5 in the BALF of alpha-tocopherol-supplemented mice were significantly decreased. The IL-5 level in probucol-supplemented mice was also decreased, but there was no difference in IL-4 levels. The serum IgE level was decreased in probucol-supplemented mice. Differential cell rates of the fluid revealed a significant decrease in eosinophils due to antioxidant supplementation. Airway hyperresponsiveness to acetylcholine was also repressed in antioxidant-supplemented mice. In histological sections of lung tissue, inflammatory cells and mucus secretion were markedly reduced in antioxidant-supplemented mice. We investigated the antioxidant effect on our model mice by examining 8-isoprostane in BALF and lung tissue, and acrolein in BALF; however, our experiment gave us no evidence of the antioxidant properties of either alpha-tocopherol or probucol contributing to the reduction of airway inflammation.. These findings indicate that alpha-tocopherol and probucol suppress allergic responses in asthma model mice, although these two drugs cause suppression in different ways that are unrelated to antioxidation.

Topics: Acrolein; Allergens; alpha-Tocopherol; Animals; Antioxidants; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Dietary Supplements; Dinoprost; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Probucol

This study was undertaken to investigate the involvement of cyclooxygenase-2 (COX-2) in allergic nasal inflammation in actively sensitized rats. An allergic rhinitis model was developed by the repeated topical application of antigen into the nasal cavities in the sensitized rats. The severity of allergic rhinitis was studied by measuring the nasal behavior, as well as electroencephalogram (EEG) activity by antigen challenge. The electrodes were implanted chronically into the bilateral olfactory bulb of the rats and the EEG was measured monopolarly with an electroencephalograph (EEG, Nohon Kohden, Japan). The intranasal application of antigen caused the increase of nasal allergic signs as well as an EEG spike in a dose-dependent fashion, and at a dose of 50 microg/site, it showed a significant effect. The responses induced by the antigen were evaluated with certain drugs, etodolac (a selective COX-2 inhibitor), indomethacin (a non-selective COX inhibitor), ramatroban (a thromboxane A2 receptor antagonist) and zafirlukast (a cys-leukotriene receptor antagonist). Etodolac showed the inhibition of nasal behavior and EEG spike in a dose-related fashion, and at doses of 3 and 10 mg/kg, it showed a significant effect. Moreover, ramatroban also caused the dose-related inhibition of nasal behavior and EEG spike induced by antigen. On the other hand, both indomethacin and zafirlukast had no effects on the responses induced by antigen, even at a higher dose. Therefore, it can be concluded that cyclooxygenase-2 actively participates in the allergic nasal inflammation in actively sensitized rats.

Topics: Animals; Carbazoles; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Disease Models, Animal; Electroencephalography; Etodolac; Hypersensitivity; Indoles; Indomethacin; Leukotriene Antagonists; Male; Olfactory Bulb; Ovalbumin; Phenylcarbamates; Rats; Rats, Wistar; Receptors, Thromboxane A2, Prostaglandin H2; Rhinitis; Sneezing; Sulfonamides; Tosyl Compounds

This study was undertaken to investigate the role of the aldose reductase in the refractoriness of diabetic rats to allergic inflammation. Wistar rats were actively sensitized with a mixture of Al(OH)3 plus ovalbumin and intrapleurally challenged with ovalbumin, 14 days later. Diabetes was induced by intravenous injection of alloxan into fasted rats, 7 days before sensitization, and the aldose reductase inhibitor zopolrestat was administered after 3 days of diabetes induction, once a day during 18 consecutive days. The treatment with zopolrestat restored antigen-induced protein extravazation and mast cell degranulation in the pleural cavity of diabetic sensitized rats. Zopolrestat also significantly reversed the suppression in the increase of total and specific levels of serum immunoglobulin E (IgE) noted in sensitized animals under conditions of diabetes. In addition, we noted that the drop in the pleural mast cell numbers as well as the increase in serum corticosterone levels in diabetic rats were inhibited by the drug. Our findings show that zopolrestat restored the hyporesponsiveness of diabetic rats to antigen provocation, in parallel with impairment of alloxan-induced mast cell depletion and hypercorticolism, indicating that polyol pathway activity seems to play an important role in these phenomena.

Topics: Aldehyde Reductase; Alloxan; Aluminum Hydroxide; Animals; Benzothiazoles; Cell Count; Cell Degranulation; Corticosterone; Diabetes Mellitus, Experimental; Hypersensitivity; Hypoglycemic Agents; Immunoglobulin G; Insulin; Male; Mast Cells; Ovalbumin; Phthalazines; Pleural Cavity; Proteins; Rats; Rats, Wistar

Pharmacological inhibition or genetic disruption of cyclooxygenase (COX)-1 or COX-2 exacerbates the inflammatory and functional responses of the lung to environmentally relevant stimuli. To further examine the contribution of COX-derived eicosanoids to basal lung function and to allergic lung inflammation, transgenic (Tr) mice were generated in which overexpression of human COX-1 was targeted to airway epithelium. Although no differences in basal respiratory or lung mechanical parameters were observed, COX-1 Tr mice had increased bronchoalveolar lavage fluid PGE(2) content compared with wild-type littermates (23.0 +/- 3.6 vs 8.4 +/- 1.4 pg/ml; p < 0.05) and exhibited decreased airway responsiveness to inhaled methacholine. In an OVA-induced allergic airway inflammation model, comparable up-regulation of COX-2 protein was observed in the lungs of allergic wild-type and COX-1 Tr mice. Furthermore, no genotype differences were observed in allergic mice in total cell number, eosinophil content (70 vs 76% of total cells, respectively), and inflammatory cytokine content of bronchoalveolar lavage fluid, or in airway responsiveness to inhaled methacholine (p > 0.05). To eliminate the presumed confounding effects of COX-2 up-regulation, COX-1 Tr mice were bred into a COX-2 null background. In these mice, the presence of the COX-1 transgene did not alter allergen-induced inflammation but significantly attenuated allergen-induced airway hyperresponsiveness, coincident with reduced airway leukotriene levels. Collectively, these data indicate that COX-1 overexpression attenuates airway responsiveness under basal conditions but does not influence allergic airway inflammation.

Topics: Animals; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cyclooxygenase 1; Cyclooxygenase 2; Dinoprostone; Humans; Hypersensitivity; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Transgenic; Ovalbumin; Plethysmography, Whole Body; Respiratory Function Tests; Reverse Transcriptase Polymerase Chain Reaction; Transgenes

Infections with pinworms are common in rodent animal facilities. In this study, we show the consequence of an outbreak in a transgenic barrier facility of infection by Syphacia obvelata, a murine pinworm gastrointestinal nematode. Immune responses were defined in experimental infection studies with BALB/c mice. Infection with S. obvelata induced a transient Th2-type immune response with elevated interleukin 4 (IL-4), IL-5, and IL-13 cytokine production and parasite-specific immunoglobulin G1 (IgG1). In contrast, BALB/c mice deficient in IL-13, IL-4/13, or the IL-4 receptor alpha chain showed chronic disease, with a >100-fold higher parasite burden, increased gamma interferon production, parasite-specific IgG2b, and a default Th2 response. Interestingly, infected IL-4-/- BALB/c mice showed only slightly elevated parasite burdens compared to the control mice, suggesting that IL-13 plays the dominant role in the control of S. obvelata. The influence that pinworm infection has on the allergic response to a dietary antigen was found to be important. Helminth-infected mice immunized against ovalbumin (Ova) elicited more severe anaphylactic shock with reduced Ova-specific IL-4 and IL-5 than did noninfected controls, demonstrating that S. obvelata infection is able to influence nonrelated laboratory experiments. The latter outcome highlights the importance of maintaining mice for use as experimental models under pinworm-free conditions.

Topics: Animals; Chemokines; Enterobiasis; Enterobius; Hypersensitivity; Immunoglobulin G; Interleukins; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Ovalbumin; Receptors, Cell Surface; Rodent Diseases; Th2 Cells

In mice, allergic rhinitis augments the infectious and inflammatory response to Streptococcus pneumoniae-induced sinusitis.. To investigate the effects of cysteinyl leukotriene antagonism on the severity of bacterial infection.. We performed 3 parallel, placebo-controlled experiments. In the first, mice were ovalbumin sensitized and ovalbumin challenged to show the effects of montelukast on the allergic inflammation; in the second, we evaluated the effect of montelukast on S. pneumoniae infection; in the third, we used mice that were both allergic and infected. Montelukast was given starting 2 days after sensitization until the day before euthanasia. One day after drug treatment began, the mice were inoculated intranasally with S. pneumoniae in the infected groups. Nasal hypersensitivity was measured with histamine challenges before the first sensitization and on the day before euthanasia. On the fifth day after infection, mice were euthanized, nasal lavage was performed, bacteria were cultured, and inflammatory cells in the sinuses were quantified.. Mice that were infected only tended toward having increased bacterial counts from nasal lavage in the montelukast-treated group. The mice that were allergic and infected experienced significantly higher bacterial counts (P < .05). All 3 montelukast treatment groups had significantly decreased eosinophil counts as well as T-lymphocyte counts.. Montelukast reduces the manifestations of allergic rhinitis in mice. Surprisingly, montelukast led to an increase in bacterial growth in infected mice. This suggests an effect of the cysteinyl leukotrienes on the innate response to bacterial infection.

Topics: Acetates; Animals; Cyclopropanes; Female; Flow Cytometry; Hypersensitivity; Leukotriene Antagonists; Male; Mice; Mice, Inbred BALB C; Nasal Lavage Fluid; Ovalbumin; Pneumococcal Infections; Quinolines; Sinusitis; Streptococcus pneumoniae; Sulfides

To study the importance of ongoing allergen exposure and TH1/TH2 genetic background in augmented bacterial and inflammatory responses in allergic and infected mice.. BALB/c and C57BL/6 mice were made allergic to ovalbumin. After 1 day of intranasal allergen exposure, they were inoculated intranasally with Streptococcus pneumoniae. The numbers of bacteria and inflammatory cells in the sinuses were determined, and nasal responsiveness to histamine was assessed.. Infected BALB/c and C57BL/6 mice that received ongoing ovalbumin challenge following intraperitoneal sensitization showed significantly greater bacterial load and phagocyte level compared with the infected-only mice. Differences were diminished after the allergen challenge was stopped. Allergic and infected C57BL/6 mice showed fewer bacteria and phagocytes compared with the allergic and infected BALB/c mice. Surprisingly, in contrast to the nonallergenic C57BL/6 mice, the infected BALB/c mice showed a larger number of bacteria 28 days after infection.. Ongoing allergic reaction augments bacterial load in both BALB/c and C57BL/6 mice and induces nasal hyperreactivity to histamine. Allergic and infected C57BL/6 mice show less allergic inflammation and bacterial load compared with allergic and infected BALB/c mice. Stopping allergen exposure reduces the response. Infected BALB/c mice, which favor a TH2 response, were less able to clear infection than C57BL/6 mice, which favor a TH1 response. Inflammation and bacterial load are affected by genetic background of mice and ongoing allergen stimulation.

Topics: Allergens; Animals; Colony Count, Microbial; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Nasal Cavity; Nasal Provocation Tests; Ovalbumin; Paranasal Sinuses; Pneumococcal Infections; Sinusitis; Streptococcus pneumoniae; Th1 Cells; Th2 Cells

Increased levels of macrophage migration inhibitory factor (MIF) in serum, sputum, and bronchioalveolar lavage fluid (BALF) from asthmatic patients and time/dose-dependent expression of MIF in eosinophils in response to phorbol myristate acetate suggest the participation of MIF in airway inflammation. In this study, we examined inflammation in OVA-sensitized mouse lungs in wild-type and MIF-deficient mice (MIF(-/-)). We report increased MIF in the lung and BALF of sensitized wild-type mice. MIF(-/-) mice demonstrated significant reductions in serum IgE and alveolar inflammatory cell recruitment. Reduced Th1/Th2 cytokines and chemokines also were detected in serum or BALF from MIF(-/-) mice. Importantly, alveolar macrophages and mast cells, but not dendritic cells or splenocytes, from MIF(-/-) mice demonstrated impaired CD4+ T cell activation, and the reconstitution of wild-type mast cells in MIF(-/-) mice restored the phenotype of OVA-induced airway inflammation, revealing a novel and essential role of mast cell-derived MIF in experimentally induced airway allergic diseases.

Topics: Animals; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Chemokines; Cytokines; Dendritic Cells; Hypersensitivity; Immunoglobulin E; Lung; Lymphocyte Activation; Macrophage Migration-Inhibitory Factors; Macrophages, Alveolar; Mast Cells; Mice; Mice, Mutant Strains; Ovalbumin; Pneumonia; Th1 Cells; Th2 Cells

Heme oxygenase-1 (HO-1) has anti-inflammatory effects in asthma. CD4+CD25(high) regulatory T cells (Treg) are a potent immunoregulator that suppresses the immune response. We studied the effects of HO-1-mediated CD4+CD25(high) Treg on suppression of allergic airway inflammation by comparing mice treated with hemin, OVA, Sn-protoporphyrin (SnPP), and hemin plus SnPP. Airway responsiveness, airway eosinophil infiltration, the level of OVA-specific IgE, and the numbers of cells in general and eosinophils in particular in bronchial alveolar lavage fluid were lower in the hemin group than in the OVA, SnPP, and hemin plus SnPP groups. The expressions of HO-1 mRNA and protein in the lung were increased by repeated administrations of hemin and SnPP. However, the activity of HO-1 was highest in hemin mice. The percentage and suppressive function of CD4+CD25(high) Treg and the expression of Foxp3 mRNA were obviously enhanced after treatment with hemin. This increase was diminished by the administration of SnPP. The concentration of serum IL-10 was higher in the hemin group than in the other groups, whereas the level of serum TGF-beta did not significantly differ across groups. Furthermore, the ratio of IFN-gamma/IL-4 mRNA in the lung was higher in hemin-treated mice than in OVA and SnPP mice. The suppressive capacity of CD4+CD25(high) Treg was not enhanced in the IL-10-deficient mice treated with hemin. In conclusion, our experiments in the animal model demonstrated that HO-1 has anti-inflammatory effects, probably via enhancement of the secretion of IL-10 and promotion of the percentage of CD4+CD25(high) Treg.

Topics: Animals; Asthma; CD4 Antigens; Disease Models, Animal; Eosinophils; Female; Forkhead Transcription Factors; Heme Oxygenase-1; Hemin; Hypersensitivity; Immunoglobulin E; Inflammation; Interferon-gamma; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Interleukin-4; Lung; Metalloporphyrins; Mice; Mice, Inbred BALB C; Ovalbumin; Protoporphyrins; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

In the present study, ovalbumin-sensitized/challenged rats were characterized by an nerve growth factor (NGF) increase in both serum and bronchial alveolar lavage fluid (BALF), but not in the lung. Exogenous administration of NGF or NGF-neutralizing antibodies did not modify immunoglobulin (IgE) and eosinophil parameters. In control rats, NGF administration did not induce increase of IgE or eosinophils in both BALF and lung. The present findings suggest that at least NGF does not act as a proper proinflammatory factor but most probably as a neuroimmune modulator molecule of the allergic state.

Topics: Animals; Antibodies; Bronchoalveolar Lavage Fluid; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Mast Cells; Nerve Growth Factor; Neuroimmunomodulation; Ovalbumin; Pneumonia; Rats; Rats, Sprague-Dawley; Tumor Necrosis Factor-alpha

Immunological oral tolerance is being studied with great interest due to its therapeutic potential in allergy and autoimmunity processes, although the cellular and molecular mechanisms linking these different phenomena remain elusive. In the present study, two mouse lines with extreme phenotypes for susceptibility [TS Line] or resistance [TR Line] to oral tolerance and their [TS x TR]F2 segregants were used in order to evaluate the impact of these traits on the atopic potential of the individuals.. Demonstrate whether the tr and ts genes, cumulated during 18 generations of bidirectional genetic selection, influence expression of two important immunobiological traits (IgE and mast cell) critical to allergic response.. Mice with extreme phenotypes for oral tolerance to ovalbumin (OVA), produced by assortative mating (TS and TR Line), and their (TS x TR)F2 segregating were used. Serum IgE levels assayed by ELISA, and mastocytes counted with toluidine blue staining were evaluated in naïve mice. Anaphylaxis was induced by intravenous injection of OVA, intestinal inflammation by oral administration of OVA 7 days after immunization, and pulmonary inflammation by intranasal and nebulization OVA challenges. Specific IgE was dosed by passive cutaneous anaphylaxis.. The naïve TS mice have a 20-fold lower serum IgE level and two- to threefold diminished mast cell numbers in mucosal sites, when compared with TR-mice, which were highly susceptible to allergic inflammation and anaphylactic shock. The associations of oral tolerance, serum IgE levels and mast cell numbers in naïve animals were confirmed analysing the simultaneous presence of these traits in individuals of a [TS x TR]F2 -segregating population.. The results suggest that the complex of genes controlling TS and TR phenotypes play a main role in the regulation of the atopic potential of the individual. The studies of these traits in interline F2 segregants demonstrated a co-segregation of TS and TR phenotypes with IgE responsiveness and mast cell numbers. Thus, the opposite capacity of the genetically modified mice may be involved in co-adaptative mechanisms reflecting a dynamic relation between gene frequencies in a natural population. These correlations give circumstantial evidence to support clinical applications of oral tolerance in allergic and autoimmune diseases.

Topics: Administration, Oral; Animals; Animals, Outbred Strains; Breeding; Bronchoalveolar Lavage Fluid; Cell Count; Genetic Predisposition to Disease; Genotype; Hypersensitivity; Immunity, Innate; Immunoglobulin E; Intestinal Mucosa; Mast Cells; Mice; Mucous Membrane; Ovalbumin; Passive Cutaneous Anaphylaxis; Phenotype; Respiratory Mucosa

The objectives of this study were to define the pharmacokinetics of recombinant human interleukin-1 receptor antagonist (rhIL-1ra) and its effects on allergic asthma, cell adhesion molecules, and upper respiratory tract following non-parenteral administration in animals. Pharmacokinetics and immunomodulating effects of rhIL-1ra were investigated in Sprague-Dawley rats and asthmatic guinea pigs, respectively. Effects on the upper respiratory tract following the applications of rhIL-1ra were investigated on the ex vivo nasal mucosa of Sprague-Dawley rats and in situ in the upper palate of Chinese toads. Absolute bioavailabilities after intratracheal and intranasal administrations of rhIL-1ra were 94.3% and 24.8%, respectively. After administration of rhIL-1ra solution as ultrasonic spraying, the asthmatic symptom in guinea pigs was obviously attenuated. The plasma soluble intercellular cell adhesion molecule (sICAM-1) and P-selectin levels in asthmatic guinea pigs were each dose-dependently reduced with the increase of rhIL-1ra dose. The rhIL-1ra solution after administration via the airway seemed to have no impact on the integrity of nasal mucosa and mucocilia clearance in the upper respiratory tract. The present study provides evidence that rhIL-1ra effectively suppresses allergen-induced asthmatic symptoms through spraying, which corresponds to nasal and pulmonary absorption or both, and the efficacy is associated with downregulation of sICAM-1 and P-selectin expressions.

Topics: Administration, Intranasal; Animals; Anti-Asthmatic Agents; Area Under Curve; Asthma; Biological Availability; Bufonidae; Cell Adhesion Molecules; Guinea Pigs; Humans; Hypersensitivity; Injections, Intravenous; Injections, Subcutaneous; Intercellular Adhesion Molecule-1; Interleukin 1 Receptor Antagonist Protein; Male; Mucociliary Clearance; Nasal Mucosa; Ovalbumin; P-Selectin; Rats; Rats, Sprague-Dawley; Recombinant Proteins; Respiratory System

Asthma is a complex inflammatory disorder involving obstruction, constriction, oedema, remodelling and hyperresponsiveness of the airways. These effects are induced by a raft of mediators, many of which exert their actions by stimulating specific G-protein-coupled receptors linked to a signal transduction pathway involving the monomeric GTPase; rho, and a downstream effector; rho kinase. The aim of this study was to determine whether administration of a selective inhibitor of rho kinase, Y-27632, attenuates airway inflammation, bronchoconstriction and hyperresponsiveness in a murine model of acute allergic inflammation. Intranasal administration of Y-27632 caused a dose-dependent inhibition in the number of eosinophils recovered from bronchoalveolar lavage fluid of ovalbumin-sensitised and challenged (allergic) mice. These inhibitory effects of intranasal Y-27632 on pulmonary eosinophilia were accompanied by a significant inhibition of the development of airways hyperresponsiveness in allergic mice. In additional studies, intranasal Y-27632 inhibited methacholine-induced increases in airways resistance in a time-dependent manner. In conclusion, these findings indicate that activation of rho kinase contributes to bronchoconstriction and eosinophil trafficking in murine models of acute allergic airway inflammation and to the development of airway hyperresponsiveness.

Topics: Administration, Intranasal; Aluminum Hydroxide; Amides; Animals; Australia; Bronchial Hyperreactivity; Bronchoalveolar Lavage; Bronchoconstriction; Disease Models, Animal; Female; Hypersensitivity; Injections, Intraperitoneal; Intracellular Signaling Peptides and Proteins; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Protein Serine-Threonine Kinases; Pulmonary Eosinophilia; Pyridines; rho-Associated Kinases

Nonselective cyclooxygenase (COX) inhibition during the development of allergic disease in a murine model causes an increase in type 2 cytokines and lung eosinophilia; however, the mechanisms responsible for this augmented allergen-induced inflammation have not been examined. Ab depletion of CD4 and CD8 cells revealed that the heightened allergic inflammation caused by COX inhibition was CD4, but not CD8, dependent. Allergen sensitization and airway challenge alone led to undetectable levels of IL-5 and IL-13 in the lungs of IL-4, IL-4Ralpha, and STAT6 knockout (KO) mice, but COX inhibition during the development of allergic inflammation resulted in wild-type levels of IL-5 and IL-13 and heightened airway eosinophilia in each of the three KO mice. These results indicate that the effect of COX inhibition was independent of signaling through IL-4, IL-4Ralpha, and STAT6. However, whereas COX inhibition increased IgE levels in allergic wild-type mice, IgE levels were undetectable in IL-4, IL-4Ralpha, and STAT6 KO mice, suggesting that IL-13 alone is not a switch factor for IgE synthesis in this model. These results illustrate the central role played by products derived from the COX pathway in the regulation of allergic immune responses.

Topics: Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cyclooxygenase Inhibitors; Cytokines; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Indomethacin; Inflammation; Interleukin-13; Interleukin-5; Lung Diseases; Mice; Mice, Knockout; Ovalbumin; Prostaglandin-Endoperoxide Synthases; Receptors, Interleukin-4; STAT6 Transcription Factor; Trans-Activators

The prevalence of asthma and allergic disease has increased in many countries and there has been speculation that immunization promotes allergic sensitization. Bordetella pertussis infection exacerbates allergic asthmatic responses. We investigated whether whole-cell pertussis vaccine (Pw) enhanced or prevented B. pertussis induced exacerbation of allergic asthma. Groups of mice were immunized with Pw, infected with B. pertussis and/or sensitized to ovalbumin. Immunological, pathological and physiological changes were measured to assess the impact of Pw immunization on immune deviation and airway function. Pw immunization modulated ovalbumin-specific serum IgE production, and reduced local and systemic IL-13 and other cytokine responses to sensitizing allergen. Histopathological examination revealed Pw immunization reduced the severity of airway pathology and decreased bronchial hyperreactivity to methacholine exposure. Pw does not enhance airway IL-13 and consequently does not enhance but protects against the exacerbation of allergic responses. We find no evidence of Pw contributing to allergic asthma, but rather provide evidence of a mechanism whereby whole-cell pertussis vaccination has a protective role.

Topics: Animals; Asthma; Bordetella pertussis; Hypersensitivity; Immunoglobulin E; Interleukin-10; Interleukin-13; Mice; Mice, Inbred BALB C; Ovalbumin; Pertussis Vaccine

MyD88 is a common Toll-like receptor (TLR) adaptor molecule found to be essential for induction of adaptive Th1 immunity. Conversely, innate control of adaptive Th2 immunity has been shown to occur in a MyD88-independent manner. In this study, we show that MyD88 is an essential innate component in the induction of TLR4-dependent Th2 responses to intranasal antigen; thus we demonstrate what we believe to be a novel role for MyD88 in pulmonary Th2 immunity. Induction of the MyD88-independent type I IFN response to LPS is defective in the pulmonary environment. Moreover, in the absence of MyD88, LPS-induced upregulation of costimulatory molecule expression on pulmonary DCs is defective, in contrast to what has been observed with bone marrow-derived DCs (BMDCs). Reconstitution of Th2 responses occurs upon adoptive pulmonary transfer of activated BMDCs to MyD88-deficient recipients. Furthermore, the dependence of Th2 responses on MyD88 is governed by the initial route of antigen exposure; this demonstrates what we believe are novel site-specific innate mechanisms for control of adaptive Th2 immunity.

Topics: Adaptor Proteins, Signal Transducing; Administration, Intranasal; Animals; Antigens, Differentiation; Bone Marrow Cells; Dendritic Cells; Gene Expression Regulation; Hypersensitivity; Immunity, Innate; Immunotherapy, Adoptive; Lipopolysaccharides; Lung; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Knockout; Myeloid Differentiation Factor 88; Ovalbumin; Receptors, Cell Surface; Receptors, Immunologic; Signal Transduction; Th2 Cells; Toll-Like Receptor 4

The cytokine IL-6 acts via a specific receptor complex that consists of the membrane-bound IL-6 receptor (mIL-6R) or the soluble IL-6 receptor (sIL-6R) and glycoprotein 130 (gp130). In this study, we investigated the role of IL-6R components in asthma. We observed increased levels of sIL-6R in the airways of patients with allergic asthma as compared to those in controls. In addition, local blockade of the sIL-6R in a murine model of late-phase asthma after OVA sensitization by gp130-fraction constant led to suppression of Th2 cells in the lung. By contrast, blockade of mIL-6R induced local expansion of Foxp3-positive CD4+CD25+ Tregs with increased immunosuppressive capacities. CD4+CD25+ but not CD4+CD25- lung T cells selectively expressed the IL-6R alpha chain and showed IL-6-dependent STAT-3 phosphorylation. Finally, in an in vivo transfer model of asthma in immunodeficient Rag1 mice, CD4+CD25+ T cells isolated from anti-IL-6R antibody-treated mice exhibited marked immunosuppressive and antiinflammatory functions. IL-6 signaling therefore controls the balance between effector cells and Tregs in the lung by means of different receptor components. Furthermore, inhibition of IL-6 signaling emerges as a novel molecular approach for the treatment of allergic asthma.

Topics: Adult; Animals; Antibodies; Asthma; DNA-Binding Proteins; Female; Forkhead Transcription Factors; Homeodomain Proteins; Humans; Hypersensitivity; Inflammation; Lung; Male; Mice; Mice, Knockout; Ovalbumin; Receptors, Cytokine; Receptors, Interleukin-2; Receptors, Interleukin-6; Signal Transduction; STAT3 Transcription Factor; Th2 Cells; Trans-Activators

We recently found that dietary raffinose suppressed allergic airway eosinophilia in ovalbumin-sensitized Brown Norway rats. Using this model in the present study, we compared the efficacy of other oligosaccharides with that of raffinose. Brown Norway rats were immunized s.c. with ovalbumin on d 0 and exposed to aerosolized ovalbumin on d 20; broncho-alveolar lavage fluid was obtained on d 21. In Expt. 1, rats were fed a control diet or diets supplemented with different oligosaccharides (50 g/kg diet, raffinose, alpha-linked galactooligosaccharide, fructooligosaccharide, and xylooligosaccharide). The number of eosinophils in the fluid was significantly lower in rats fed raffinose and alpha-linked galactooligosaccharide diets than in those fed the control diet. Dietary fructooligosaccharide and xylooligosaccharide did not affect airway eosinophilia. In Expt. 2, i.p. administration of raffinose and alpha-linked galactooligosaccharide, but not fructooligosaccharide and xylooligosaccharide, suppressed airway eosinophilia in rats fed the control diet. In Expt. 3, suppression of airway eosinophilia by dietary alpha-linked galactooligosaccharide occurred in cecectomized rats administered neomycin. Reduced levels of interleukin (IL)-4 and IL-5 mRNA in lung tissue were associated with the suppression of airway eosinophilia. We propose that indigestible oligosaccharides differ in their suppressive effect on allergic airway eosinophilia in ovalbumin-sensitized Brown Norway rats and that the effect appears not to be mediated by intestinal microflora.

Topics: Animals; Bronchoalveolar Lavage Fluid; Dietary Carbohydrates; Eosinophilia; Galactose; Hypersensitivity; Male; Oligosaccharides; Ovalbumin; Raffinose; Rats; Rats, Inbred BN

It is well-recognized that Stat6 plays a critical role in Th2 cell differentiation and the induction of allergic inflammation. We have previously shown that Stat5a is also required for Th2 cell differentiation and allergic airway inflammation. However, it is the relative importance and redundancy of Stat6 and Stat5a in Th2 cell differentiation and allergic airway inflammation are unknown. In this study we addressed these issues by comparing Stat5a-deficient (Stat5a(-/-)) mice, Stat6(-/-) mice, and Stat5a- and Stat6 double-deficient (Stat5a(-/-) Stat6(-/-)) mice on the same genetic background. Th2 cell differentiation was severely decreased in Stat6(-/-)CD4+ T cells, but Stat6-independent Th2 cell differentiation was still significantly observed in Stat6(-/-)CD4+ T cells. However, even in the Th2-polarizing condition (IL-4 plus anti-IFN-gamma mAb), no Th2 cells developed in Stat5a(-/-)Stat6(-/-) CD4+ T cells. Moreover, Ag-induced eosinophil and lymphocyte recruitment in the airways was severely decreased in Stat5a(-/-)Stat6(-/-) mice compared with that in Stat6(-/-) mice. These results indicate that Stat5a plays an indispensable role in Stat6-independent Th2 cell differentiation and subsequent Th2 cell-mediated allergic airway inflammation.

Topics: Animals; Cell Differentiation; DNA-Binding Proteins; Eosinophils; Female; Hypersensitivity; Inflammation; Lymphocytes; Male; Mice; Mice, Knockout; Milk Proteins; Ovalbumin; STAT5 Transcription Factor; STAT6 Transcription Factor; Th2 Cells; Trans-Activators

We have previously reported that, in IL-5-stimulated bone-marrow cultures, dexamethasone upregulates eosinophil differentiation and protects developing eosinophils from apoptosis induced by a variety of agents. Recently developed procedures for the isolation of hemopoietic cells from allergic murine lungs have enabled us to evaluate how these cells respond to dexamethasone in IL-5-stimulated cultures, when compared with bone-marrow-derived cells isolated from the same donors, and whether differences in response patterns were linked to apoptosis. Ovalbumin challenge of sensitized mice increased significantly the numbers of mature leukocytes as well as hemopoietic cells recovered from digested lung fragments, relative to saline-challenged, sensitized controls. Both mature eosinophils and cells capable of differentiating into eosinophils in the presence of IL-5 were present in lungs from sensitized mice 24 h after airway challenge. Dexamethasone strongly inhibited eosinophil differentiation in IL-5-stimulated cultures of lung hemopoietic cells. By contrast, dexamethasone enhanced eosinophil differentiation in cultures of allergic bone-marrow cells, in identical conditions. Hemopoietic cells from lungs and bone-marrow were respectively susceptible and resistant to induction of apoptosis by dexamethasone. The dexamethasone-sensitive step was the response to IL-5 in culture, while accumulation of IL-5 responsive cells in allergen-challenged lungs was dexamethasone-resistant. Cells from lungs and bone-marrow, cultured for 3 days with IL-5 in the absence of dexamethasone, did not respond to a subsequent exposure to dexamethasone in the presence of IL-5. These findings confirm that IL-5-responsive hemopoietic cells found in challenged, sensitized murine lungs differ from those in bone-marrow, with respect to the cellular responses induced by dexamethasone, including apoptosis.

Topics: Animals; Apoptosis; Bone Marrow Cells; Dexamethasone; Eosinophils; Hematopoietic Stem Cells; Hypersensitivity; Immunization; In Vitro Techniques; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Proteins

The cumulative activity of benzo(a)pyrene (BP) as one of the polycyclic aromatic hydrocarbons which is present in diesel exhaust particles on systemic immune responses was studied in mice after intranasal instillation and in vitro exposure to allergens. The main purpose was to elucidate whether BP has an adjuvant effect in this responses or not. Male Balb/cA mice were immunized with mite allergen extract (MAE) either alone or in combination with BP. Controls were given BP, or buffer alone. The animals were immunized four times. One week after the last immunization, anti-MAE IgE antibody in serum and IL-4, IL-10, IL-12p70 and IFN-gamma secreted by spleen cells upon rechallenge with the same allergen, was measured by ELISA. An increased response to the antigen was observed in animals receiving MAE together with BP, compared with animals receiving MAE alone. In another attempt, bone marrow-derived dendritic cells (DCs) are obtained from naive male Balb/c mice and were exposed to the varying doses of ovalbumin (OA) and BP. There were four groups of study: control, BP, OA and OA plus BP. An elevated response was observed in animals receiving OA together with BP, compared with animals receiving OA alone. In conclusion, this work shows that BP has an cumulative activity for specific IgE and Th1/Th2 cytokines production after intranasal instillation and cytokine secretion by DCs after in vitro exposure, and this phenomenon is dose-dependent. Thus, it may be another evidence on immunomodulatory effect of dendritic cells.

Topics: Administration, Intranasal; Air Pollutants; Allergens; Animals; Antibody Formation; Benzo(a)pyrene; Carcinogens; Cytokines; Dose-Response Relationship, Drug; Hypersensitivity; Immunization; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Mites; Models, Animal; Ovalbumin; Vehicle Emissions

alpha-Galactosylceramide (alpha-GalCer) is a specific ligand of natural killer T cells (NKT cells) that regulates the immune responses such as tumor rejection and autoimmunity by producing interferon (IFN)-gamma and interleukin (IL)-4. However, it has not been determined whether alpha-GalCer-activated NKT cells modulate allergic inflammation. Because alpha-GalCer induces a large amount of IFN-gamma production by NKT cells, we hypothesized that an in vivo administration of alpha-GalCer could inhibit allergic airway inflammation in mice. Strikingly, a single intraperitoneal injection of alpha-GalCer almost completely abrogated an infiltrate with eosinophils in the lung tissue as well as in the bronchoalveolar lavage. This inhibition of allergic inflammation was associated with a significant decrease in the levels of IL-4, IL-5, and IL-13 in bronchoalveolar lavage fluid and in the number of goblet cells. In addition, this ligand significantly inhibited airway hyperresponsiveness to inhaled methacholine and raised the serum levels of ovalbumin-specific IgG2a with a decrease in those of ovalbumin-specific IgE. In IFN-gamma knockout mice, however, alpha-GalCer failed to exert such inhibitory effects in this asthma model. These results indicate that alpha-GalCer prevents allergic airway inflammation possibly through IFN-gamma production by ligand-activated NKT cells, suggesting the potential therapeutic application of alpha-GalCer in asthma.

Topics: Animals; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Cytotoxicity, Immunologic; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Galactosylceramides; Goblet Cells; Hyperplasia; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Interferon-gamma; Killer Cells, Natural; Ligands; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; T-Lymphocytes; Th2 Cells; Time Factors

Oral tolerance is an important approach in allergic diseases and murine model can provide useful information to improve its understanding and therapeutic measures. To address the influence of non-related allergen sensitization in immunized mice with the mite Blomia tropicalis (Bt) or ovalbumin (OVA) or with both Bt/OVA allergens. Furthermore, we sought to verify oral tolerance effect in the Bt/OVA co-immunization model. Mice sensitized with Bt and then exposed to OVA developed an enhanced IgE response to both allergens; contrariwise, this effect was not observed when OVA-sensitization was prior to Bt-sensitization. Co-injection of Bt and OVA led to a dominant IgE response towards OVA over Bt, which was not observed when co-immunization was performed with a 240-fold less amount of OVA. Induction of oral tolerance with OVA, prior to co-immunization, suppressed IgE response to both allergens, probably as a consequence of the increased levels of IFN-gamma found in these animals. The results evidenced that, depending on allergenic potential, new allergen exposure may exert an adjuvant effect to the first allergen used in the sensitization. The bystander suppression to non-related allergens through oral tolerance should be a useful mechanism to control sensitization to new allergens.

Topics: Administration, Oral; Allergens; Animals; Antibodies; Bystander Effect; Cells, Cultured; Clonal Anergy; Cytokines; Female; Hypersensitivity; Immunization; Mice; Mites; Models, Immunological; Ovalbumin; Rats; Spleen

The phosphatidylinositol-3 kinase (PI3K) family of signaling enzymes plays a crucial role in leukocyte recruitment and activation and hence, likely regulates the induction and propagation phases of inflammation. However, little data have emerged showing a role for these processes in the resolution phase in models of in vivo inflammation. Here, we have evaluated the role of PI3K for the migration and survival of eosinophils in a model of allergic pleurisy in mice. Eosinophil accumulation in PI3Kgamma-deficient mice was inhibited at 48 h, as compared with wild-type mice but not at earlier time-points (6 and 24 h). Experiments with adoptive transfer of bone marrow showed that PI3Kgamma in eosinophils but not in non-bone marrow-derived cells was required for their accumulation. Systemic treatment with PI3K inhibitors before antigen challenge prevented the recruitment of eosinophils. This was associated with decreased Akt phosphorylation, interleukin-5 production, and eosinophil release from the bone marrow. Treatment with PI3K inhibitors 24 h after antigen challenge markedly cleared the accumulated eosinophils, an effect associated with inhibition of Akt phosphorylation and an increased number of apoptotic events. Altogether, our data demonstrate an important role of PI3Kgamma for the maintenance of eosinophilic inflammation in vivo, whereas other isoforms of PI3K may be relevant for the recruitment process.

Topics: Androstadienes; Animals; Cell Movement; Cell Survival; Chromones; Class Ib Phosphatidylinositol 3-Kinase; Eosinophils; Hypersensitivity; Inflammation; Isoenzymes; Mice; Morpholines; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Pleurisy; Wortmannin

Immunoglobulin E (IgE) plays a central role in IgE-mediated immediate type hypersensitivity. Since production of IgE depends on Th2, efforts to block IgE production and control allergic reactions include tolerization of Th2 or deviating development of Th2. We hypothesized that cytotoxic T lymphocytes targeting natural IgE peptides/MHC I complexes can eliminate IgE-producing cells and inhibit centrally IgE production. CTL to self-IgE peptides were elicited in mice immunized with nonameric p109-117, p113-121, and p103-141 (CHepsilon2 domain), which encompass both peptides with an OVA helper peptide (OVAp restricted for H-2d/b) in liposomes and presented by dendritic cells (DC). CTL from BALB/c lysed IgE peptide-pulsed P815 target as well as IgE-producing 26.82 hybridomas (H-2d). Natural tolerance to self-IgE peptides was tested in IgE sufficient (IgE +/+) as well as IgE-deficient (IgE -/-) 129/SvEv mice (H-2b). Comparable magnitude of CTL responses was observed in both strains immunized with p109-117 or p103-141 concomitantly with CD4 T-cell costimulation. CTL from 129/SvEv lysed not only IgE peptide-pulsed EL-4 but also IgE-producing B4 hybridomas (H-2b). This observation strongly suggests a correspondence of epitope of immunogenic peptide to that of physiologically processed IgE peptides presented on IgE-producing cells. Moreover, CTL were generated in 129/SvEv, immunized with the recombinant antigenized antibody in liposomes encompassing p107-123, p109-117, and p113-121 expressed in CDR3 of VH62/human gamma1. Polyclonal IgE production was inhibited by coincubation with MHC I-restricted CTL in vitro. Furthermore, antigen-specific IgE responses were inhibited in mice, immunized with p109-117 and p103-141 while IgG responses were not suppressed. Since IgE peptide sequences of CHepsilon2 are ubiquitous to all murine IgE heavy chain, peptides made as such can serve as a universal IgE vaccine to prevent allergy for a myriad of allergens in rodents. This observation suggests that similar human IgE peptides should be identified and employed to downregulate human IgE production.

Topics: Animals; Antibodies; B-Lymphocytes; CD40 Ligand; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cell Transplantation; Complementarity Determining Regions; Cytotoxicity Tests, Immunologic; Dendritic Cells; Down-Regulation; Female; H-2 Antigens; Hemocyanins; Histocompatibility Antigens Class I; Hybridomas; Hypersensitivity; Immunization; Immunoglobulin E; Immunotherapy, Active; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Peptide Fragments; Protein Binding; Recombinant Proteins; Self Tolerance; T-Lymphocytes, Cytotoxic

Brown Norway (BN) and Fischer 344 (F344) rats were exposed to aerosol of 1% ovalbumin (OVA) solution for 30 min at 1 week after the second sensitization with 1 mg of OVA at 2-week intervals. Changes in the histology and expression of cytokines and chemokines in the lung were examined for up to 96 h after the exposure. The lung weight significantly increased in BN rats but not in F344 rats. Histologically, in the lung of BN rats, multiple foci of hemorrhage in the alveolar space with infiltration of eosinophils and macrophages in the surrounding alveolar septa were first observed. Thereafter, granulomatous lesions developed in the preexisting hemorrhagic foci, finally resulting in formation of multiple eosinophilic granulomas. On the other hand, in F344 rats, infiltration of eosinophils and macrophages was observed around the vessels and bronchi. Thereafter it progressed gradually, resulting in mild thickening of alveolar septa. The levels of Th1- (interferon-gamma and interleukin 2 (IL-2)) and Th2-related cytokines (IL-4 and IL-5) and chemokines (eotaxin and monocyte chemoattractant protein-1) mRNAs measured by reverse transcription-polymerase chain reaction method were elevated in the lung of both strains, and the levels were higher in BN rats than in F344 rats. These results suggest that BN rats are more sensitive to OVA-sensitization/inhalation than F344 rats and that the difference in the severity of lung lesions between BN and F344 rats may reflect the difference in the expression levels of cytokines and chemokines between these two strains.

Topics: Animals; Chemokines; Disease Models, Animal; Granuloma; Hemorrhage; Hypersensitivity; Inhalation Exposure; Lung; Lung Diseases; Male; Organ Size; Ovalbumin; Rats; Rats, Inbred BN; Rats, Inbred F344; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Species Specificity; Trachea

The effect of Bacillus Calmette-Guérin (BCG) treatment in allergic pulmonary reaction was studied in mice genetically selected accordingly to a High (H-IVA) or Low (L-IVA) antibody responsiveness. Mice were immunized with ovalbumin (OVA) or OVA plus BCG. Two days after nasal antigenic challenge, seric IgE and IgG1 anti-OVA, eosinophils in pulmonary tissue, inflammatory cells in bronchoalveolar lavage and the compliance and conductance of respiratory system were evaluated. H-IVA mice were found more susceptible than L-IVA, and BCG was able to inhibit simultaneously the production of IgE, the bronchopulmonary inflammation and bronchial hyperresponsiveness in these genetically selected mice.

Topics: Animals; BCG Vaccine; Bronchial Hyperreactivity; Enzyme-Linked Immunosorbent Assay; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Interferon-gamma; Leukocytes; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Inbred WF

Mast cells are thought to be the main cause of an immediate asthmatic response, but their contribution to the late-phase of asthma is unknown.. To prove the contribution of preactivated mast cells to the late phase of allergic asthma by advanced activation.. Mast cell function in the late-phase of asthma was studied. Rats (wild, +/+ and mast cell deficient, Ws/Ws) were challenged with OVA to investigate the relationship between the contraction of airways and the population of inflammatory cells in the trachea.. During the entire asthmatic period, the contraction of the airway after OVA challenge in +/+ rats was enhanced significantly compared to Ws/Ws rats, especially in the late phase. The bronchoalveolar lavage fluid histamine in +/+, but not Ws/Ws, rats increased 5.3-fold in 30 min and 3.4-fold in 8 h after challenge, significantly. The number of mucosal mast cells in the tracheal epithelial layer in +/+ rats increased significantly 2.2-fold over controls at 8 h after challenge, as demonstrated by in situ hybridization.. Mast cells may contribute to the late phase of asthmatic response by continuous mast cell activation and the mucosal mast cell number increased in the late phase of asthmatic response.

Topics: Animals; Asthma; Blotting, Northern; Bronchoalveolar Lavage Fluid; Eosinophils; Histamine; Hypersensitivity; In Situ Hybridization; Inflammation; Male; Mast Cells; Ovalbumin; Rats; RNA; Time Factors; Trachea

Airway remodeling is a characteristic feature of asthma that leads to chronic irreversible airway obstruction. Fibrosis in the basement membrane region is a hallmark of remodeling in asthma that is not found in other diseases. In the outlined studies, we investigated the relationship between relaxin and airway fibrosis in asthma using acute and chronic models of allergic airway disease. These studies confirm a critical role for relaxin, in the regulation of collagen deposition in the airway/lung in animal models of allergic airway disease.

Topics: Acute Disease; Animals; Disease Models, Animal; Hypersensitivity; Mice; Mice, Knockout; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Pulmonary Fibrosis; Relaxin

Asthma is associated with increased expression of a typical array of genes involved in immune and inflammatory responses, including those encoding the prototypic Th2 cytokines interleukin (IL) 4, IL-5, and IL-13. Most of these genes contain binding sites for activator protein-1 (AP-1) within their promoter and are therefore believed to depend on AP-1 for their expression, suggesting that this transcription factor could be of particular importance in asthma pathophysiology.. To clarify the role of AP-1 in the effector phase of pulmonary allergy.. Ovalbumin (OVA)-sensitized mice were intratracheally given decoy oligodeoxyribonucleotides (ODNs) specifically directed to AP-1 or scrambled control ODNs before challenge with aerosolized OVA. Twenty-four hours after the last OVA challenge, airway hyperresponsiveness was measured and allergic airway inflammation was evaluated quantitatively. AP-1 decoys were localized using flow cytometry and immunohistochemistry. AP-1 activity in the lung was assessed using electrophoretic mobility shift assay.. Intratracheally delivered AP-1 decoys efficiently targeted airway immune cells, thus precluding AP-1 activation on OVA challenge. Decoy-mediated local inhibition of AP-1 resulted in significant attenuation of all the pathophysiologic features of experimental asthma-namely, eosinophilic airway inflammation, airway hyperresponsiveness, mucous cell hyperplasia, production of allergen-specific immunoglobulins, and synthesis of IL-4, IL-5, and IL-13. Scrambled control ODNs had no detectable effects.. Our results reveal a key role for AP-1 in the effector phase of pulmonary allergy and indicate that specific AP-1 inhibition in the airways may have therapeutic value in the control of established asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Cell Survival; Eosinophils; Female; Hypersensitivity; Immune System; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; Trachea; Transcription Factor AP-1

Adjuvants in vaccines are immune stimulants that play an important role in the induction of effective and appropriate immune responses to vaccine component(s). Diphtheria-tetanus-pertussis (DPT) vaccine contains not only aluminum hydrate (alum) to enhance the immune response to the vaccine ingredients, but also, both for that purpose and as a principal ingredient, pertussis toxin (PT). However, both adjuvants strongly promote T helper (Th) 2 type immune responses. Th1 and Th2 type immune responses are counterbalanced in vivo, and a Th2-prone immune response is not effective against intracellular infections but promotes IgE production, which is related to allergic disease. In this study, we used the CpG motif contained in oligodeoxynucleotide (CpG-ODN), which has an adjuvant effect and also induces the Th1 response, as an adjuvant to this vaccine, and we investigated its adjuvanticity and its potential to modulate immune responses to DPT vaccine. Administration of DPT vaccine with CpG-ODN (DPT-alum/ODN) to mice significantly reduced the total IgE levels and increased the anti-PT specific IgG2a titer in serum, in comparison with ordinary DPT vaccine (DPT-alum). Moreover, we investigated the antibody response to orally administrated ovalbumin (OVA) after vaccine administration. In the DPT-alum/ODN-administered group, the OVA specific IgE production in serum greatly decreased in comparison with that in the DPT-alum-administered group. These data indicate that CpG-ODN was not useful only as an efficient vaccine adjuvant but also shifted the immune responses substantially toward Th1 and modulated the Th1/Th2 immune response in DPT vaccine. These data suggested new applications of CpG-ODN as adjuvants in DPT vaccine.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Antigens; CpG Islands; Diphtheria-Tetanus-Pertussis Vaccine; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunity, Cellular; Immunoglobulin G; Intubation, Gastrointestinal; Mice; Mice, Inbred BALB C; Oligonucleotides; Ovalbumin; Th1 Cells; Th2 Cells

Tachykinins such as substance P are localized in unmyelinated slow-conducting C fibers that can be activated by noxious stimuli and tissue inflammation. Substance P is seldom expressed in fast-conducting large-diameter (A-fiber) vagal sensory neurons. We have previously found that allergic inflammation causes a phenotypic change in tachykinergic innervation of the trachea such that the production of substance P is induced in large-diameter sensory neurons projecting mechanosensitive A fibers to the trachea.. To evaluate whether allergic inflammation also induces substance P synthesis in large-diameter sensory stretch-receptor neurons innervating guinea pig lungs, and to investigate potential mechanisms by which this may occur.. Sensitized guinea pigs were exposed to allergen (ovalbumin) aerosol. One day later, immunohistochemical analysis was performed on vagal sensory neurons that had been retrogradely labeled from the lungs.. Ovalbumin inhalation caused a significant increase in substance P expression in large-diameter neurofilament-positive nodose ganglion neurons that innervate the lungs (P < .05). This effect was decreased by ipsilateral vagotomy. Exposing isolated nodose ganglia to the sensitizing antigen, ovalbumin, also significantly increased substance P expression compared with control.. Allergic inflammation induces substance P synthesis in large-diameter (A-fiber) nodose ganglion neurons innervating guinea pig lungs. This could contribute to the hyperreflexia seen in allergic airway disease. The full expression of this phenotypic switch in vagus nodose ganglion neurons requires intact vagus nerve, but if allergen reached the systemic circulation in sufficient quantities, it could also affect substance P synthesis by local activation of vagal ganglionic mast cells.

Topics: Allergens; Animals; Hypersensitivity; Lung; Male; Mice; Nodose Ganglion; Ovalbumin; Substance P; Vagotomy

The purpose of this study was to investigate the impact of mainstream cigarette smoke exposure (MTS) on allergic sensitization and the development of allergic inflammatory processes. Using two different experimental murine models of allergic airways inflammation, we present evidence that MTS increased cytokine production by splenocytes in response to OVA and ragweed challenge. Paradoxically, MTS exposure resulted in an overall attenuation of the immune inflammatory response, including a dramatic reduction in the number of eosinophils and activated (CD69+) and Th2-associated (T1ST2+) CD4 T lymphocytes in the lung. Although MTS did not impact circulating levels of OVA-specific IgE and IgG1, we observed a striking reduction in OVA-specific IgG2a production and significantly diminished airway hyperresponsiveness. MTS, therefore, plays a disparate role in the development of allergic responses, inducing a heightened state of allergen-specific sensitization, but dampening local immune inflammatory processes in the lung.

Topics: Allergens; Ambrosia; Animals; Antibody Formation; Bronchial Hyperreactivity; Cytokines; Female; Flow Cytometry; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Nicotiana; Ovalbumin; Smoke; Tobacco Smoke Pollution

Nigella sativa Linn. (Ranunculaceae) is known to have beneficial effects on a wide range of diseases including asthma. However, the mechanism of action in asthma and other allergic diseases is not entirely clear. The present study was planned to evaluate the effects of Nigella sativa on cytokine production of splenic mononuclear cells in ova-sensitized mice. Nineteen two-month-old BALB/c mice were given 0.3 mL of Nigella sativa oil by oro-eosophageal cannula once a day for a month. The control group consisting of 10 mice took 0.3 mL of 0.9% saline solution by the same route for the same period. In the third week of the study, all mice were sensitized by means of intraperitoneal injections of 20 microg of ovalbumin (OVA-Grade VI, Sigma). Ova injections were repeated three times with 7-day intervals. After another week, all mice were sacrificed by means of cervical dislocation. Then the splenic mononuclear cells (MNCs) of mice were cultured with OVA or Concavalin A (Con-A). From the culture supernatants, IL-4, IL-10 and IFN-gamma were assessed by means of ELISA. The cytokine production of splenic MNCs of mice that were given Nigella sativa for 30 days was not significantly different than those who took saline solution instead. In conclusion, Nigella sativa oil seems not to have an immunomodulatory effect on Th1 and Th2 cell responsiveness to allergen stimulation.

Topics: Allergens; Animals; Concanavalin A; Hypersensitivity; Immunologic Factors; Mice; Mice, Inbred BALB C; Monocytes; Nigella; Ovalbumin; Plant Oils; Spleen; Th1 Cells; Th2 Cells; Thymidine

Recent studies have highlighted the influence of fetal/maternal interactions on the development of asthma. Because IFN-gamma reduces Th2-mediated allergic responses, we assessed its capacity to modulate asthma in the offspring when injected into mothers during pregnancy. IFN-gamma was injected in CD1 female mice on day 6.5 of gestation. Immediately after birth, male newborns were housed in cages with interchanged mothers: the offspring from IFN-gamma-treated mothers were breastfed by normal mothers (IFN/nor), and those from normal mothers were breastfed by IFN-gamma-treated (Nor/IFN) or normal mothers (Nor/nor). Immediately after weaning, the spleen cells from IFN/nor and Nor/IFN mice produced less IL-4 and more IFN-gamma than Nor/nor mice when stimulated with Con A. At the age of 6-7 wk, mice were immunized with OVA on days 0 and 7. From day 14 to 16, they were exposed to aerosolized OVA. The bronchoalveolar lavage fluid from Nor/nor mice showed eosinophilia, a large number of these cells being present in perivascular and peribronchial regions of lung tissues. IFN/nor or Nor/IFN mice showed greatly reduced eosinophil numbers in bronchoalveolar lavage fluid. In addition, lung sections from IFN/nor, but not Nor/IFN mice showed almost normal histology. In OVA-sensitized IFN/nor and Nor/IFN mice, the production of IFN-gamma, IL-4, and IL-5 by spleen cells was significantly reduced as compared with cells from the OVA-sensitized Nor/nor group. IgE and anaphylactic IgG1 were also reduced in plasma of IFN/nor mice. In conclusion, the presence of IFN-gamma during pregnancy confers to the fetus a protection against allergenic provocations in the adult life.

Topics: Animals; Animals, Newborn; Asthma; Eosinophilia; Female; Hypersensitivity; Immunization; Interferon-gamma; Interleukin-4; Interleukin-5; Male; Maternal-Fetal Exchange; Mice; Mice, Inbred Strains; Ovalbumin; Pregnancy

To observe the effect on infantile allergic cough with Minkeqing oral liquid (Minkeqing) and to study its cell molecular biologic mechanism.. The rat model was induced by inhalating ovalbumin; then the effects of Minkeqing on IL-6, IL-8, ET-1, TX-B2 in the blood and the bronchoalveolar lavage fluid (BALF) of the animal model were observed.. Minkeqing could reduce the levels of IL-6,IL-8,ET-1,Tx-B2 in the blood and BALF of the animal model.. Minkeqing has the significant function of inhibiting the release of inflammatory mediums.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cough; Drug Combinations; Drugs, Chinese Herbal; Endothelin-1; Hypersensitivity; Interleukin-6; Interleukin-8; Male; Ovalbumin; Plants, Medicinal; Random Allocation; Rats; Rats, Wistar; Thromboxane B2

Allergic asthma is a multifaceted syndrome consisting of eosinophil-rich airway inflammation, bronchospasm, and airway hyper-responsiveness (AHR). Using a mouse model of allergic asthma, we previously reported that invariant NKT (iNKT) cells increase the severity of this disease. Herein, we demonstrate that a single i.v. injection of alpha-galactosylceramide (alpha-GalCer), 1 h before the first airway allergen challenge of OVA-sensitized mice, abrogates elicitation of AHR, airway eosinophilia, IL-4 and IL-5 production in bronchoalveolar lavage fluid, and specific anti-OVA IgE antibodies. Further, alpha-GalCer administered intranasally also strongly inhibited the major symptoms of asthma in sensitized and challenged mice. Alpha-GalCer treatment induces iNKT cell accumulation in the lungs, and shifts their cytokine profile from pro-asthmatic IL-4 to a protective IFN-gamma production. The role of IFN-gamma from iNKT cells in protection was shown by adoptive transfer of sorted iNKT cells from OVA-sensitized and alpha-GalCer-treated mice which protected immunized recipients from manifesting asthma by an IFN-gamma-dependent pathway. Our findings demonstrate for the first time that alpha-GalCer administered locally inhibits asthma symptoms, even in predisposed asthmatic mice, through an iNKT cell- and IFN-gamma-dependent pathway.

Topics: Administration, Intranasal; Adoptive Transfer; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Flow Cytometry; Galactosylceramides; Hypersensitivity; Injections, Intravenous; Interferon-alpha; Killer Cells, Natural; Male; Mice; Ovalbumin

Allergic airway diseases are related to exposure to atmospheric pollutants, which have been suggested to be one factor in the increasing prevalence of asthma. Little is known about the effect of ozone and diesel exhaust particulates (DEP) on the development or aggravation of asthma. We have used a mouse asthma model to determine the effect of ozone and DEP on airway hyperresponsiveness and inflammation. Methacholine enhanced pause (P(enh)) was measured. Levels of IL-4 and IFN-gamma were quantified in bronchoalveolar lavage fluids by enzyme immunoassays. The OVA-sensitized-challenged and ozone and DEP exposure group had higher P(enh) than the OVA-sensitized-challenged group and the OVA-sensitized-challenged and DEP exposure group, and the OVA-sensitized-challenged and ozone exposure group. Levels of IFN-gamma were decreased in the OVA-sensitized-challenged and DEP exposure group and the OVA-sensitized-challenged and ozone and DEP exposure group compared to the OVA-sensitized-challenged and ozone exposure group. Levels of IL-4 were increased in the OVA-sensitized-challenged and ozone exposure group and the OVA-sensitized-challenged and DEP exposure group, and the OVA-sensitized-challenged and ozone and DEP exposure group compared to OVA-sensitized-challenged group. Co-exposure of ozone and DEP has additive effect on airway hyperresponsiveness by modulation of IL-4 and IFN-gamma suggesting that DEP amplify Th2 immune response.

Topics: Air Pollutants; Animals; Asthma; Disease Models, Animal; Drug Combinations; Drug Synergism; Female; Hypersensitivity; Interferon-gamma; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Ozone; Pneumonia; Respiratory Hypersensitivity; Vehicle Emissions

We studied the effect of rolipram, a phosphodiesterase (PDE) IV inhibitor, on allergic footpad swelling in mice. For this study, varying adjuvants including complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA) and Imject Alum (Alum) were used because the extent of antigen-specifically induced T helper type 1 (Th1) and Th2 responses had been shown to depend on adjuvants used. To induce allergic footpad swelling, we immunized mice with ovalbumin (OVA) emulsified in either CFA or IFA, dissolved in Alum or in phosphate-buffered saline (PBS) as a control (day 0), followed by subcutaneous injection of the antigen into footpads on day 21. Rolipram was given orally to the animals daily from days 0-20. Results showed that treatment with rolipram was followed by an increase in early swelling at 0.5 h and a decrease in late swelling at 6 and 24 h in the CFA group. In the IFA group, rolipram significantly enhanced swelling at, but not after, 30 min. In the Alum and the PBS groups, the PDE inhibitor failed to affect the OVA-specific footpad reaction at all times examined. Treatment of the CFA and IFA groups with rolipram significantly inhibited the production of the Th1 antibody anti-OVA immunoglobulin G2a (IgG2a), and the drug enhanced Th2 cell-dependent anti-OVA IgE production. In both groups, rolipram also enhanced the secretion of Th2 cytokines including interleukin-4 (IL-4) and IL-10. These findings suggest that rolipram may facilitate early allergic footpad swelling mediated by Th2 immune responses, while the late phase of swelling associated with Th1 responses may be attenuated by the PDE IV inhibitor.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Adjuvants, Immunologic; Animals; Cells, Cultured; Cyclic Nucleotide Phosphodiesterases, Type 4; Edema; Foot; Hypersensitivity; Male; Mice; Mice, Inbred DBA; Ovalbumin; Phosphodiesterase Inhibitors; Rolipram; Spleen; Th1 Cells; Th2 Cells

New pathophysiologic concepts are needed to explain the clinically observed connection between the allergic diathesis and otitis media. Although mast cells, unlike lymphocytes, are common in the normal middle ear mucosa, their potential role in innate immunity of the middle ear and in the expression of inflammatory responses in that space to bacterial challenge, as opposed to allergy, has received relatively little attention.. In the current study, we examine the contributions of mast cells to the pathogenesis of bacterially induced inflammation in the middle ear and thus to otitis media.. Wild-type mice, mast cell-deficient mice, and mast cell-deficient mice whose mast cell populations were restored by transplantation of bone marrow-derived mast cells were challenged by using models of bacterial and allergic middle ear inflammation.. Our results indicate that mast cells account for a substantial proportion of the innate immune response to bacteria in the middle ear.. This mechanism may link responses to allergy and infection in the middle ear mucosa, and thus the mast cell may be a critical control element in the pathogenesis of otitis media.

Topics: Animals; Bone Marrow Cells; Cells, Cultured; Drug Combinations; Ear, Middle; Female; Haemophilus Infections; Haemophilus influenzae; Hypersensitivity; Injections; Mast Cells; Mice; Mice, Congenic; Mice, Mutant Strains; Otitis Media; Ovalbumin

Bromelain, a clinically used pineapple extract and natural product, has reported anti-inflammatory and immunomodulatory activities. The purpose of this study was to determine the effect of bromelain treatment in an ovalbumin (OVA)-induced murine model of allergic airway disease (AAD).. To establish AAD, mice were sensitized with intraperitoneal (i.p.) OVA/alum and challenged with daily OVA aerosols. Mice were treated i.p. with either saline, 2 or 6 mg/kg bromelain, twice daily for four consecutive days. Bronchoalveolar lavage leukocytes and cytokines, lung histology, airway hyperresponsiveness, and lymphocyte populations via flow cytometry were compared between groups.. Bromelain treatment of AAD mice resulted in reduced total BAL leukocytes, eosinophils, CD4+ and CD8+ T lymphocytes, CD4+/CD8+ T cell ratio, and IL-13.. Bromelain attenuated development of AAD while altering CD4+ to CD8+ T lymphocyte populations. The reduction in AAD outcomes suggests that bromelain may have similar effects in the treatment of human asthma and hypersensitivity disorders.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Bromelains; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Flow Cytometry; Fluorescent Antibody Technique; Hyaluronan Receptors; Hypersensitivity; Mice; Mice, Inbred C57BL; Ovalbumin

Asthma is associated with airway remodeling. Evidence of platelet recruitment to the lungs of asthmatics after allergen exposure suggests platelets participate in various aspects of asthma; although their importance is unknown in the context of airway remodeling, their involvement in atherosclerosis is established. Studies from our laboratory have shown a requirement for platelets in pulmonary leukocyte recruitment in a murine model of allergic lung inflammation. Presently, the effects of platelet depletion and corticosteroid administration on airway remodeling and lung function were examined. Ovalbumin (OVA)-sensitized mice, exposed to aerosolized OVA for 8 weeks, demonstrated epithelial and smooth muscle thickening, and subepithelial reticular fiber deposition in the distal airways. The depletion of platelets via an immunologic (antiplatelet antisera) or nonimmunologic (busulfan) method, markedly reduced airway remodeling. In contrast, dexamethasone administration did not affect epithelial thickening or subepithelial fibrosis, despite significantly inhibiting leukocyte recruitment. Thus, pathways leading to certain aspects of airway remodeling may not depend on leukocyte recruitment, whereas platelet activation is obligatory. OVA-sensitized mice exhibited airway hyperresponsiveness (AHR) compared with sham-sensitized mice following chronic OVA exposure. Neither platelet depletion nor dexamethasone administration inhibited chronic AHR; thus, mechanisms other than inflammation and airway remodeling may be involved in the pathogenesis of chronic AHR.

Topics: Adrenal Cortex Hormones; Animals; Asthma; Blood Platelets; Bronchoalveolar Lavage Fluid; Dexamethasone; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Inflammation; Leukocytes; Mice; Mice, Inbred C57BL; Ovalbumin; Platelet Count; Pulmonary Circulation; Respiratory Function Tests; Respiratory Mucosa; Thrombocytopenia

One third of asthmatic women report a decreased expiratory peak flow during menses. Since asthma is characterized by lung inflammation and bronchopulmonary hyperresponsiveness, we investigated the role played by estradiol in allergic lung inflammation.. Cell migration to the lungs of allergic female rats subjected to oophorectomy (OVx) was compared to that in their sham-operated (sham) control counterparts. Seven days after OVx or sham operation, the rats were sensitized intraperitoneally with ovalbumin (OA, 1 mg/kg) suspended in aluminum hydroxide (day 0). At day 7, a subcutaneous booster of OA was performed and an aerosolized OA challenge was carried out at day 14. One day later (day 15), the rats were killed and cell counts were performed in bronchoalveolar lavages (BAL), in peripheral blood and in bone marrow lavages.. After the antigen challenge, OVx rats showed a significant decrease in cell migration to the lung as compared to sham-operated rats. Differential analyses of BAL revealed a reduced number of eosinophils, mononuclear cells and neutrophils. In contrast, in bone marrow as well as in the peripheral blood the numbers of eosinophils, mononuclear cells and neutrophils were increased relative to sham controls. Mast cell numbers were similar in both groups. The estradiol receptor antagonist tamoxifen decreased the allergic lung inflammation in intact rats down to levels similar to those found in untreated OVx rats. In contrast, 17beta-estradiol replacement in OVx rats reestablished the allergic lung inflammation, as observed by an elevated number of eosinophils, mononuclear cells and neutrophils recovered in BAL. Similarly, an elevated number of inflammatory cells were quantified in BAL from allergic OVx rats when corticosterone effects were blocked with metyrapone or RU-486.. Our results suggest that estradiol has proinflammatory actions on the allergic lung response, and these actions seem to be mediated, at least in part, by endogenous glucocorticoids.

Topics: Animals; Antibodies; Asthma; Bone Marrow Cells; Cell Movement; Corticosterone; Estradiol; Female; Hypersensitivity; Leukocytes; Mast Cells; Neuroimmunomodulation; Ovalbumin; Ovariectomy; Pneumonia; Rats; Rats, Wistar

Asthma is one of the most common diseases and is characterized by airway obstruction, airway inflammation, and increased airway responsiveness. Glucocorticoids are very effective in treatment, but their long-term use is associated with several side effects, so that new anti-inflammatory drugs are in development. Activated protein C (APC) is a serine protease with potent anti-inflammatory effects. This study evaluated the effect of inhaled APC on airway inflammation and hyperresponsiveness in a murine asthma model. Asthma was induced in BALB/c mice by exposure to chicken egg ovalbumin (OVA), and the effect of inhaled APC was assessed by administering prior to OVA exposure. Inhalation of APC significantly inhibited the expression of T helper 2 (Th2) cytokines, immunoglobulin E (IgE), eosinophilic inflammation, and hyperresponsiveness. APC also significantly suppressed the expression of Th2 cytokines and IgE from lymphocytes isolated from OVA-sensitized/challenged animals. In addition, binding of signal transducer and activator of transcription 6 (STAT6) and nuclear factor kappa B (NF-kappa B) oligonucleotides to lung nuclear proteins was significantly reduced in mice treated with inhaled APC. In brief, the exogenous supplementation of APC inhibits the immunologic and inflammatory responses induced by Th2 cytokines in a mouse model of asthma and may represent a novel anti-inflammatory treatment.

Topics: Animals; Antibodies; Antigens, CD; Asthma; Biomarkers; Blood Coagulation Factors; Bronchial Hyperreactivity; Cytokines; Dinoprostone; Disease Models, Animal; Dose-Response Relationship, Drug; Endothelial Protein C Receptor; Endothelins; Eosinophils; Female; Glycoproteins; HeLa Cells; Humans; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; NF-kappa B; Nitric Oxide; Ovalbumin; Protein C; Receptor, PAR-1; Receptors, Cell Surface; Specific Pathogen-Free Organisms; STAT6 Transcription Factor; Th2 Cells; Thrombin; Trans-Activators; U937 Cells

Mice with allergically inflamed airways are widely used as animal models of asthma, but their relevance for human asthma is not understood. We, therefore, examined the time course of changes in respiratory input impedance during induced bronchoconstriction in BALB/c mice sensitized and challenged with ovalbumin. Our results indicate that bronchoconstriction in mice is accompanied by complete closure of substantial regions of the lung and that closure increases markedly when the lungs are allergically inflamed. With the aid of an anatomically accurate computational model of the mouse lung, we show that the hyperresponsiveness of mice with allergically inflamed airways can be explained entirely by a thickening of the airway mucosa and an increased propensity of the airways to close, without the involvement of any increase in the degree of airway smooth muscle shortening. This has implications for the pathophysiology of asthma and suggests that at least some types of asthma may benefit from therapies aimed at manipulating surface tension at the air-liquid interface in the lungs.

Topics: Animals; Asthma; Computer Simulation; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Models, Biological; Muscle, Smooth; Ovalbumin; Respiratory Physiological Phenomena; Respiratory System

1. The contribution of endogenous endothelins to nociceptive responses elicited by ovalbumin (OVA) in the hind-paw of mice sensitised to this antigen (50 microg OVA+5 mg Al(OH)(3), s.c., 14 days beforehand) was investigated. 2. Sensitised mice exhibited greater nocifensive responsiveness to intraplantar (i.pl.) OVA (total licking time over first 30 min: 85.2+/-14.6 s at 0.3 microg; 152.6+/-35.6 s at 1 microg) than nonsensitised animals (29.3+/-7.4 s at 1 microg). Nocifensive responses of sensitised mice to 0.3 microg OVA were inhibited by morphine (3 mg kg(-1), s.c.) or local depletion of mast cells (four daily i.pl. injections of compound 48/80). 3. Pretreatment with i.v. bosentan (mixed ET(A)/ET(B) receptor antagonist; 52 micromol kg(-1)) or A-122722.5 (selective ET(A) receptor antagonist; 6 micromol kg(-1)) reduced OVA-induced licking from 124.8+/-20.6 s to 45.7+/-13.0 s and 64.2+/-12.1 s, respectively, whereas A-192621.1 (selective ET(B) receptor antagonist; 25 micromol kg(-1)) enhanced them to 259.2+/-39.6 s. 4. Local i.pl. pretreatment with BQ-123 or BQ-788 (selective ET(A) or ET(B) receptor antagonists, respectively, each at 3 nmol) reduced OVA-induced licking (from 106.2+/-15.2 to 57.0+/-9.4 s and from 118.6+/-10.5 to 76.8+/-14.7 s, respectively). Sarafotoxin S6c (selective ETB receptor agonist, 30 pmol, i.pl., 30 min after OVA) induced nocifensive responses in OVA-sensitised, but not in nonsensitised, animals. 5. Compound 48/80 (0.3 microg, i.pl.) induced nocifensive responses per se and potentiated those induced by i.pl. capsaicin (0.1 microg). Treatment with BQ-123 (3 nmol, i.pl.) reduced only the hyperalgesic effect of compound 48/80, whereas BQ-788 (3 nmol) was ineffective. 6. Thus, immune-mediated Type I hypersensitivity reactions elicit mast cell- and endothelin-dependent nociception in the mouse hind-paw, which are mediated locally by both ET(A) and ET(B) receptors. The nocifensive response to antigen is amenable to blockade by systemic treatment with dual ET(A)/ET(B) or selective ET(A) receptor antagonists, but is sharply potentiated by systemic selective ET(B) receptor antagonist treatment. The apparently distinct roles played by ET(B) receptors in this phenomenon at local and other sites remain to be characterised.

Topics: Animals; Antigens; Bosentan; Dose-Response Relationship, Drug; Endothelins; Hyperalgesia; Hypersensitivity; Indicators and Reagents; Male; Mice; Oligopeptides; Ovalbumin; p-Methoxy-N-methylphenethylamine; Pain; Peptides, Cyclic; Piperidines; Sulfonamides; Viper Venoms

Various CC chemokine receptors are expressed on effector cells in allergic inflammation and their distinct expression pattern may dictate, to a large extent, the migration of inflammatory cells to sites of airway inflammation. The lipopolysaccharide (LPS)-inducible CC chemokine receptor (L-CCR) is an orphan chemokine receptor that has previously been identified in the murine macrophage cell line RAW 264.7 and in murine brain glial cells. In this study we investigated the induction and localization of L-CCR mRNA expression in mouse lung after ovalbumin (OVA)-induced airway inflammation. Both RT-PCR experiments and in situ hybridization (ISH) experiments in whole lung sections revealed a rapid upregulation of L-CCR mRNA expression as early as 1 hr and 3 hr after OVA challenge. Expression was found predominantly in MAC3(+) macrophages and in bronchial epithelium, as shown by ISH and immunohistochemistry (IHC). We demonstrated that L-CCR mRNA expression is strongly upregulated in mouse lung after OVA challenge and is localized in macrophages and bronchial epithelium. Regarding the likely role of L-CCR as a chemokine receptor with the putative ligand monocyte chemotactic protein-1 (MCP-1, CCL2), this receptor may have an important function in the early phase of airway inflammation.

Topics: Animals; Hypersensitivity; Immunohistochemistry; In Situ Hybridization; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Receptors, CCR; Receptors, CCR2; Receptors, Chemokine; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Respiratory viral infections in early childhood may interact with the immune system and modify allergen sensitization and/or allergic manifestations. In mice, respiratory syncytial virus (RSV) infection during allergic provocation aggravates the allergic T helper (Th) 2 immune response, characterized by the production of IL-4, IL-5, and IL-13, and inflammatory infiltrates. However, it is unclear whether the RSV-enhanced respiratory allergic response is a result of non-specific virus-induced damage of the lung, or virus-specific immune responses.. In the present study we investigated whether RSV, pneumonia virus of mice (PVM) and influenza A virus similarly affect the allergic response.. BALB/c mice were sensitized and challenged with ovalbumin (OVA), and inoculated with virus during the challenge period. Pulmonary inflammation, lung cytokine mRNA responses, and IgE production in serum were assessed after the last OVA-challenge.. Like RSV, PVM enhanced the OVA-induced pulmonary IL-4, IL-5, and IL-13 mRNA expression, which was associated with enhanced perivascular inflammation. In addition, PVM increased the influx of eosinophils in lung tissue. In contrast, influenza virus decreased the Th2 cytokine mRNA expression in the lungs. However, like PVM, influenza virus enhanced the pulmonary eosinophilic infiltration in OVA-allergic mice.. The Paramyxoviruses RSV and PVM both are able to enhance the allergic Th2 cytokine response and perivascular inflammation in BALB/c mice, while the Orthomyxovirus influenza A is not.

Topics: Animals; Female; Hypersensitivity; Immunoglobulin E; Influenza A virus; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Murine pneumonia virus; Orthomyxoviridae Infections; Ovalbumin; Pneumovirus Infections; Pulmonary Eosinophilia; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human; RNA, Messenger; Virus Diseases

The effect of bradykinin (B(1) or B(2)) receptor antagonists was studied in allergic and immune-complex-induced lung inflammation.. Lungs of BALB/c mice were examined 24 h after induction of lung inflammation, either allergic (ovalbumin-sensitized submitted to two aerosol of antigen, one week apart) or immune-complex induced (intratracheal instillation of IgG antibodies followed by intravenous antigen). The bradykinin B(2) receptor antagonist, HOE-140 or bradykinin B(1) receptor antagonist, R-954 were given intraperitoneally (100 microg/kg), 30 min before induction.. In allergic inflammation, pre-treatment with R-954 reduced eosinophil infiltration into the lungs, mucus secretion and the airway hyperreactivity to methacholine. Pre-treatment with HOE-140 increased eosinophil infiltration but did not affect the other parameters. In immune-complex inflammation, HOE-140 increased neutrophil infiltration but not their activation nor the hemorrhagic lesions. R-594 pre-treatment did not change the parameters examined.. These results show important modulatory effects of bradykinin B(1) and B(2) receptor antagonists in both models of lung inflammation.

Topics: Animals; Bradykinin; Bradykinin B1 Receptor Antagonists; Bradykinin B2 Receptor Antagonists; Bradykinin Receptor Antagonists; Bronchoconstriction; Eosinophils; Hypersensitivity; Immune Complex Diseases; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Pneumonia

IL-12 consists of two disulfide-linked subunits, p40 and p35, that form functionally active heterodimers for the induction of Th1 cells. In contrast to IL-12 heterodimers, p40 monomers and homodimers possess inhibitory effects on Th1 cells leading to the creation of a Th2 environment. Although it has been shown that IL-12p40 acts as antagonist of IL-12p70 in vitro, no evidence is currently available whether IL-12p40 is functional in vivo. We now report that IL-12p40 plays an important pathological role in anintestinal allergic disease. A high expression of IL-12p40 protein was demonstrated in epithelial cells, dendritic cells, and macrophages in large but not small intestine of allergic diarrhea-induced mice. Interestingly, neutralization with anti-IL-12p40 mAbs reduced the incidence and delayed the onset of disease development. Lower levels of ovalbumin (OVA)-specific IgE Abs in serum were detected in anti-IL-12p40 mAb-treated mice than in control Ab-treated mice. The secretion of Th2 cytokines and eotaxin by the mononuclear cells isolated from the large intestine of anti-IL-12p40 mAb-treated mice was significantly decreased. Finally, the removal of the IL-12p40 gene resulted in complete inhibition of disease development. These results show that over-expression of IL-12p40 is an important contributing factor for the generation of the dominant Th2-type environment in the large intestine of mice with allergic diarrhea.

Topics: Animals; Antigens, Neoplasm; Blotting, Western; Chemokine CCL11; Chemokines, CC; Cytokines; Dendritic Cells; Diarrhea; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Hypersensitivity; Immunoglobulin E; Immunohistochemistry; Interleukin-12; Interleukin-12 Subunit p40; Intestine, Large; Macrophages; Mice; Mice, Knockout; Mucins; Ovalbumin; Protein Subunits; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Th2 Cells

Bronchoconstrictor responses were measured in lungs isolated from spontaneously hypertensive (SHR) and normotensive rats, perfused via the airways. Lungs from SHRs were more responsive than lungs from normotensive rats to methacholine, 5-hydroxytryptamine (5-HT), arachidonic acid or prostaglandin H(2). The responses of SHR airways to methacholine or 5-HT were unaffected by pretreatment in vivo with an inhibitor of nitric oxide (NO) synthase, N(omega)-nitro-L-arginine methyl ester hydrochloride (L-NAME, 30 mg kg(-1)), although responses in normotensive airways to methacholine, but not to 5-HT, were enhanced. Antigen challenge of isolated lungs from actively sensitized rats elicited bronchoconstriction, not different between strains. Pretreatment with L-NAME increased the response to antigen challenge only in normotensive lungs. Compound 48/80 induced bronchoconstriction in lungs from either strain, equally. These responses to compound 48/80 were unaffected by L-NAME pretreatment. Thus, SHR airways lack relaxing factors and degranulation of mast cells in SHR lungs was not affected by endogenous NO.

Topics: Animals; Bronchial Hyperreactivity; Bronchoconstriction; Bronchoconstrictor Agents; Cell Degranulation; Dose-Response Relationship, Drug; Enzyme Inhibitors; Hypersensitivity; In Vitro Techniques; Lung; Male; Mast Cells; Methacholine Chloride; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Ovalbumin; p-Methoxy-N-methylphenethylamine; Rats; Rats, Inbred SHR; Rats, Wistar

Our previous study has shown that Chlamydia lung infection can inhibit local eosinophilic inflammation induced by allergen sensitization and challenge, which is correlated with altered cytokine production. In the present study, we examined the role played by dendritic cells (DC) in chlamydial infection-mediated modulation of allergic responses. The results showed that DC freshly isolated from Chlamydia-infected mice (iIDC), unlike those from naive control mice (iNDC), could efficiently modulate immune responses to ovalbumin in vitro and in vivo. Co-culture of freshly isolated DC with naive CD4 cells from T cell receptor transgenic mice (DO11.10) showed that iIDC directed Th1-dominant, while iNDC directed Th2-dominant, allergen-specific CD4 T cell responses. Moreover, adoptive transfer of iIDC, but not iNDC, could inhibit systemic and local eosinophilia induced by allergen exposure. The reduction of eosinophilia was associated with a decrease in IL-5 receptor expression on bone marrow cells and the production of IL-5 and IL-13 by T lymphocytes. Analysis of the DC showed that iIDC expressed significantly higher levels of mRNA for Toll-like receptor 9 and produced more IL-12 compared to iNDC. The data demonstrate a critical role played by DC in infection-mediated inhibition of allergic responses.

Topics: Animals; CD4-Positive T-Lymphocytes; Cell Differentiation; Cells, Cultured; Chlamydia; Chlamydia Infections; Coculture Techniques; Dendritic Cells; Eosinophilia; Hypersensitivity; Immunohistochemistry; Interleukin-13; Interleukin-5; Lymphocyte Activation; Membrane Glycoproteins; Mice; Mice, Transgenic; Ovalbumin; Receptors, Cell Surface; Reverse Transcriptase Polymerase Chain Reaction; Toll-Like Receptors

Nasal administration of major peptide T cell epitopes gives contradictory data on the induction of peripheral tolerance.. To compare the prophylactic effect of intranasal treatment (INT) on the development of an allergic response, using either ovalbumin (OVA) or its major T cell epitope OVA 323-339 (OVAp).. BALB/c mice were treated intranasally with OVA or OVAp and subsequently immunized s.c. with OVA. Anti-OVA-specific antibody, T cell proliferation and cytokine responses were analysed. In an adoptive transfer model using OVAp specific TCR transgenic (Tg) T cells from D011.10 mice, in vivo tracking and characterization of transferred T cells in the cervical, inguinal and bronchial lymph nodes (BLN) and in the spleen were determined by FACS analysis.. Prophylactic INT with OVA induced T cell tolerance towards subsequent OVA s.c. immunizations, inhibiting OVA specific T cell proliferation, IgE and IgG1 production, in contrast to INT with OVAp, which was unable to induce tolerance. In vivo analysis of transferred OVA-specific TCR Tg T cells showed that INT with OVA induced a preferential activation of T cells in BLN, as opposed to a broad, systemic activation with OVAp. In vivo, OVAp INT led to faster and more sustained cell division cycles than OVA INT. Ex vivo, tolerance to OVA was associated with the generation of IL-10 secreting CD4(+) T cells in BLN of OVA-treated mice only.. INT with OVA but not with OVAp led to regional (as opposed to systemic) T cell activation and the induction of IL-10 secreting CD4(+) T cells in BLN, potentially critical steps in the induction of T cell-specific tolerance via the nasal route.

Topics: Administration, Intranasal; Adoptive Transfer; Animals; CD4-Positive T-Lymphocytes; Cell Division; Epitopes, T-Lymphocyte; Female; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Interleukin-10; Lymphocyte Activation; Lymphoid Tissue; Mice; Mice, Inbred BALB C; Ovalbumin; Peptide Fragments; T-Lymphocyte Subsets

High genetic risk (HR) of atopy among unstratified populations of infants is associated with attenuated IFN-gamma responses. However, the role of IFN-gamma in progression from HR status to active disease is less clear.. To identify immune function markers in neonates with HR that are associated with positive atopic outcomes at 2 years.. Cord blood mononuclear cells (CBMCs) were collected from 175 children with HR and cryopreserved. The children were assessed for atopy by skin prick at 0.5 and 2 years. CBMCs were thawed and stimulated with allergens and mitogens PHA and staphylococcal enterotoxin B (SEB), and cytokine responses were determined.. No correlations were observed between allergen-specific CBMC responses and atopic outcomes. In contrast, sensitization was strongly associated with polyclonal IFN-gamma responses to both PHA (P=.002) and SEB (P=.005), and also with SEB-induced IL-5 (P =.05), IL-10 (P =.02), and IL-13 (P =.01). Logistic regression analysis identified elevated PHA-induced IFN-gamma and SEB-induced IL-13 responses as the strongest independent predictors of atopy development. Cell separation studies confirmed CD8+ T cells as the source of approximately 90% of IFN-gamma production.. IFN-gamma produced by CD8+ T cells may synergize with T(H)2 cytokines in driving atopy development in children with HR.

Topics: Allergens; CD8-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Fetal Blood; Genetic Predisposition to Disease; Humans; Hypersensitivity; Immunization; Infant, Newborn; Interferon-gamma; Monocytes; Ovalbumin; Pyroglyphidae

Respiratory syncytial virus (RSV) infection has been shown to be a risk factor for the development of allergy in humans and mice. The allergy-enhancing properties of RSV may be dependent on atopic background and an individual's history of RSV infection. We examined the influence of the timing of infection and prior inoculation with RSV in a mouse model of allergic asthma. Mice were sensitized to and challenged with ovalbumin (OVA) and were inoculated with RSV either before or during the sensitization or challenge period. One group of mice was inoculated with RSV both before sensitization to OVA and during challenge with OVA. Increased pulmonary expression of interleukin (IL)-4, IL-5, and IL-13 mRNA and aggravated alveolitis and hypertrophy of mucus-producing cells were observed only when OVA-sensitized mice were inoculated with RSV shortly before or during challenge with OVA. Despite protection against viral replication, prior inoculation with RSV did not abrogate RSV-enhanced, OVA-induced expression of T helper 2 (Th2) cytokines in the lung. In conclusion, inoculation with RSV enhances allergic disease only when the immune system has already been Th2-primed by the allergen (i.e., OVA). This RSV-enhanced allergy is not completely abrogated by prior inoculation with RSV.

Topics: Animals; Asthma; Female; Histocytochemistry; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; Specific Pathogen-Free Organisms; Statistics, Nonparametric

Histamine-1-receptor (H1R)-antagonists were shown to influence various immunological functions on different cell types and may thus be employed for immune-modulating strategies for the prevention of primary immune responses.. The aim of this study was to investigate the effects of an H1R-antagonist on allergen-induced sensitization, airway inflammation (AI) and airway hyper-reactivity (AHR) in a murine model.. BALB/c mice were systemically sensitized with ovalbumin (OVA) (six times, days 1-14) and challenged with aerosolized allergen (days 28-30). One day prior to the first and 2 h prior to every following sensitization, mice received either 1 or 0.01 microg of desloratadine (DL) or placebo per os.. Sensitization with OVA significantly increased specific and total IgE and IgG1 serum levels, as well as in vitro IL-5 and IL-4 production by spleen and peribronchial lymph node (PBLN) cells. Sensitized and challenged mice showed a marked eosinophilic infiltration in broncho-alveolar lavage fluids and lung tissues, and developed in vivo AHR to inhaled methacholine. Oral treatment with DL prior to OVA sensitization significantly decreased production of OVA-specific IgG1, as well as in vitro Th2-cytokine production by spleen and PBLN cells, compared with OVA-sensitized mice. Moreover, eosinophilic inflammation and development of in vivo AHR were significantly reduced in DL-treated mice, compared with sensitized controls.. Treatment with H1R-anatagonist prior to and during sensitization suppressed allergen-induced Th2 responses, as well as development of eosinophilic AI and AHR. This underscores an important immune modulating function of histamine, and implies a potential role of H1R-anatagonists in preventive strategies against allergic diseases.

Topics: Administration, Oral; Allergens; Animals; Bronchial Hyperreactivity; Cells, Cultured; Female; Histamine H1 Antagonists; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Interleukin-5; Loratadine; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Spleen

Topics: Animals; Asthma; Humans; Hypersensitivity; Immunization; Influenza, Human; Orthomyxoviridae; Ovalbumin; Virus Diseases

When wild-type BALB/c mice were transferred with OVA-specific Th2 cells followed by OVA inhalation, a severe eosinophilia, mucus hypersecretion and airway hyper-responsiveness (AHR) was induced in parallel with a marked elevation of IL-4, IL-5 and IL-13 levels in bronchoalveolar lavage fluid (BALF). However, neither eosinophilia, AHR nor mucus hypersecretion was induced in Th2 cell-transferred STAT6-/- mice. The failure of eosinophilia was not due to the defect of Th2 cytokine production in BALF of STAT6-/- mice transferred with Th2 cells, but because of the defect of STAT6-dependent eotaxin production. Indeed, intranasal administration of eotaxin reconstituted pulmonary eosinophilia but not AHR and mucus hypersecretion in OVA-inhalated STAT6-/- mice. These results initially provided direct evidence that STAT6-dependent eotaxin production is essential for pulmonary eosinophilia. We also dissociated the role of STAT6 for eosinophilia from that for AHR and mucus hypersecretion. Thus, STAT6 also plays a critical role at late phase of Th2-dependent allergy induction.

Topics: Administration, Intranasal; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Differentiation; Chemokine CCL11; Chemokines, CC; Eosinophilia; Hypersensitivity; Lung; Mice; Mucus; Ovalbumin; Recombinant Proteins; STAT6 Transcription Factor; Th2 Cells; Trans-Activators

Topics: Adjuvants, Pharmaceutic; Allergens; Animals; Antibody Formation; Benzalkonium Compounds; Denmark; Drug Interactions; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Helper-Inducer; Tetraethylammonium; Th2 Cells

Particle exposure has traditionally been monitored as mass concentration of PM10 (particles with an aerodynamic diameter less than 10 microm), more recently also as PM2.5. The mass concentration is strongly influenced by the large particles. Therefore, particle mass is a poor measure for characterizing the amount of the small, possibly more biologically potent particles. We used polystyrene particles (PSP) ranging in diameter from 0.0588 to 11.14 microm, carbon black (CB), and diesel exhaust particles (DEP), to study the adjuvant effect of particles on the immune response to the allergen ovalbumin (OVA) after sc injection into the footpad of BALB/cA mice. At a given mass dose, the small particles (0.0588 and 0.202 microm PSP, CB, and DEP) increased the allergen-specific IgE serum levels to a substantially higher degree than the larger particles (1.053, 4.64, and 11.14 microm PSP). Further, in the draining lymph node during the primary response, the fine particles (0.202 microm) with OVA increased cell numbers, expression of surface markers (CD19, MHC class II, CD86, and CD23) and ex vivo production of IL-4 and IL-10, whereas the largest (11.14 microm) particles did not. Linear regression analyses indicated that the IgE response was not predicted by particle mass (R2 = 0.06), but was predicted by the total particle surface area (R2 = 0.64), number of particles (R2 = 0.62), and particle diameter (R2 = 0.58). In conclusion, we found that fine particles exerted stronger adjuvant effects on allergic responses than larger particles at equal mass doses. Consequently, the dose described as total particle surface area or particle number predicts the adjuvant effect of particles better than the currently used particle mass.

Topics: Adjuvants, Immunologic; Animals; Antigens, Surface; Carbon; Cytokines; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lymph Nodes; Lymphocyte Count; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Phenotype; Polystyrenes; Predictive Value of Tests; Regression Analysis; Surface Properties; Thymidine; Vehicle Emissions

There is growing interest in the immune-stimulating effect and in particular, the anti-allergic effect, of lactic acid bacteria (LAB). However, no comprehensive studies have been done that compare the immune-stimulating potential of LAB strains.. The in vitro immune-stimulating effects on Th1/Th2 balance of more than 100 LAB strains were compared in splenocytes from ovalbumin-sensitized Th2-polarized mice. The in vivo anti-allergic ability of strain KW3110 was studied in the Th2-polarized model by detecting serum IgE concentration, Th1/Th2 cytokine secretion from splenocytes, and the expression of co-stimulatory molecules on macrophages.. In vitro studies from Th2-polarized splenocytes, using IL-12 as a Th1 parameter and IL-4 secretion as a Th2 parameter revealed a wide variety of IL-12-inducing and IL-4-repressing activities, depending on the strain of LAB, not depending on the species. However, evaluation of individual strains in vivo revealed that after exposure to Lactobacillus paracasei KW3110 strain, the serum IgE elevation elicited by repeated OVA injection of mice was strongly inhibited. Cytokine secretion from splenocytes 20 weeks after KW3110 administration showed increased IL-12 and decreased IL-4 expression. Both CD40 and B7-1 expression on macrophages was upregulated by administration of KW3110.. Improving the consequences of the Th1/Th2 imbalance by administration of LAB was dependent upon the LAB strain rather than the LAB species. Oral KW3110 administration in the mouse allergy model directed the Th1/Th2 balance toward Th1 through the maturation of APCs and inhibition of serum IgE elevation.

Topics: Animals; B7-1 Antigen; CD40 Antigens; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Interleukin-12; Interleukin-4; Lactobacillus; Macrophages, Peritoneal; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Spleen; Th1 Cells; Th2 Cells; Up-Regulation

IgE is critical in the pathogenesis of allergic disorders. In this report, we investigated the differential regulation of antigen-specific and by-stander IgE. Ovalbumin (OVA) immunization did not increase IgE producing cells in the spleen, but significantly enhanced the intracellular IgE content of all IgE+ cells. In contrast, OVA induced a significant increase of IgE+ cells in the draining lymph nodes (LN). Furthermore, OVA-specific IgE was detected only in the ex vivo cultures of the draining LN but not the spleen cells, while total IgE was increased in both cultures. These results indicated that antigen-specific IgE was mainly produced in the draining LN, while the spleen was a major source for by-stander IgE. Anti-IL-4, but not anti-IL-13, antibody blocked the expansion of IgE producing cells in the draining LN as well as systemic OVA-specific and total IgE levels, indicating IL-4 was important in both antigen-specific IgE generation and total IgE upregulation.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Flow Cytometry; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-13; Interleukin-4; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Specific Pathogen-Free Organisms; Spleen; Up-Regulation

Exposure to moulds in indoor air is thought to induce asthma in susceptible persons. Moulds may contain several potent allergens. However, more importantly, moulds may increase the allergic response to other allergens (adjuvant effect). Previously, we have found that a beta-1,3-glucan from the cell wall of the fungus Sclerotinia sclerotiorum increases the allergic response to the model allergen ovalbumin (OVA) in a mouse model.. In the present study, we wanted to confirm the adjuvant effect of another beta-1,3-glucan, MacroGard (MG) from baker's yeast in this model. More importantly, we wished to explore the putative effects of extracts from the moulds Cladosporium herbarum (CH) and Penicillium chrysogenum (PC) using the very same model as used to explore effects of beta-glucans.. Groups of eight Balb/c mice were injected with OVA alone, OVA+extract or OVA+MG, into one footpad. On day 21, all mice were reinjected with OVA, before exsanguination on day 26. The levels of OVA-specific IgE, IgG1 and IgG2a in serum were measured by ELISA.. Compared with OVA alone, OVA+MG, OVA+CH extract and OVA+PC extract increased OVA-specific IgE and IgG1 levels significantly. For all groups, the levels of IgG2a anti-OVA remained similar to those of the OVA-alone group.. Our results show that extracts from CH and PC, and the beta-1,3/1,6-glucan from baker's yeast have adjuvant effects on the allergic response in mice.

Topics: Adjuvants, Immunologic; Animals; beta-Glucans; Biological Factors; Cladosporium; Endotoxins; Female; Fungal Proteins; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Penicillium chrysogenum; Saccharomyces cerevisiae

In mice, intratracheal challenges with antigen (ovalbumin) or recombinant murine interleukin-13 (IL-13) induce lung inflammation, bronchial hyperreactivity (BHR), and mucus accumulation as independent events (Singer M, Lefort J, and Vargaftig BB. Am J Respir Cell Mol Biol 26: 74-84, 2002), largely mediated by leukotrienes (LT). We previously showed that LTC(4) was released 15 min after ovalbumin, and we show that it induces the expression of monocyte chemoattractant proteins 1 and 5 and KC in the lungs, as well as IL-13 mRNA. Instilled intratracheally, these chemokines induced BHR and mucus accumulation, which were inhibited by the 5-lipoxygenase inhibitor zileuton and by the cysteinyl-LT receptor antagonist MK-571, suggesting mediation by cysteinyl-LT. Because these chemokines also induced release of LT into the bronchoalveolar lavage fluid and IL-13 into the lungs, we hypothesize that LT- and chemokine-based loops for positive-feedback regulations cooperate to maintain and amplify BHR and lung mucus accumulation after allergic challenge and in situations where IL-13, LT, or chemokines are generated.

Topics: Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemokines; Hypersensitivity; Interleukin-13; Leukotriene C4; Leukotrienes; Lung; Male; Metaplasia; Mice; Mice, Inbred Strains; Mucins; Mucus; Ovalbumin; Recombinant Proteins; Respiratory Mucosa; RNA, Messenger

In mice, respiratory syncytial virus (RSV) infection during allergic provocation aggravates the allergic Th2 immune response, characterised by production of interleukin (IL)-4, IL-5, and IL-13, and eosinophilic inflammation. This enhancement of the Th2 response occurs simultaneously with a strong RSV-induced Th1 cytokine response (IL-12 and IFN-gamma). The present study investigated whether IFN-gamma and IL-12 are critically involved in this RSV-enhanced OVA allergy. Therefore, IFN-gammaR- and IL-12-deficient mice (both on a 129/Sv/Ev background) were sensitised and challenged with ovalbumin (OVA) and infected with RSV during the OVA challenge period. Neither gene deletion affected the development of ovalbumin-induced allergic inflammation in mice. However, when OVA-allergic IFN-gammaR deficient mice were infected with RSV, an increased pulmonary eosinophilic infiltrate and increased IL-4 and IL-13 mRNA expression in lung tissue were observed compared with identically treated wild-type mice. In contrast, deficiency of IL-12 did not aggravate the Th2 immune and inflammatory response in OVA/RSV-treated mice, compared with wild-type. In conclusion, the virus-induced IFN-gamma response diminishes the Th2 inflammatory response during OVA allergy but fails to prevent totally the enhancement of the OVA allergy by RSV. In contrast, IL-12 is not involved in inhibiting nor increasing the RSV-enhanced allergy in 129/Sv/Ev mice.

Topics: Animals; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunization; Immunoglobulin E; Interferon-gamma; Mice; Mice, Knockout; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; RNA, Messenger

Coupling of ovalbumin (OVA) to anti-DEC-205 monoclonal antibody (mAb) (alphaDEC) induced the proliferation of OVA-specific T cells in vivo. Expansion was short-lived, caused by dendritic cells (DCs), and rendered T cells anergic thereafter. Phenotypic analysis revealed the induction of CD25+/CTLA-4+ T cells suppressing proliferation and interleukin-2 (IL-2) production of effector CD4+ T cells. The findings were supported by 2 disease models: (1) CD4+ T-cell-mediated hypersensitivity reactions were suppressed by the injection of alphaDEC-OVA and (2) the application of hapten-coupled alphaDEC-205 reduced CD8+ T-cell-mediated allergic reactions. Thus, targeting of antigens to immature DCs through alphaDEC antibodies led to the induction of regulatory T cells, providing the basis for novel strategies to induce regulatory T cells in vivo.

Topics: Animals; Antibodies, Monoclonal; Antigen Presentation; Antigens; Antigens, CD; Antigens, Differentiation; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; CTLA-4 Antigen; Dendritic Cells; Dermatitis, Contact; Haptens; Hypersensitivity; Hypersensitivity, Delayed; Interleukin-2; Lymphocyte Activation; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Interleukin-2; T-Lymphocytes

Recognition of foreign substances by innate immunity through pattern recognition receptors (PRRs) regulates acquired immunity such as allergic reaction. Because PRRs recognize heterogeneous ligands, daily food intake can potentially regulate immune allergic reaction.. Elucidation of the effect of lambda-carrageenan on allergic reactions was aimed.. IFN-gamma and IL-4 was measured in in vitro T cell-stimulated culture. Cytokine production from macrophages in response to lambda-carrageenan was measured as indicator for innate immunity activation. Mice were immunized with OVA in alum to induce specific IgE, and then histamine release was induced by systemic injection of OVA.. Activation of innate immunity by lambda-carrageenan is dependent on Toll-like receptor-4 (TLR4) and MyD88, in which induction of pro-inflammatory cytokines such as TNF-alpha and IL-6 was largely impaired in macrophages from TLR4- and MyD88-deficient mice. Footpad oedema, a model for in vivo inflammatory reactions, was significantly reduced in these mice. Similar to recent evidence showing a preference for the stimulation of Th1 via TLR/MyD88 signalling, lambda-carrageenan showed enhanced IFN-gamma and decreased IL-4 in stimulated T cell cultures. Interestingly, increased IFN-gamma production was still seen in TLR4- and MyD88-deficient splenocytes. Oral administration of lambda-carrageenan to immunized mice successfully decreased OVA-specific IgE, and lambda-carrageenan was also effective in previously immunized mice. Further, serum histamine release upon systemic challenge of OVA was significantly inhibited. Neither OVA-specific IgG1/IgG2a nor cytokine secretion from in vitro cultures were altered, suggesting the involvement of multiple PRRs as demonstrated by TLR4/MyD88-independent IFN-gamma up-regulation. The simultaneous feeding of OVA with lipopolysaccharide abrogated oral tolerance, but lambda-carrageenan was not only devoid of such an effect but was also found to promote oral tolerance in the absence of TLR4.. lambda-Carrageenan was suggested to be a useful dietary supplement to ameliorate allergic reactions while maintaining oral tolerance-dependent intestinal homeostasis.

Topics: Adaptor Proteins, Signal Transducing; Administration, Oral; Animals; Antigens, Differentiation; Carrageenan; Cells, Cultured; Drosophila Proteins; Edema; Female; Histamine Release; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Interferon-gamma; Interleukin-4; Macrophage Activation; Membrane Glycoproteins; Mice; Mice, Inbred Strains; Mitogens; Myeloid Differentiation Factor 88; Ovalbumin; Receptors, Cell Surface; Receptors, Immunologic; Spleen; Th1 Cells; Th2 Cells; Toll-Like Receptor 4; Toll-Like Receptors

T cell priming, as determined by allergen-induced proliferative responses, is believed to occur principally in early childhood in both atopic and non-atopic infants under the influence of multiple factors including environmental allergen exposure. It is considered that T cell priming with expansion of Th2 cells is a crucial factor in the development of atopic disease.. To examine T cell priming to commonly encountered allergens in childhood in relation to age.. In a cross-sectional study T cell proliferation in relation to age was examined for three common allergens, ovalbumin (OVA), house dust mite (HDM) and rye grass pollen (RYE), in atopic and non-atopic children. The effect of age on Th1 (IFN-gamma) and Th2 (IL-5 and IL-13) cytokine production in response to these allergens was investigated to examine the possibility of immune deviation with time.. A significant increase in T cell proliferation with age was observed with RYE among atopic children only. However, the same was not observed with the two other allergens studied (i.e. OVA and HDM). In addition, RYE-induced (but not HDM or OVA) cytokine production showed an increased Th2 deviation with age as reflected in the increasing IL-5/IFN-gamma and IL-13/IFN-gamma ratios only among the atopic subjects with rye grass pollen sensitivity.. These findings suggest that grass pollen sensitivity in childhood is accompanied by a progressive accumulation of allergen-primed T cells and progressive deviation of the allergen-induced cytokine response towards a Th2 response in atopic subjects throughout childhood.

Topics: Adolescent; Age Factors; Allergens; Animals; Cells, Cultured; Child; Child, Preschool; Cross-Sectional Studies; Cytokines; Enzyme-Linked Immunosorbent Assay; Humans; Hypersensitivity; Immunoglobulin E; Infant; Lolium; Mites; Ovalbumin; Pollen; Regression Analysis; T-Lymphocytes; Th2 Cells

We investigated the development of airway hyperreactivity (AHR) and inflammation in the lungs of nine genetically diverse inbred strains of mice [129/SvIm, A/J, BALB/cJ, BTBR+(T)/tf/tf, CAST/Ei, C3H/HeJ, C57BL/6J, DBA/2J, and FVB/NJ] after sensitization and challenge with ovalbumin (OVA). At 24, 48, and 72 h post-OVA exposure, the severity of AHR and eosinophilic inflammation of the mouse strains ranged from relatively unresponsive to responsive. The severity of the airway eosinophilia of some strains did not clearly correlate with the development of AHR. The temporal presence of T helper type 2 cytokines in lung lavage fluid also varied markedly among the strains. The levels of IL-4 and IL-13 were generally increased in the strains with the highest airway eosinophilia at 24 and 72 h postexposure, respectively; the levels of IL-5 were significantly increased in most of the strains with airway inflammation over the 72-h time period. The differences of physiological and biological responses among the inbred mouse strains after OVA sensitization and challenge support the hypothesis that genetic factors contribute, in part, to the development of allergen-induced airway disease.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Eosinophils; Hypersensitivity; Interleukin-13; Interleukin-4; Interleukin-5; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred DBA; Neutrophils; Ovalbumin; Species Specificity; Th2 Cells

The immediate responses of the upper respiratory tract (URT) to the irritants acrolein and acetic acid were examined in healthy and allergic airway-diseased C57Bl/6J mice. Acrolein (1.1 ppm) and acetic acid (330 ppm) vapors induced an immediate increase in flow resistance, as measured in the surgically isolated URT of urethane-anesthetized healthy animals. Acrolein, but not acetic acid, induced a small URT vasodilatory response. In awake spontaneously breathing mice, both vapors induced a prolonged pause at the start of expiration (a response mediated via stimulation of nasal trigeminal nerves) and an increase in total respiratory specific airway flow resistance, the magnitude of which was similar to that observed in the isolated URT. Both responses were significantly reduced in animals pretreated with large doses of capsaicin to defunctionalize sensory nerves, strongly suggesting a role for sensory nerves in development of these responses. The breathing pattern and/or obstructive responses were enhanced in mice with ovalbumin-induced allergic airway disease. These results suggest that the primary responses to acrolein and acetic acid vapors are altered breathing patterns and airway obstruction, that sensory nerves play an important role in these responses, and that these responses are enhanced in animals with allergic airway disease.

Topics: Acetic Acid; Acrolein; Administration, Inhalation; Airway Obstruction; Airway Resistance; Animals; Female; Hypersensitivity; Irritants; Male; Mice; Mice, Inbred C57BL; Neurons, Afferent; Ovalbumin; Pulmonary Ventilation; Respiration; Respiratory Mechanics; Respiratory Physiological Phenomena; Respiratory System; Respiratory Tract Diseases; Vasodilation

Vaccination with the antiallergic drug Histaglobin is used to treat a broad range of human allergic diseases including bronchial asthma, allergic rhinitis, and atopic dermatitis. In order to further elucidate its functional activity, Histaglobin was investigated in an in vivo mouse allergy model. Mice were sensitized with ovalbumin either prior to or after Histaglobin treatment, and its antiallergic potential was evaluated. Ovalbumin-sensitized mice exhibited increased serum levels of IL-4, tumor necrosis factor alpha (TNF-alpha), and an increase of total and ovalbumin-specific IgE; total and ovalbumin-specific IgG levels were also elevated. Subsequent administration (therapeutic treatment) of Histaglobin resulted in a decrease of total and specific serum IgE levels; total and specific IgG1 serum levels were reduced by more than 50% and 45%, respectively; the mice displayed a down-regulation of IL-4 and TNF-alpha serum levels and showed increased levels of IFN-gamma and IgG2a. Mice pretreated with Histaglobin, prior to ovalbumin sensitization (prophylactic treatment), were found to be widely unresponsive to ovalbumin. They exhibited higher serum levels of IFN-gamma and IgG2a (total and specific) compared to saline-treated control mice. The inhibitory effects were still observed 1 month post-immunization. Our data, indicating a Histaglobin-induced modulation of the Th1/Th2 balance in favour of Th1, correspond with the well-known antiallergic activity of Histaglobin observed in patients.

Topics: Animals; Anti-Allergic Agents; Disease Models, Animal; Drug Combinations; Female; gamma-Globulins; Histamine; Hypersensitivity; Immunoglobulin E; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha

Oral administration of allergens can induce immune tolerance to specific allergens in rodents and hence might be a possibility to prevent and treat allergic diseases in human subjects. However, the gastrointestinal tract of mice is different from that of human subjects. The absorption of specific antigens and subsequent antigen presentation to intestinal T cells is different in both species, making it difficult to extrapolate results.. We investigated primary oral tolerance to ovalbumin (OVA) in an IgE high-responder dog model, which is more predictive for human allergic diseases than corresponding rodent models.. Oral tolerance was induced by means of a 28-day treatment with OVA dissolved in cow's milk.. We observed reduced OVA-specific IgE and IgG production in response to ensuing subcutaneous challenges. Allergic conjunctivitis induced by means of ocular and airway provocation was significantly reduced in tolerized animals compared with that seen in nontolerized control animals. In addition, eosinophilia and neutrophilia in bronchoalveolar lavage fluid and bronchoconstriction after airway allergen challenge were significantly suppressed in tolerized animals. Cytokine analysis by means of real-time PCR on bronchoalveloar fluid cells after allergen challenge revealed a high-level expression of IL-10 and transforming growth factor beta, predominantly in the CD14(+) population.. Feeding infant beagles with OVA for 4 weeks is sufficient to prevent hallmark manifestations of asthma and allergy in adult life. The mechanism of oral tolerance involved an increased expression of IL-10 and transforming growth factor beta cytokines.

Topics: Administration, Oral; Animals; Asthma; Conjunctivitis; Dogs; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Interleukin-10; Ovalbumin; RNA, Messenger; Transforming Growth Factor beta

Conventional models of allergic airway inflammation involve intraperitoneal administration of ovalbumin in conjunction with a chemical adjuvant (generally aluminum hydroxide) to generate allergic airways inflammation. Here we have investigated the effect of respiratory mucosal exposure to a ragweed extract in the absence of chemical adjuvant on the generation of allergic responses.. We sought to develop a mouse model of ragweed-induced allergic airway inflammation through mucosal sensitization and to investigate the role of GM-CSF in this process.. Ragweed was delivered intranasally to an airway microenvironment enriched with GM-CSF, which was achieved by means of either multiple coadministrations of recombinant GM-CSF or a single delivery of an adenoviral vector carrying the GM-CSF transgene.. Administration of a purified ragweed extract leads to T(H)2 sensitization (and not inhalation tolerance) accompanied by mild airway inflammation, modest clinical symptoms, and moderate production of T(H)2 cytokines by splenocytes on ragweed restimulation. The administration of anti-GM-CSF antibodies in conjunction with ragweed diminished T(H)2-associated cytokine production. These responses were amplified by enriching the airway microenvironment with GM-CSF. Under these conditions, all T(H)2-associated immune-inflammatory responses, as well as the clinical responses, were considerably enhanced. To investigate the mechanism underlying these effects, we examined lung mononuclear cells by means of flow cytometry and detected a substantial expansion of antigen-presenting cells, particularly dendritic cells, as well as a substantially increased activation of these antigen-presenting cells, as demonstrated by the expression of B7 molecules, particularly B7.2.. GM-CSF plays an important role in the generation of allergic immune-inflammatory responses to ragweed.

Topics: Ambrosia; Animals; Asthma; Cytokines; Female; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Recombinant Proteins; Th2 Cells

Dehulled adlay is known as a natural Chinese medicine having antiallergic activity, although its mechanism remains unclear. This study examined the effects of dehulled adlay on antigen-specific antibody and cytokine production. Mice were immunized three times with ovalbumin (OVA) in alum adjuvant. It was found that oral administration of dehulled adlay in mice suppressed the production of IgE against OVA antigen. Serum anti-OVA IgG(2a) antibody levels were significantly increased in mice after oral administration of dehulled adlay. Furthermore, the production of IL-2 by OVA-stimulated splenocytes was augmented in dehulled adlay-fed mice. Although dehulled adlay had no effect on the serum anti-OVA IgG(1) antibody levels, it had a great capacity to reduce IL-5 secretion by means of OVA-stimulated splenocytes. Hydrothermal processes, including steaming and extrusion cooking, did not change the capacity of dehulled adlay to suppress IgE production. Three fractions of dehulled alday, including methanolic extract, warm water extract, and residue, were obtained. The methanolic extract exhibited the greatest capacity to reduce anti-OVA IgE production. These results suggest that dehulled adlay has a modulating ability to shift the balance from Th2 to Th1 dominance in the T cell mediated immune system and may be beneficial for the treatment of allergic disorders.

Topics: Animals; Coix; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Interleukin-5; Male; Methanol; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Th1 Cells; Th2 Cells

The role of platelets in inflammation is recognized but poorly characterized, and little is known of their interaction with leukocytes. However, platelet-leukocyte interactions have been demonstrated in cardiovascular disease, culminating in enhanced leukocyte recruitment.. This study was undertaken to assess the possibility and potential role of similar phenomena occurring in asthmatic patients, a murine model of allergic inflammation, and in vitro adhesion studies.. Asthmatic patients had blood taken at various time points to document the degree of leukocyte activation and the presence of platelet-leukocyte aggregates through FACS analysis before and after allergen exposure. Similar studies were carried out in mice exposed to allergen after previous sensitization, with some groups being selectively depleted of platelets through both an immunologic (antiplatelet antiserum) and nonimmunologic (busulfan) method. Additionally, lavage fluid and airway tissue were analyzed to assess the degree of pulmonary leukocyte recruitment. The importance of platelets on leukocyte adhesion to the endothelium was then assessed with in vitro incubation of radiolabeled leukocytes in the presence of activated platelets on cultured human vascular endothelial cells.. We have observed circulating platelet-leukocyte aggregates in the blood of allergic asthmatic patients during the allergen-induced late asthmatic response and in sensitized mice after allergen exposure. In platelet-depleted mice infiltration of leukocytes into airways after allergen challenge was significantly reduced and could be restored by means of infusion of platelets from allergic animals, indicating an essential role for platelets in leukocyte recruitment. CD11b expression on leukocytes involved in aggregates with platelets, although not on free leukocytes, was upregulated. Furthermore, the presence of autologous platelets augmented the adhesion of human polymorphonuclear leukocytes to cultured vascular endothelial cells, an effect that was found to be endothelial cell dependent and to involve platelet activation.. These results suggest that platelet participation in cell recruitment occurs at the level of the circulation and might involve the priming of leukocytes for subsequent adhesion and transmigration into tissues.

Topics: Adult; Allergens; Animals; Blood Platelets; CD11b Antigen; Cell Adhesion; Cell Movement; Endothelium, Vascular; Female; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Leukocytes; Male; Mice; Mice, Inbred C57BL; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Ovalbumin; Peroxidase

1. We investigated the mediators responsible for mechanical hypersensitivity induced by antigen challenge in rats immunised with ovalbumin (OVA). 2. Challenge with OVA (12.5-100 micro g, intraplantar) caused a dose- and time-dependent mechanical hypersensitivity, which peaked 3 h after, decreased thereafter and reached control levels 24 h later. 3. Levels of TNFalpha, IL-1beta and cytokine-induced neutrophil chemoattractant 1 (CINC-1) were increased in paw skin after antigen challenge. 4. OVA-evoked hypersensitivity was partially inhibited (about 51%) by pretreatment with anti-TNFalpha, IL-1beta and IL-8 sera or with IL-1 receptor antagonist (IL-1ra), but not anti-NGF serum. Pretreatment with thalidomide (45 mg kg(-1)) or pentoxifylline (100 mg kg(-1)) also partially inhibited the hypersensitivity at 1-3 h after challenge. 5. Pretreatment with indomethacin (5 mg kg(-1)) or atenolol (1 mg kg(-1)) reduced the OVA-induced hypersensitivity at 1 and 3 h, but not at 5 h after challenge, while the combination of B(1) and B(2) bradykinin receptor antagonists was ineffective over the same times. 6. Pretreatment with MK886 (5-lipoxygenase-activating protein inhibitor, 3 mg kg(-1)), CP 105696 (LTB(4) receptor antagonist; 3 mg kg(-1)) or dexamethasone (0.5 mg kg(-1)) inhibited the hypersensitivity from 1 to 5 h. Furthermore, LTB(4) levels were increased in the paw skin of challenged rats. 7. In conclusion, our results suggest that the TNFalpha-, IL-1beta- and CINC-1-driven release of prostaglandins, sympathetic amines and LTB(4) mediates the first 3 h of mechanical hypersensitivity induced by antigen challenge in rats. At 5 h after OVA administration, although TNFalpha has some role, LTB(4) is the critical nociceptive mediator.

Topics: Animals; Antigens; Dose-Response Relationship, Drug; Dose-Response Relationship, Immunologic; Hyperalgesia; Hypersensitivity; Injections, Subcutaneous; Leukotriene B4; Male; Ovalbumin; Pain Measurement; Physical Stimulation; Rats; Rats, Wistar; Skin; Up-Regulation

Vitamin D, a common food additive, has been shown to prevent the induction of experimental autoimmune diseases in mice. A possible immune deviation from T(H)1 to T(H)2 responses has been postulated. Although there is no doubt about the beneficial effects of vitamin D, its role in allergy has not been investigated.. To define the role of vitamin D in modulating the development of a T(H)2-mediated disease, we used a murine model of pulmonary eosinophilic inflammation.. Five-week-old mice were primed on day 0 with ovalbumin intraperitoneally. Then they were nasally challenged with ovalbumin on days 7, 8, 9, and 10, and on day 11, samples were studied. Some mice received subcutaneous injections of vitamin D every second day as follows: days -3, -1, 1, 3, 5, 7, and 9. The control groups received PBS on the same days.. Early treatment with vitamin D augmented allergen-induced T-cell proliferation along with T(H)2 cytokine (IL-4 and IL-13) and IgE production. Surprisingly, the local inflammatory response in bronchoalveolar lavage fluid and lung tissue was significantly ameliorated with impaired recruitment of eosinophils and inferior levels of IL-5. These findings were attributed to late treatment with vitamin D after establishment of an early immune response.. We suggest that excess supplementation of vitamin D could influence the development of a sustained T(H)2 response, leading to an increasing prevalence of allergy, whereas vitamin D might hold promising beneficial effects in airway eosinophilia.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophilia; Female; Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; In Vitro Techniques; Interleukin-4; Lung; Mice; Mice, Knockout; Ovalbumin; Th1 Cells; Th2 Cells; Time Factors; Vitamin D

Epidemiological and experimental studies have not only shown that air pollution induces increased pulmonary morbidity, and mortality, but also that air pollution components may potentiate allergic responses. The respiratory allergy model to ovalbumin in the mouse has been shown a useful tool to characterize the adjuvant potency of air pollution components. However, the choice for the most effective route of administration for testing small amounts of air pollution component is hampered by the diversity of routes of administration used. To test the adjuvant activity of airborne particles (Ottawa dust EHC-93), we studied the optimal route of respiratory administration: intranasally (in) and aerosol (aero) in comparison with responses observed by intraperitoneal (ip) with diesel exhaust particles (DEP) as a positive control. Our results show that the combination of in/aero with ovalbumin caused almost similar immunoglobulin (Ig)E and inflammatory responses compared to the ip/aero. In/in application induced less responses for IgE, less inflammation in the lung, and less increased numbers of eosinophils in the bronchoalveolar lavage (BAL). This response increased dramatically when ovalbumin was coadministered with DEP. Subsequently, EHC-93, which is made up of airborne particles, was tested via the in/in route of administration. EHC-93 induced similar IgE responses, inflammation, and eosinophilic response in BAL compared to DEP. In addition, EHC-93 increased the airway responsiveness of the ovalbumin-sensitized mice measured in unrestrained condition and not in nonsensitized control mice. It is concluded that intranasal sensitization with intranasal challenge with airborne particles (EHC-93) is an effective route of administration to show potency of adjuvant activity of airborne particles.

Topics: Administration, Intranasal; Aerosols; Air Pollutants; Animals; Bronchoalveolar Lavage; Disease Models, Animal; Drug Interactions; Dust; Eosinophils; Hypersensitivity; Immunoglobulin E; Inflammation; Infusions, Parenteral; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Vehicle Emissions

In allergic disorders, the role of tumor necrosis factors (TNF) is not well established. We investigated the role of TNF in allergic peritonitis induced by ovalbumin (OVA) challenge in double TNF (TNF-alpha(-/-)/lymphotoxin-alpha(-/-)) knock out (TNF-KO) mice. In the peritoneal lavage of TNF-KO mice, mast cell number and histamine level (radioenzymatic assay) were similar to that in wild type (WT) mice. TNF-alpha (ELISA) and histamine were increased 1 h after challenge in WT mice. However, three days later eosinophil number and eosinophil peroxidase (EPO) levels (colorimetric-enzymatic assay) were found to be lower in TNF-KO mice. A second challenge three days after the first, increased EPO, histamine and IL-6 (ELISA) but did not alter eosinophil and mast cell numbers in both types of mice. On the other hand histamine and IL-6 were higher, while EPO was lower in TNF-KO mice. In conclusion, our findings show that TNF is involved in eosinophil accumulation and inflammatory mediators' release in a murine model of allergy.

Topics: Animals; Disease Models, Animal; Eosinophil Peroxidase; Eosinophils; Histamine; Hypersensitivity; Interleukin-6; Male; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Peritonitis; Peroxidases; Tumor Necrosis Factor-alpha

A diet containing different amounts of vitamin E (alpha-tocopherol; 0.5 mg, 5 mg, 10 mg or 50 mg per 100 g diet) was supplemented to BALB/c mice for 6 weeks. These mice were subcutaneously immunized twice with ovalbumin (OVA). A passive cutaneous anaphylaxis (PCA) analysis demonstrated that the mice fed on the diet containing 5 mg of vitamin E produced the highest level of the OVA-specific immunoglobulin E (IgE) antibody. A lower level of serum IgE was found in the mice supplemented with 0.5 mg, 10 mg and 50 mg of vitamin E. A sandwich ELISA analysis showed that the pattern of the total IgE antibody level among these four groups was the same as that of the allergen-specific IgE. In a separate experiment, 5 mg of vitamin E and/or 50 mg of beta-carotene was supplemented to the basal diet containing vitamin E as alpha-tocopherol acetate (5 mg) in order to evaluate the effect of their combination on OVA-specific and total IgE production in the mice. The supplementation with beta-carotene alone had no effect on OVA-specific or total IgE production. In contrast, supplementation with vitamin E plus beta-carotene effectively suppressed both the antigen-specific and total IgE antibodies. The serum vitamin E and beta-carotene levels were increased by supplementation with the respective compounds. These results strongly suggest that the combination of dietary vitamin E and beta-carotene suppressed IgE production and would therefore help to prevent the type-I allergic reaction.

Topics: Animals; Antibodies; Antibody Formation; beta Carotene; Dietary Supplements; Drug Therapy, Combination; Female; Hypersensitivity; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Rats, Wistar; Vitamin E

Chronic cellular inflammation and airway wall remodelling with subepithelial fibrosis and airway smooth muscle (ASM) cell hyperplasia are features of chronic asthma. Jun N-terminal kinase (JNK) may be implicated in these processes by regulating the transcriptional activity of activator protein (AP)-1. We examined the effects of an inhibitor of JNK, SP600125 (anthra [1,9-cd] pyrazole-6 (2 H)-one), in a model of chronic allergic inflammation in the rat. Rats sensitised to ovalbumin (OA) were exposed to OA-aerosol every third day on six occasions and were treated with SP600125 (30 mg kg-1 b.i.d; 360 mg in total) for 12 days, starting after the second through to the sixth OA exposure. We measured eosinophilic and T-cell inflammation in the airways, proliferation of ASM cells and epithelial cells by incorporation of bromodeoxyuridine (BrdU), and bronchial responsiveness to acetylcholine. SP600125 significantly reduced the number of eosinophils (P<0.05) and lymphocytes (P<0.05) in bronchoalveolar lavage fluid, suppressed eosinophilic (P<0.05) and CD2+ T-cell (P<0.05) infiltration within the bronchial submucosa, and the increased DNA incorporation in ASM (P<0.05) and epithelial cell incorporation (P<0.05). SP600125 did not alter bronchial hyper-responsiveness observed after chronic allergen exposure. Pathways regulated by JNK positively regulate ASM cell proliferation and allergic cellular inflammation following chronic allergen exposure.

Topics: Actins; Animals; Anthracenes; Bronchi; Bronchoalveolar Lavage Fluid; Cell Count; Cell Division; Chronic Disease; Eosinophils; Hypersensitivity; Immunohistochemistry; Inflammation; JNK Mitogen-Activated Protein Kinases; Male; Mitogen-Activated Protein Kinases; Myocytes, Smooth Muscle; Ovalbumin; Rats; Rats, Inbred BN; Respiratory Mucosa

Indoor formaldehyde (FA) might worsen allergies and be an underlying factor for the increasing incidence and severity of asthma; the exact mechanism, however, remains unclear.. The present study examined the effects of repeated exposure to FA on methacholine- and antigen-induced bronchoconstriction in guinea-pigs in vivo.. First, non-sensitized guinea-pigs were transnasally treated with 0.1 or 1.0% FA or saline three times a week for 6 weeks, and increasing concentrations of methacholine (50, 100, and 200 microg/mL) were inhaled at 5-min intervals. Second, guinea-pigs pre-treated with transnasal administration of FA or saline using the same protocol were passively sensitized with anti-ovalbumin (OA) serum 7 days before antigen challenge. Third, guinea-pigs were actively sensitized with OA and pre-treated with transnasal administration of FA or saline using the same protocol. The lateral pressure of the tracheal tube (Pao) was measured under anesthesia and artificial ventilation.. The antigen-induced increase in Pao in actively sensitized guinea-pigs was significantly potentiated by FA exposure in a dose-dependent manner. The dose-response curve of the methacholine-induced increase in Pao in non-sensitized guinea-pigs or of the antigen-induced increase in Pao in passively sensitized guinea-pigs was not altered by FA exposure. Transnasal administration of FA significantly increased the serum anti-OA homocytotropic antibody titre (IgG) as measured by the passive cutaneous anaphylaxis reaction in actively sensitized guinea-pigs.. The results suggest that repeated exposure to FA worsens allergic bronchoconstriction through enhancing antigen sensitization.

Topics: Allergens; Animals; Antibodies; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Fixatives; Formaldehyde; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Methacholine Chloride; Ovalbumin; Passive Cutaneous Anaphylaxis

CD4+CD25+ regulatory T cells (Treg) are instrumental in the maintenance of immunological tolerance. One critical question is whether Treg can only be generated in the thymus or can differentiate from peripheral CD4+CD25- naive T cells. In this paper, we present novel evidence that conversion of naive peripheral CD4+CD25- T cells into anergic/suppressor cells that are CD25+, CD45RB-/low and intracellular CTLA-4+ can be achieved through costimulation with T cell receptors (TCRs) and transforming growth factor beta (TGF-beta). Although transcription factor Foxp3 has been shown recently to be associated with the development of Treg, the physiological inducers for Foxp3 gene expression remain a mystery. TGF-beta induced Foxp3 gene expression in TCR-challenged CD4+CD25- naive T cells, which mediated their transition toward a regulatory T cell phenotype with potent immunosuppressive potential. These converted anergic/suppressor cells are not only unresponsive to TCR stimulation and produce neither T helper cell 1 nor T helper cell 2 cytokines but they also express TGF-beta and inhibit normal T cell proliferation in vitro. More importantly, in an ovalbumin peptide TCR transgenic adoptive transfer model, TGF-beta-converted transgenic CD4+CD25+ suppressor cells proliferated in response to immunization and inhibited antigen-specific naive CD4+ T cell expansion in vivo. Finally, in a murine asthma model, coadministration of these TGF-beta-induced suppressor T cells prevented house dust mite-induced allergic pathogenesis in lungs.

Topics: Animals; CD4 Antigens; DNA-Binding Proteins; Forkhead Transcription Factors; Gene Expression Regulation; Hypersensitivity; Immune Tolerance; Immunophenotyping; Interleukin-2; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mites; Ovalbumin; Receptors, Antigen, T-Cell; Receptors, Interleukin-2; T-Lymphocytes; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

This report examines the effect of heat-killed Mycobacterium vaccae in a mouse model of allergic pulmonary inflammation. The s.c. administration of M. vaccae 3 wk before the immunization significantly reduced Ag-induced airway hyperreactivity and the increase in the numbers of eosinophils observed in the bronchoalveolar lavage fluid, blood, and bone marrow, even though no detectable changes in either cytokine (IL-4, IL-13, IL-5, and IFN-gamma) or total IgE levels were observed. Furthermore, transfer of splenocytes from OVA-immunized and M. vaccae-treated mice into recipient, OVA-immunized mice significantly reduced the allergen-induced eosinophilia by an IFN-gamma-independent mechanism, clearly indicating that the mechanism by which M. vaccae induces its inhibitory effect is not due to a redirection from a predominantly Th2 to a Th1-dominated immune response. The protective effect of M. vaccae on the allergen-induced eosinophilia lasted for at least 12 wk after its administration, and the treatment was also effective in presensitized mice. Moreover, the allergen specificity of the inhibitory effect could be demonstrated using a double-immunization protocol, where M. vaccae treatment before OVA immunization had no effect on the eosinophilic inflammation induced by later immunization and challenge with cockroach extract Ag. Taken together, these results clearly demonstrate that M. vaccae is effective in blocking allergic inflammation by a mechanism independent of IFN-gamma, induces long term and Ag-specific protection, and therefore has both prophylactic and therapeutic potential for the treatment of allergic diseases.

Topics: Adoptive Transfer; Animals; Antigens; Bacterial Vaccines; Bronchial Hyperreactivity; Chemokines; Cytokines; Eosinophilia; Eosinophils; Female; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Inflammation; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; Vaccines, Inactivated

Rats are commonly used in anaphylaxis models, mainly in intestinal anaphylaxis. Hypersensitivity mechanisms are complex and they are not clearly defined. Ovalbumin (OVA) is commonly used for studies on the hypersensitivity mechanism. However, the potential pro-inflammatory mediators induced by this antigen in the model of paw oedema in immunized rats are still not completely understood. This work examines the pharmacological modulation of several mediators involved in rat hind paw immune oedema induced by OVA. Wistar rats were previously immunized (14-18 days) with OVA (30 microg, intraperitoneally) or sham-sensitized with aluminum hydroxide (control). The paw volumes were measured before the antigenic stimuli and 1, 2, 3 and 4 h after the intraplantar injection of OVA (10 microg/paw). Subcutaneous injection of dexamethasone, diphenhydramine, cyproheptadine, chlorpromazine or methysergide significantly inhibited (p < 0.05) the allergic paw oedema. The dual inhibitor of cyclooxygenase and lipoxygenase (NDGA), the cyclooxygenase inhibitor (indomethacin), the lipoxygenase inhibitor (MK-886), the PAF antagonist (WEB 2086), the mast cell stabilizer (ketotifen), and the anti-histamine (meclizine) did not inhibit the immune oedema. In addition, thalidomide and pentoxifylline (anti-tumour necrosis factor drugs) were ineffective against OVA-induced oedema. The fact that indomethacin, MK-886, NDGA and WEB 2086 are unable to inhibit this allergic oedema indicates that the dexamethasone action seems not to be via phospholipase A2, but possibly due to the synthesis and/or the inhibitory activity of cytokines. The paw oedema inhibition by diphenhydramine, but not by meclizine, may suggest a different mechanism, which is independent of the effect of histamine. These data indicate that allergic oedema is more sensitive to anti-serotonin drugs, mainly anti-5-HT2, suggesting that the principal mediator of this inflammatory response is serotonin.

Topics: Animals; Anti-Allergic Agents; Azepines; Chlorpromazine; Dopamine Antagonists; Dose-Response Relationship, Immunologic; Edema; Enzyme Inhibitors; Foot; Histamine H1 Antagonists; Hypersensitivity; Immunization; Immunosuppressive Agents; Ketotifen; Ovalbumin; Platelet Aggregation Inhibitors; Rats; Rats, Wistar; Triazoles

Interferon (IFN)-gamma reduces airway responses after allergen challenge in mice. The mechanisms of this effect are not clear. These studies investigate whether IFN-gamma can reverse prolonged airway responses after allergen challenge in IFN-gamma-deficient (IFN-gammaKO) mice. Sensitized mice (IFN-gammaKO and wild-type [WT]) were challenged with ovalbumin. Airway responsiveness, eosinophils in bronchoalveolar lavage fluid, and lung lymphocyte subsets (CD4(+) and CD8(+)) were measured 24 hours and 8 weeks after challenge. In further experiments, we treated IFN-gammaKO mice with recombinant IFN-gamma starting 4 weeks after the challenge for 1 week or 4 weeks. Airway responsiveness, bronchoalveolar lavage eosinophils, and lung CD4(+) cells were increased 8 weeks after challenge in IFN-gammaKO but not WT mice. IFN-gamma treatment returned lung CD4(+) cell numbers to values obtained in unchallenged mice. One week of IFN-gamma treatment also returned airway responsiveness to baseline levels; however, 4-week treatment with IFN-gamma failed to decrease airway responsiveness below levels observed in untreated animals. This suggests that IFN-gamma plays an essential role in reversing allergen-induced airway inflammation and hyperresponsiveness and that it may have dual actions on the latter. Observations that IFN-gamma reverses airway responses, even when administered after challenge, suggests that IFN-gamma treatment could control allergic disease, including asthma.

Topics: Airway Obstruction; Animals; Animals, Wild; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; CD4-CD8 Ratio; Disease Models, Animal; Drug Evaluation, Preclinical; Eosinophils; Female; Humans; Hypersensitivity; Inflammation; Interferon-gamma; Mice; Mice, Inbred BALB C; Ovalbumin

The effects of chronic unpredictable stress on airway leukocyte infiltration and plasma extravasation in female rats have been investigated. The chronic stress lasted for 14 days and consisted of transitory and variable changes in the living conditions of the animals. Concomitant to the stress procedure, the animals were sensitized (Day 0) and challenged with ovalbumin (OVA; 200 microg) at Day 14. Bronchoalveolar lavage (BAL) was performed 48 h after intratracheal challenge with OVA (0.4 ml of a 0.25% solution). The increase in plasma extravasation was assessed by the rat paw oedema induced by OVA (0.1 mg/paw) or the mast cell degranulator compound 48/80 (5 microg/paw). A significant increase (P < .05) in the total leukocyte influxes into the airways was observed in the stressed (sensitized) group compared to nonstressed (sensitized) animals, and this was associated with a marked recruitment of eosinophils and mononuclear cells in the BAL fluid. In OVA-sensitized rats, intraplantar injection of OVA induced a marked paw oedema that was significantly higher in stressed compared to nonstressed groups. In contrast to OVA, the compound 48/80 (5 microg/paw)-induced oedema did not significantly differ between nonstressed and stressed groups. Our results indicate that chronic unpredictable stress exacerbates the vascular and cellular inflammatory responses.

Topics: Animals; Bronchoalveolar Lavage Fluid; Female; Hypersensitivity; Leukocyte Count; Ovalbumin; p-Methoxy-N-methylphenethylamine; Rats; Rats, Wistar; Stress, Physiological

The immune status and allergen exposure of the mother may influence the immune response in the offspring after birth. This relationship may be important both for allergen avoidance strategies and, alternatively, for allergy prophylaxis by allergen exposure of the mother.. The aim of the present study was to investigate the effect of allergen immunization of the mother during pregnancy and postpartum, in relation to the allergy-related immune response (IgE) and the non-allergy-related (IgG2a) response in the offspring.. Pregnant NIH/OlaHsd females were immunized three times during pregnancy and one time postpartum with ovalbumin and the adjuvant Al(OH)3, and the offspring's ovalbumin-specific IgE, IgG1 and IgG2a responses were measured after challenge with the same allergen as young adults. Ovalbumin-specific IgE, IgG1 and IgG2a responses were also analysed in offspring of NIH/OlaHsd females immunized once at different times during pregnancy: about 3 days into pregnancy, mid-pregnancy (10 days into pregnancy) and about 4 days before giving birth (17 days into pregnancy).. Allergen immunization of mother during pregnancy and postpartum significantly reduced the IgE response in the progenies, whereas the IgG2a response to the same allergen was increased. Allergen immunization of the mother 3 days into pregnancy resulted in a significantly lower IgE response in offspring compared with the response in offspring of non-immunized mothers and in offspring of mothers immunized 17 days into pregnancy.. Maternal allergen immunization might favour selection for an allergen-specific Th1-dependent antibody response in the offspring. Our results indicate that IgE suppression is stronger after maternal allergen exposure during early pregnancy than after exposure in late pregnancy.

Topics: Adjuvants, Immunologic; Allergens; Animals; Female; Gestational Age; Hypersensitivity; Immunity, Maternally-Acquired; Immunization; Immunization Schedule; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred Strains; Models, Animal; Ovalbumin; Pregnancy

This study was designed to evaluate the value and applicability of tidal breathing pattern analysis to assess bronchoconstriction in conscious rats. Using noninvasive, head-out body plethysmography and the decrease in tidal midexpiratory flow (EF(50)), we measured airway responsiveness (AR) to inhaled acetylcholine and allergen in conscious Brown-Norway rats, followed by invasive determination of pulmonary conductance (GL) and EF(50) in anesthetized rats. Dose-response studies to acetylcholine showed that noninvasively recorded EF(50) closely reflected the dose-dependent decreases observed with the invasive monitoring of simultaneously measured GL and EF(50). After sensitization and intratracheal boost to ovalbumin or saline, rats were assessed for early and late AR to aerosolized ovalbumin. Ovalbumin aerosol challenge resulted in early and late AR in allergen-sensitized rats, whereas controls were unresponsive. The allergen-specific AR, as measured noninvasively by EF(50), was similar in degree compared with invasively recorded EF(50) and GL and was associated with enhanced IgE and airway inflammation. We conclude that EF(50) is a noninvasive and physiologically valid index of bronchoconstriction in a rat model of asthma.

Topics: Acetylcholine; Administration, Inhalation; Allergens; Animals; Bronchoconstriction; Carbon Dioxide; Hypersensitivity; Male; Maximal Midexpiratory Flow Rate; Ovalbumin; Plethysmography; Pulmonary Ventilation; Rats; Rats, Inbred BN; Respiration; Respiratory System

The effects of the dietary oligosaccharide raffinose on immune responses, with special reference to its anti-allergic functions, were examined in vivo. First, feeding a diet supplemented with 50 g raffinose/kg to BALB/c mice significantly (P<0.05) increased interleukin (IL) 12 secretion from isolated Peyer's patch (PP) cells in vitro compared with feeding control diet. When isolated PP cells were used as antigen-presenting cells (APC) for CD4+ T-splenocytes isolated from ovalbumin (OVA)-specific T-cell receptor transgenic (Tg) mice in the presence of OVA as antigen, significantly (P<0.05) higher levels of interferon-gamma were observed in the cultures using APC from raffinose-fed mice than those cultures using APC from control mice. Second, the diet containing 50 g raffinose/kg or control diet was fed to OVA Tg mice, and subsequently, OVA was added to each diet to prime T cells in vivo. CD4+ T-cells from the mesenteric lymph nodes of the raffinose-fed mice secreted significantly (P<0.05) higher levels of IL-2 and significantly (P<0.05) lower levels of IL-4 following in vitro antigenic stimulation compared with those of the control mice. These present results suggest that feeding raffinose may suppress differentiation of naïve T-helper (Th) cells into Th2 cells in the mesenteric lymphoid nodes. Last, feeding raffinose suppressed rises of serum immunoglobulin E levels in the Tg mice treated with long-term ingestion of OVA. In conclusion, it is suggested that dietary raffinose suppresses serum immunoglobulin E response through suppression of Th2-type immune response against oral antigen in the lymphoid organs located in or near the intestine.

Topics: Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Dose-Response Relationship, Immunologic; Hypersensitivity; Immunoglobulin E; Interleukin-12; Interleukin-2; Interleukin-4; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Transgenic; Models, Animal; Ovalbumin; Peyer's Patches; Raffinose; Receptors, Antigen, T-Cell; Th2 Cells

The role of nitric oxide (NO) in allergic inflammation and bronchial hyperresponsiveness is unclear. We studied a selective prodrug nitric oxide synthase (NOS)-2 inhibitor, L-N(6)-(1-iminoethyl)lysine 5-tetrazole amide (SC-51). In ovalbumin-sensitized and challenged rats, exhaled NO levels increased by 3 h following challenge (3.73 +/- 0.74 ppb; P < 0.05), peaking at 9 h (11.0 +/- 2.75; P < 0.01) compared to saline controls (1.87 +/- 0.26; P < 0.05 and 2.81 +/- 0.18; P < 0.01). Immunoreactive lung NOS2 expression was increased in ovalbumin-challenged rats compared with ovalbumin-sensitized, saline-challenged rats at 8 h post-challenge. SC-51 (10 mg/kg; p.o.) inhibited allergen-induced increase in exhaled NO levels to 1.3 +/- 0.17 ppb. SC-51 inhibited bronchial hyperresponsiveness in ovalbumin-sensitized and challenged rats (P < 0.05). In sensitized non-exposed rats, SC-51 increased bronchial responsiveness (P < 0.05). SC-51 reduced the allergen-induced increase in bronchoalveolar lavage neutrophils, but caused a nonsignificant reduction in bronchial mucosal eosinophil numbers. NO generated through NOS2 contributes to allergen-induced bronchial hyperresponsiveness but not to bronchial eosinophilia, indicating that these are independently expressed.

Topics: Acetylcholine; Airway Resistance; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Enzyme Inhibitors; Eosinophils; Homoarginine; Hypersensitivity; Immunohistochemistry; Inflammation; Isoenzymes; Lung; Male; Nitric Oxide; Nitric Oxide Synthase; Ovalbumin; Rats; Rats, Inbred BN

Exposure to environmental tobacco smoke (ETS) has been shown to increase allergic sensitization and reactivity and there has been some suggestion that the influence of ETS on the allergic response is dissimilar in males and females. It is to be determined whether gender differences exist in the IgE response to ovalbumin (OVA) sensitization following ETS exposure from the neonatal period through adulthood. To address this thesis, we examined gender differences in OVA sensitization of BALB/c mice housed from birth through adulthood under smoking and nonsmoking conditions. At 6 weeks of age (day 0) all mice were injected i.p. with OVA in aluminum hydroxide adjuvant followed by three 20 min exposures to 1% aerosolized OVA between day 14 and 80. There were significantly (p < 0.05) more total and OVA specific IgE and IgG1 in the serum of females compared to males. Moreover, these sex responses, along with eosinophilia, were further enhanced in mice exposed to ETS. There were also significantly more IgE positive cells in the lungs of female, but not male, mice exposed to ETS compared with ambient air (p < 0.05). There was also an elevation of Th2 cytokines (IL4, IL5, IL10, and IL13) after re-stimulation of lung homogenates following ETS exposure. These data demonstrate that female animals are significantly more susceptible than males to the influence of ETS on the allergic response.

Topics: Animals; Animals, Newborn; Cytokines; Eosinophils; Female; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Leukocyte Count; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Sex Characteristics; Tobacco Smoke Pollution

T cell activation and cytokine secretion are important mediators of inflammation in allergic asthma. The costimulatory pathway CD28/CD80/CD86 has been shown to play an important role in T cell activation in allergic asthma, but less is known about the effect of other costimulatory molecules in allergy. The costimulatory molecule OX40 ligand (OX40L), a member of the tumor necrosis factor superfamily, has been shown to be important in T cell priming and cytokine production. We investigated the role of OX40L in a murine model of allergic inflammation using OX40L(-/-) mice. In this model, following OVA sensitization and challenge, mice develop features of allergic inflammation including elevated levels of total serum IgE, pulmonary eosinophils, cytokines, and pulmonary inflammation. In the absence of OX40L, total serum IgE, pulmonary eosinophils, cytokines, and pulmonary inflammation were all significantly reduced compared to wild-type controls. Levels of eotaxin mRNA, an eosinophil-specific chemoattractant, were also markedly reduced, paralleling the significant reduction in pulmonary eosinophils. Levels of allergen-induced Th1 as well as Th2 cytokines were also significantly reduced. Together, the data support a critical role for OX40L signals in allergic responses.

Topics: Animals; Antigens, CD; Asthma; B7-1 Antigen; B7-2 Antigen; CD28 Antigens; Cytokines; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Ovalbumin; OX40 Ligand; Th1 Cells; Th2 Cells; Tumor Necrosis Factors

Both innate and specific defenses of the preterm infant are even less developed than those of term infants, and the immune systems of preterm infants might be skewed differently at birth. Their immune responses to food antigens started early in life might therefore differ from those of term infants.. We sought to compare antibody levels to cow's milk, ovalbumin, and gliadin at age 10 years in children who had been born either preterm or at term.. IgG and IgA isotype antibodies to whole cow's milk, beta-lactoglobulin, alpha-casein, and ovalbumin, as well as IgG antibody levels to gliadin and to tetanus and diphtheria toxoids, were measured for a group of 62 children born preterm and 61 control subjects born at term. These children were studied at the same time for atopy.. Children born preterm had markedly lower levels of antibodies to cow's milk and to its protein fractions (P <.0001 for IgA and IgG antibodies to cow's milk and alpha-casein and IgG beta-lactoglobulin antibodies). IgG gliadin antibodies were also significantly lower in the preterm group (P =.03), although the difference was not significant for IgG ovalbumin antibodies. In the preterm group both those born before gestational week 30 and those given cow's milk-based formula early (before day 50) had the lowest levels of cow's milk antibodies. In the preterm group atopy was associated with low levels of IgG cow's milk antibodies but with high levels of IgG ovalbumin antibodies.. Early introduction of food antigens into the immature gastrointestinal tract of preterm infants might result in tolerance. The presence of less atopy in these children might also be a result of tolerance development.

Topics: Animals; Antibodies; Child; Female; Gliadin; Humans; Hypersensitivity; Immunoglobulin A; Immunoglobulin G; Infant Food; Infant, Newborn; Infant, Premature; Male; Milk; Milk Proteins; Ovalbumin; Reference Values

Bronchial inflammation in allergic asthma is associated with active exudation from the bronchial tree into the interstitial space of both mucosa and submucosa. The aim of this study was to evaluate epithelial and endothelial permeability as well as alveolar fluid movement in a model of chronic allergic inflammation in Brown-Norway rats sensitized and challenged with ovalbumin (OA). Control groups were challenged with saline solution (C), and rats were immunized by OA but not challenged (Se). Lung sections showed a marked inflammatory infiltrate associated with perivascular and peribronchiolar edema in OA. To measure alveolar liquid clearance, a 5% bovine albumin solution with 1 microCi of (125)I-labeled human albumin was instilled into the air spaces. Alveolar-capillary barrier permeability was evaluated by intravascular injection of 1 microCi of (131)I-labeled albumin. Endothelial permeability was significantly increased in OA, from 0.08 +/- 0.01 in the C group to 0.19 +/- 0.03 in OA group (P < 0.05). Final-to-initial protein ratio was also statistically higher in OA (1.6 +/- 0.05) compared with C (1.38 +/- 0.03, P = 0.01) and Se groups (1.42 +/- 0.03, P = 0.04). Administration of anti-tumor necrosis factor-alpha antibodies within the instillate significantly decreased this ratio (1.32 +/- 0.08, P = 0.003 vs. OA). To conclude, we demonstrated a tumor necrosis factor-alpha-dependent increase in alveolar fluid movement in a model of severe bronchial allergic inflammation associated with endothelial and epithelial leakage.

Topics: Albumins; Animals; Blood-Air Barrier; Body Fluids; Bronchitis; Capillaries; Capillary Permeability; Cattle; Chronic Disease; Humans; Hypersensitivity; Immunization; Immunoglobulin E; Lung; Male; Ovalbumin; Pulmonary Alveoli; Rats; Rats, Inbred BN; Tumor Necrosis Factor-alpha

We have previously shown that lipopolysaccharide (LPS) exposure in sensitised animals 18 h after ovalbumin (OVA) challenge inhibits OVA-induced airway hyper-responsiveness (AHR). In the present study, we investigated the effect of LPS on OVA-induced acute and late-phase allergic responses in sensitised rats when challenged with OVA.. Rats were sensitised with OVA and 11 days later challenged with 1% OVA in the presence or absence of LPS (0.5-50 microg/ml) given in the same nebulizer. Acute responses to OVA were measured each minute for 30 min after challenge. In a separate group of animals, late-phase responses to OVA were determined at 24 h. At the end of each study, Evans blue dye was injected and animals sacrificed 30 min later. Bronchoalveolar lavage was obtained to monitor inflammatory cell migration and microvascular leakage.. OVA challenge in sensitised animals produced an acute response with changes in lung mechanics peaking 10.0 +/- 0.9 min after OVA and returning to baseline within 30 min. This was followed 24 h later by increased responses to methacholine chloride (MCh), inflammatory cell influx and increased Evans blue leakage into the lungs. Presence of 5 or 50 microg/ml LPS in the nebulizer during OVA challenge altered the kinetics of the acute-phase response, with an immediate decrease in lung function (time to peak decreased from 10.3 +/- 1.2 to 1.8 +/- 0.2 and 2.2 +/- 0.3 min, respectively: p < 0.001, n = 6) and a dose-dependent attenuation of late-phase AHR, cellular influx (n = 5, p < 0.001) and Evans blue leakage (n = 5, p < 0.001) at 24 h.. In summary, co-administration of OVA with LPS modifies both the acute and late-phase responses to the allergen, inducing an earlier acute change in lung function and a dose-dependent inhibition of late-phase responses to the allergen.

Topics: Acute-Phase Reaction; Animals; Bronchoalveolar Lavage Fluid; Coloring Agents; Enzyme-Linked Immunosorbent Assay; Evans Blue; Hypersensitivity; Immunoglobulin E; Lipopolysaccharides; Male; Ovalbumin; Rats; Respiratory Function Tests; Statistics, Nonparametric

Toxic influence of high oxygen concentration on pulmonary function and structures has been known for many years. However, the influence of high oxygen concentration breathing on defensive respiratory reflexes is still not clear. In our previous experiments, we found an inhibitory effect of 100 % oxygen breathing on cough reflex intensity in healthy guinea pigs. The present study was designed to detect the effects of hyperoxia on cough reflex in guinea pigs with allergic airway inflammation. In the first phase of our experiment, the animals were sensitized with ovalbumin. Thirty-two sensitized animals were used in two separate experiments according to oxygen concentration breathing: 100 % or 50 % oxygen for 60 h continuously. In each experiment, one group of animals was exposed to hyperoxia, another to ambient air. The cough reflex was induced both by aerosol of citric acid before sensitization, then in sensitized animals at 24 h and 60 h of exposition to oxygen/air in awake animals, and by mechanical stimulation of airway mucosa in anesthetized animals just after the end of the experiment. In contrast to 50 % oxygen, 100 % oxygen breathing leads to significant decrease in chemically induced cough in guinea pigs with allergic inflammation. No significant changes were present in cough induced by mechanical stimulation of airways.

Topics: Animals; Bronchial Hyperreactivity; Citric Acid; Cough; Female; Guinea Pigs; Hyperoxia; Hypersensitivity; Ovalbumin; Oxygen; Reflex; Respiratory Mechanics

Lactoferrin (LF) is an iron-binding glycoprotein present in the cytoplasmic granules of neutrophils and in external secretions of mammals. Although the biological role of human and bovine lactoferrin has been extensively studied, there is still uncertainty as to the nature and function of lactoferrin receptors. We recently determined that methyl-alpha-D-mannopyranoside given intraperitoneally (i.p.) could suppress the adjuvant activity of LF in the generation of delayed-type hypersensitivity (DTH) to ovalbumin (OVA). We concluded that the lactoferrin effects in DTH are mediated by carbohydrate-recognizing receptors like the mannose receptor (MR). This study indicates that subcutaneous (s.c.) administration of very small doses of the Man-bovine serum albumin (Man-BSA) complex, together with a sensitizing dose of the antigen, gives the same effects as i.p. administration of methyl-alpha-D-mannopyranoside. The latter is also a blocker of MR, although of a much lower affinity to the receptor than Man-BSA. The blocking of the adjuvant effect of LF by the Man-BSA complex (when given together with the sensitising dose of antigen) suggests that the function of antigen-presenting cells in the skin (presumably immature dendritic cells expressing MR) is inhibited. The results of our study indicate that a receptor with an affinity for mannose is essential for the mediation of adjuvant lactoferrin function in the generation of DTH.

Topics: Adjuvants, Immunologic; Animals; Female; Hypersensitivity; Lactoferrin; Male; Mannose; Mice; Mice, Inbred CBA; Ovalbumin; Serum Albumin, Bovine

During the last decades, the prevalence of the allergic airway diseases, asthma and rhinitis, has increased world-wide. Introduction of environmental chemicals with adjuvant effect may play a role in this increase. In the present study, the adjuvant effects of di-n-butyl-, di-n-octyl-, di-iso-nonyl- and di-iso-decyl phthalate are studied in a screening model. Ovalbumin, used as the model antigen, was injected subcutaneously in the neck region of BALB/cJ mice with the selected phthalate in concentrations from 2-2000 microg/ml. Additionally, the mice were boosted once or twice with ovalbumin alone. Immunization with ovalbumin alone, the ovalbumin control group, served as the baseline for antibody production, whereas aluminium hydroxide served as the positive control. The levels of ovalbumin-specific IgE, IgG1 and IgG2a antibodies in sera were determined. Adjuvant effect was accepted to be present if a statistical increase in antibody production occurred in a test group as compared to an ovalbumin control group together with the fulfillment of dose-response relationships. Adjuvant effect varied strongly between the phthalates investigated. Phthalates with 8 or 9 carbon atoms in the alkyl side chains were the stronger adjuvants whereas phthalates with shorter or longer alkyl side chains possessed less adjuvant activity. Adjuvant effects were apparent either from the IgE or the IgG1 response or both, whereas no effect was seen on the IgG2a response. Additional studies with airborne exposure are required to establish whether the hazards also result in a significant risk for the development of allergy in man.

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Dibutyl Phthalate; Dose-Response Relationship, Drug; Environmental Pollutants; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Injections, Subcutaneous; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalic Acids; Plasticizers; Structure-Activity Relationship

We recently reported that allergic lung inflammation in guinea pigs became steroid resistant in the presence of latent adenoviral infection.. We sought to investigate the molecular mechanisms that underlie steroid resistance in adenoviral infection.. Guinea pigs with a latent adenoviral infection were sensitized and challenged with ovalbumin (OVA) and given daily injections of budesonide (20 mg/kg administered intraperitoneally). Sham-infected animals received either saline challenge without budesonide injection or OVA challenge with or without budesonide. The inflammatory response in the lung was measured by means of quantitative histology. Eotaxin, monocyte chemoattractant protein 1 (MCP-1), and RANTES expression in the lung were analyzed by means of Northern blotting, and the binding activity of activator protein 1 (AP-1) and nuclear factor kappaB in nuclear extracts from the lung was analyzed with electrophoretic mobility shift assays.. OVA challenge increased eosinophil infiltration and eotaxin and MCP-1 mRNA expression in the lungs, and glucocorticoids reduced these increases in the sham-infected, but not the adenovirus-infected, animals. Changes in binding activity of AP-1, but not nuclear factor kappaB, paralleled changes in eotaxin and MCP-1 mRNA.. We conclude that latent adenoviral infection inhibits the anti-inflammatory effects of glucocorticoids on allergen-induced eotaxin and MCP-1 expression through AP-1, leading to steroid-resistant allergic lung inflammation.

Topics: Adenoviridae Infections; Animals; Chemokines; DNA; Drug Resistance; Female; Glucocorticoids; Guinea Pigs; Hypersensitivity; NF-kappa B; Ovalbumin; Pneumonia; RNA, Messenger; Transcription Factor AP-1

Although surfactant apoproteins are known to be mediators of innate responses, their relationship to adaptive responses has not been examined extensively. We investigated possible links between surfactant apoproteins and responses to allergens by studying alterations in surfactant apoproteins A, B, and D in a murine model of allergic pulmonary inflammation. Three murine strains (BALB/c, C57BL/6, and 129J) demonstrated increased immunostaining of surfactant apoproteins A and D in nonciliated epithelial cells of noncartilaginous airways after aerosolized challenge. In contrast, surfactant apoprotein B immunostaining was unchanged. Immunoblotting demonstrated increased surfactant A in bronchoalveolar lavage fluid after allergen sensitization and challenge. Surfactant apoprotein A and D induction required T and/or B lymphocyte responses to allergen, since the induction was absent in recombinase-activating gene-deficient mice, which lack functional lymphocytes. We conclude that increased immunoreactivity of two collectins, surfactant apoproteins A and D, occurs within the response to allergen. Our findings support a model in which surfactant apoproteins A and D are important to both innate immunity and adaptive immune responses to allergens.

Topics: Adaptation, Physiological; Animals; Antibody Formation; Apoproteins; B-Lymphocytes; Bronchoalveolar Lavage Fluid; Carrier Proteins; Collectins; Genes, RAG-1; Glycoproteins; Hypersensitivity; Immunization; Lung; Mice; Mice, Inbred Strains; Mice, Knockout; Ovalbumin; Proteolipids; Pulmonary Surfactant-Associated Protein D; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; T-Lymphocytes

Patients with allergic asthma present clinically with chronic or intermittent disease caused by either persistent or periodic allergen exposure. We sought to generate clinically relevant disease in mice, which would reflect the relapsing, remitting, and constant nature of this syndrome. We generated and compared acute onset, remission, relapse, and overt phases of the disease and found that acute disease was characterized by airway hyperreactivity, eosinophilic lung inflammation, excessive mucus production, and antigen-specific antibody and was rapidly followed by a remission. Mice rechallenged with aerosol antigen during the remission or treated with repeated aerosol challenges developed relapse and overt disease, respectively. Recurrent antigen exposure induced a progressive increase in bronchoalveolar lavage fluid immunoglobulin, mucus production, and a change in inflammatory infiltrates indicating a transition from acute to chronic inflammation. These data demonstrate distinct phases of disease representing a clinical spectrum of experimental allergic asthma and may have important implications for new treatment strategies.

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Recurrence

Th2 lymphocyte responses are associated with inflammation and disease during allergic responses. Exposure to particular environmental factors during the expression of allergy could result in more pronounced Th2-like immune responses and more severe disease. One factor might be a respiratory virus infection.. The aim of our study was to investigate the influence of respiratory syncytial virus (RSV) infection on the expression of ovalbumin (OVA)-induced allergy in BALB/c mice.. We determined OVA-specific IgE in serum, cytokine profiles and histopathological lesions in lungs of OVA-allergic mice after RSV infection.. OVA sensitization and challenge induced OVA-specific IgE in serum, Th2 cytokine mRNA expression, and mononuclear and eosinophilic inflammation in the lungs. RSV inoculation during the challenge period enhanced OVA-induced IL-4 and IL-5 mRNA expression in lung tissue. RSV further enhanced the OVA-induced hypertrophy of mucous cells and eosinophilic infiltration in lung tissue. Surprisingly, RSV infection decreased Th2 cytokine secretion and eosinophilic influx in bronchoalveolar lavage of OVA-allergic mice. Because inactivated RSV did not influence these responses, replication of RSV appeared essential for the modification of OVA-induced Th2 cytokine expression. RSV did not change OVA-specific IgE levels in serum. Furthermore, the RSV-induced IL-12 mRNA expression in lung tissue of OVA-allergic mice was diminished, but IFN-gamma mRNA expression was not affected.. RSV infection enhanced particular OVA-induced Th2 cytokine mRNA responses and pulmonary lesions in allergic mice and thus aggravated allergic respiratory disease.

Topics: Animals; Antibody Specificity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human; RNA, Messenger; Severity of Illness Index; Time Factors; Ultraviolet Rays

To better understand asthma pathogenesis, we characterized airway responsiveness, lung inflammation, and mediator production of alveolar macrophages (AM) after allergen sensitization and challenge in two strains of rats showing different susceptibilities in developing airway allergic reactions. Airway responsiveness to acethylcholine was measured 24 h after ovalbumin (OVA) challenge, whereas bronchoalveolar lavages were performed 5 min, 8 h, and 24 h after challenge. Brown Norway rats showed airway hyperresponsiveness after challenge, whereas lung resistance remained unchanged in Sprague-Dawley rats. Interestingly, Sprague-Dawley rats developed a neutrophilic inflammation, whereas both neutrophils and eosinophils were increased in Brown Norway rats. AM mediator production varied with time with a lower tumor necrosis factor (TNF) and interleukin (IL)-10 release at 8 h after challenge. OVA challenge stimulated spontaneous TNF and IL-10 release by AM isolated 24 h after challenge in both strains of rats, although AM from Brown Norway rats released significantly more IL-10 and TNF. Furthermore, nitric oxide production was increased only in OVA-challenged (24 h) Brown Norway rats. Our results suggest that AM may participate to the expansion of Th2 inflammation in Brown Norway rats and that differences in AM mediator production may explain, in part, distinct allergic susceptibilities in these two strains of rats.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Progression; Disease Susceptibility; Eosinophils; Hypersensitivity; Inflammation Mediators; Interleukin-10; Lung; Macrophages, Alveolar; Neutrophils; Nitric Oxide; Ovalbumin; Pneumonia; Rats; Rats, Inbred BN; Rats, Sprague-Dawley; Species Specificity; Tumor Necrosis Factor-alpha

Inflammatory responses induced by allergen exposure cause mucous cell metaplasia (MCM) by differentiation of existing and proliferating epithelial cells into mucus-storing cells. Airway epithelia have various mechanisms that resolve these changes to form normal airway epithelia. In this report, we first investigated the state of mucous cell metaplasia and the mechanisms by which MCM is reduced despite continued exposures to allergen. After 5 days of allergen exposure, extensive MCM had developed but was reduced when allergen challenge was continued for 15 days. During this exposure period, IL-13 levels decreased and IFN-gamma levels increased in the bronchoalveolar lavage fluid. In contrast, IL-13 levels decreased but IFN-gamma was not detected at any time point during the resolution of MCM following cessation of allergen exposure. Instillation of IFN-gamma but not anti-Fas caused accelerated resolution of MCM and MCM was not resolved in Stat1-deficient mice exposed to allergen for 15 days, confirming that IFN-gamma is crucial for reducing MCM during prolonged exposures to allergen. IFN-gamma but not anti-Fas induced apoptotic cell death in proliferating normal human bronchial epithelial cells and in human bronchial epithelial cells from subjects with asthma. The apoptotic effect of IFN-gamma was caspase dependent and was inhibited by IL-13, indicating that the Th2 milieu in asthmatics may maintain MCM by preventing cell death in metaplastic mucous cells. These studies could be useful in the understanding of deficiencies leading to chronicity in airway changes and designing novel therapies to reverse MCM and airway obstruction in asthmatics.

Topics: Adult; Allergens; Animals; Apoptosis; Asthma; Bronchi; Cells, Cultured; DNA-Binding Proteins; fas Receptor; Humans; Hypersensitivity; Interferon-gamma; Interleukin-13; Kinetics; Male; Metaplasia; Mice; Mice, Inbred C57BL; Ovalbumin; Respiratory Mucosa; STAT1 Transcription Factor; Trans-Activators

The effects of interferon (IFN)-gamma induced by virus infection on eosinophil reaction in allergic airway inflammation are not yet clear. We investigated the effects of lactic dehydrogenase virus (LDV) infection, which increases IFN -gamma production with no viral infection or replication in respiratory epithelium, on allergic airway hypersensitivity. LDV infection suppressed antigen-induced eosinophil recruitment into the airway in sensitized mice. IL -5 gene expression in bronchoalveolar lavage (BAL) cells was significantly suppressed in LDV -infected mice compared with uninfected controls. The numbers of total T cells and CD 4+ T cells were significantly reduced in LDV -infected mice compared with controls. The present results suggested that the increase in production of IFN -gamma by viral infection suppresses the eosinophil reaction, and this suppressive effect may be mediated by inhibition of the recruitment of CD 4+ T cell and IL -5 production.

Topics: Allergens; Animals; Arterivirus Infections; Base Sequence; Bronchoalveolar Lavage; CD4-Positive T-Lymphocytes; Eosinophils; Gene Expression Regulation; Hypersensitivity; Interferon-gamma; Interleukin-5; Lactate dehydrogenase-elevating virus; Male; Mice; Ovalbumin; Respiratory System; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Rodent Diseases

Allergen-specific serum IgE may be insensitive as a marker for IgE-mediated reactions at the mucosal level. Five of six atopic beagle dogs developed high ovalbumin (OVA)-specific serum IgE levels after sensitization. This study aimed to show that these dogs still express allergen-specific IgE at the pulmonary and ocular mucosal levels and in the skin even when corresponding serum IgE was below the detection limit. When serum IgE levels were negative, all dogs exhibited allergic reactions at the tissue level. Specifically, they displayed positive ocular reactions after an ocular OVA challenge. After airway challenge with aerosolized OVA, five out of six animals reacted with decreased compliance and increased resistance of the lungs. Furthermore, an eosinophilia in the bronchoalveolar lavage fluid (BALF) was observed. Four weeks after the last exposure to OVA, IgE-positive BALF cells were seen in all animals. Six weeks on, all dogs still displayed positive skin reactions to OVA. This indicates that not only skin testing but also detection of ocular and pulmonary allergic tissue reactions including cell-bound IgE in BALF can serve as more sensitive and lasting surrogate markers of hypersensitivity in the allergic dog model than detection of allergen-specific serum IgE levels.

Topics: Animals; Disease Models, Animal; Dogs; Eye; Hypersensitivity; Immunoglobulin E; Lung; Ovalbumin; Skin Tests

1. We investigated the effect of MEN 11467 ((1R,2S)-2-N[1(H)indol-3-yl-carbonyl]-1-N-[N(alpha)(p-tolylacetyl)-N(alpha)(methyl)-D-3-(2-naphthyl)alanyl]diaminocyclohexane) on tachykinin-induced mucus secretion in ferret trachea in vitro and determined its effect on secretion by tracheae from allergic ferrets in response to allergen challenge. 2. Repeated administration of [Sar(9),Met(O(2))(11)]-substance P ([Sar(9)]SP, 1 microM) maintained mucus output above control values for at least 1.75 h. MEN 11467 inhibited secretion in a concentration-dependent manner with maximal inhibition at 10 microM and an approximate IC(50) of 0.3 microM. Inhibition by MEN 11467 (0.1--10 microM) was maintained, to varying degree, for at least 1.75 h after washout in the continued presence of [Sar(9)]SP. 3. In electrically stimulated tracheae, tachykininergic neural secretion was virtually abolished by 1 microM MEN 11467. 4. In tracheae from ovalbumin-sensitised animals, repeated administration of ovalbumin maintained mucus output above controls for 1.5 h. MEN 11467 inhibited ovalbumin-induced secretion in a concentration-dependent manner, with complete inhibition at 1 microM. Inhibition by MEN 11467 (1 and 10 microM) was maintained, to varying degree, after drug washout for the 1.5 h of ovalbumin stimulation. 5. MEN 11467 1 microM did not affect secretion induced by either acetylcholine or histamine, whereas 10 microM MEN 11467 did inhibit agonist-induced secretion. 6. We conclude that, in ferret trachea in vitro, MEN 11467 at concentrations of 0.1--1 microM is a long acting and selective inhibitor of tachykininergic-induced mucus secretion, and may have therapeutic potential for bronchial hypersecretion associated with allergic conditions, for example in asthma.

Topics: Acetylcholine; Animals; Autonomic Nervous System; Cyclohexylamines; Electric Stimulation; Ferrets; Histamine; Hypersensitivity; Indoles; Male; Mucus; Neurokinin-1 Receptor Antagonists; Ovalbumin; Receptors, Neurokinin-1; Substance P; Sulfur Radioisotopes; Trachea

Asthma may result from excessive Th-2 response in children not previously exposed to Th-1-inducing infections. We tested the hypothesis that BCG vaccination in Th-2-susceptible newborn BP2 mice blocks allergic inflammation and bronchial hyperreactivity (BHR). Ten day-old BP2 mice received 10(5) CFU of BCG 1173P2 intranasally (IN), and 6, 10 or 14 weeks thereafter were sensitized with 100 microg ovalbumin (OVA) in aluminium hydroxide twice subcutaneously (SC) at 1 week interval, and challenged 1 week after the second sensitization with 10 microg OVA IN. Compared to OVA-challenged unvaccinated mice, those that received BCG 8 weeks before challenge developed intense bronchial inflammation, BHR, and high IgE titers. Inflammation involved T cells, macrophages, dendritic cells and was accompanied by increased levels of Interleukin-5 (IL-5) in the bronchoalveolar lavages (BAL). However, animals challenged 16 weeks after BCG vaccination did not develop BHR nor bronchial hypereosinophilia, and showed reduced IgE levels. Bronchial infiltration by immunocompetent cells was also significantly reduced. Increased levels of gamma-interferon (IFN-gamma) after in vitro stimulation of tracheo-bronchial lymph node cells accompanied this blockage, but levels of IL-5 remained high. We demonstrate that 16 weeks after vaccination with BCG in newborn BP2 mice which have a high Th-2 background, allergic inflammation and BHR were blocked, even though a clear Th-1 shift was not achieved.

Topics: Animals; Animals, Newborn; Asthma; BCG Vaccine; Bronchial Hyperreactivity; Child; Cytokines; Dose-Response Relationship, Immunologic; Humans; Hypersensitivity; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Inflammation; Job Syndrome; Lung; Male; Mice; Mycobacterium bovis; Ovalbumin; Phenotype; Th1 Cells; Th2 Cells; Time Factors

Severe respiratory syncytial virus (RSV)-induced disease is associated with childhood asthma and atopy. We combined murine models of allergen-sensitization and RSV infection to explore the interaction of allergic and virus-induced airway inflammation and its impact on airway hyperresponsiveness (AHR). We found that RSV infection during ova-sensitization (OVA/RSV) increased and prolonged AHR compared to mice only RSV-infected (RSV) or ova-sensitized (OVA). AHR is known to be associated with an increase in Type 2 cytokines (IL-4, IL-5, and IL-13) in allergen-sensitized mice. Therefore, we hypothesized that RSV-induced enhancement of AHR was a result of potentiating the Type 2 cytokine profile promoted by ova-sensitization. Surprisingly, we found that Type 2 cytokines induced by ova-sensitization were not increased by RSV infection despite the increase in AHR, and in some cases were diminished. RNAse protection assay revealed no difference in IL-4 and IL-5 mRNA levels between the OVA and OVA/RSV groups, and IL-13 mRNA was significantly decreased in the OVA/RSV mice compared to the OVA group. Flow cytometric analysis of Type 2 cytokines demonstrated the same frequency of IL-4 and IL-5 production in lung-derived T lymphocytes from the OVA/RSV and OVA groups. Direct cytokine ELISA measurements of lung supernatant showed the level of IL-13 was significantly decreased in the OVA/RSV group compared to OVA mice, while there was no difference in either IL-4 or IL-5 between these two groups. These data indicate that the enhanced and prolonged AHR caused by the interaction of allergic airway inflammation and virus-induced immune responses is a complex process that can not be explained simply by augmented production of Type 2 cytokines.

Topics: Allergens; Animals; Cytokines; Disease Models, Animal; Female; Flow Cytometry; Hypersensitivity; Immunization; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; RNA, Messenger; Specific Pathogen-Free Organisms

Chemokine receptors transduce signals important for the function and trafficking of leukocytes. Recently, it has been shown that CC chemokine receptor (CCR)8 is selectively expressed by Th2 subsets, but its functional relevance is unclear. To address the biological role of CCR8, we generated CCR8 deficient (-/-) mice. Here we report defective T helper type 2 (Th2) immune responses in vivo in CCR8(-/)- mice in models of Schistosoma mansoni soluble egg antigen (SEA)-induced granuloma formation as well as ovalbumin (OVA)- and cockroach antigen (CRA)-induced allergic airway inflammation. In these mice, the response to SEA, OVA, and CRA showed impaired Th2 cytokine production that was associated with aberrant type 2 inflammation displaying a 50 to 80% reduction in eosinophils. In contrast, a prototypical Th1 immune response, elicited by Mycobacteria bovis purified protein derivative (PPD) was unaffected by CCR8 deficiency. Mechanistic analyses indicated that Th2 cells developed normally and that the reduction in eosinophil recruitment was likely due to systemic reduction in interleukin 5. These results indicate an important role for CCR8 in Th2 functional responses in vivo.

Topics: Administration, Inhalation; Animals; Antigens; Cockroaches; Cytokines; Dose-Response Relationship, Immunologic; Eosinophils; Granuloma; Hypersensitivity; Immunity, Cellular; Injections, Subcutaneous; Interleukin-5; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Ovum; Receptors, CCR8; Receptors, Chemokine; RNA, Messenger; Schistosoma mansoni; Th1 Cells; Th2 Cells

Eosinophil degranulation is a characteristic feature of asthma and allergic rhinitis. However, degranulated eosinophils have not been convincingly demonstrated in the common mouse models of these airway diseases. This study uses eosinophil peroxidase (EPO) histochemistry and transmission electron microscopy (TEM) analysis to assess eosinophil degranulation in the airways of ovalbumin (OVA)-sensitized and challenged BALB/c and C57BL/6 mice. Using TEM we also examined mouse and human blood eosinophils after in vitro incubation with formyl-Met-Leu-Phe (fMLP) or phorbol myristate acetate (PMA). Although OVA exposure induced significant nasal and lung eosinophilia, we did not observe any of the known cellular processes by which eosinophils release their granule products, i.e., eosinophil cytolysis, piecemeal degranulation, and exocytosis. The occurrence of other allergen-induced degranulation events was ruled out because no difference in granule morphology was observed between lung-tissue eosinophils and blood or bone-marrow eosinophils from control animals. Accordingly, there was no detectable extracellular EPO in lung tissues of allergic mice. Similarly, mouse blood eosinophils remained nondegranulated in vitro in the presence of fMLP and PMA, whereas the same treatment of human eosinophils resulted in extensive degranulation. This investigation indicates that OVA-induced airway inflammation in the present mouse strains does not involve significant eosinophil degranulation. It is speculated that this dissimilarity from the human disease may be due to a fundamental difference in the regulation of mouse and human eosinophils.

Topics: Animals; Cell Degranulation; Cytoplasmic Granules; Disease Models, Animal; Eosinophils; Exocytosis; Humans; Hypersensitivity; In Vitro Techniques; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; N-Formylmethionine Leucyl-Phenylalanine; Nasal Septum; Ovalbumin; Peroxidases; Respiratory Mucosa; Tetradecanoylphorbol Acetate

Asthma and helminth infections induce similar immune responses characterized by the presence of peripheral blood eosinophilia and elevated serum IgE levels. Epidemiological surveys have reported either increases or decreases in the development of atopic diseases and asthma based on the prevalence of helminth infections in the population. The aim of this study was to determine if a pre-existing helminth infection would increase or decrease subsequent allergic responses to an unrelated allergen in the lungs. BALB/cByJ mice were infected with the nematode parasite Strongyloides stercoralis prior to ovalbumin (OVA) immunization and intratracheal challenge. Bronchoalveolar lavage (BAL) and fluid (BALF) were collected 3 days post-challenge and cellular and humoral immune responses were measured. Intracellular cytokine staining revealed increased IL-4 and IL-5 producing cells in BAL from mice infected with S. stercoralis before OVA sensitization. Increased IL-5 protein levels and decreased IFN-gamma protein levels were also observed in the BALF. There was, however, no increase in airway eosinophil accumulation in mice infectd with parasites before sensitization with OVA as compared to mice exposed to OVA alone. Furthermore, eotaxin levels in the lungs induced by OVA was suppressed in mice infected with the parasite before OVA sensitization. The development of OVA specific IgE responses in BALF was also impaired in mice infected with the parasite before sensitization with OVA. These results suggest that a pre-existing helminth infection may potentiate a systemic Type 2-type response yet simultaneously suppress in the lungs allergen-specific IgE responses and eotaxin levels in response to subsequent exposure to allergens.

Topics: Allergens; Animals; Hypersensitivity; Lung; Mice; Ovalbumin; Strongyloides stercoralis; Strongyloidiasis

The influence of stress on total leukocyte count from bronchoalveolar lavage (BAL) was investigated in rats sensitized and challenged with ovalbumin (OVA). The animals were injected intraperitoneally with a suspension of OVA plus aluminum hydroxide in 0.9% NaCl (Day 0) and boosted at Day 7 with an identical OVA solution, administered subcutaneously. From the first to the 13th day after sensitization, rats were placed individually in a shuttle box where they received 50 escapable footshocks per day, always preceded by a sound signal (S); the responses that occurred during both S and shocks canceled the stressful stimulation. On Day 14, animals were submitted to a single session of 50 inescapable footshocks, preceded by the same S; immediately after, the animals were submitted to a 1% OVA-inhalation challenge. Results showed high levels of stress in the shocked animals as detected through both ultrasonic vocalizations (UVs) and social interaction test in an open field. Total leukocyte count in BAL from stressed animals (24 h post-OVA challenge) revealed a significant increase in the number of inflammatory cells in comparison to that measured in sensitized, nonstressed challenged rats. These data demonstrate that stress plays a relevant and important role on total bronchoalveolar cell count in OVA-sensitized rats.

Topics: Animals; Bronchoalveolar Lavage Fluid; Cell Count; Electroshock; Hypersensitivity; Interpersonal Relations; Leukocyte Count; Male; Ovalbumin; Rats; Rats, Wistar; Stress, Psychological; Vocalization, Animal

Many clinical and experimental data have shown that learning can occur under general anesthesia. To clarify this possibility with respect to allergic reactions, particularly asthmatic responses, we first established classical conditioned histamine release in response to a neutral odor by using pairings of the odor and an inhaled antigen for five sessions (Experiment 1) and then investigated whether conditioned histamine release into the plasma, bronchoalveolar lavage fluid (BALF), and lung tissue, which followed such a conditioning procedure, would be produced in urethane-anesthetized guinea pigs in the presence or absence of antigen (Experiment 2). Ovalbumin (OA) was used as the unconditioned stimulus (US) and dimethylsulfide (DMS) served as the conditioned stimulus (CS) in both experiments. In Experiment 1, the plasma histamine levels in the conditioned group increased significantly more than those of the unpaired control group in response to the CS during consciousness. In Experiment 2 in the absence of antigen, however, no significant differences in the histamine levels were found regarding the groups (DMS, triethylamine, saline, or unsensitized) or the time course (before, immediately, 5 min, and 10 min after the inhalations) during anesthesia, except for the finding that the histamine levels in the lung tissue specimens from the DMS group were significantly higher than those from the triethylamine group. In Experiment 2 in the presence of antigen, there was a significant increase in the plasma histamine levels after exposure to the US, irrespective of the presence of the CS, however, no significant difference in the histamine levels was observed between the US and the CS+US groups. These results indicated that a classically CS might not induce asthmatic responses under anesthesia.

Topics: Anesthesia, Intravenous; Anesthetics, Intravenous; Animals; Conditioning, Psychological; Guinea Pigs; Histamine; Histamine Release; Hypersensitivity; Lung; Male; Odorants; Ovalbumin; Passive Cutaneous Anaphylaxis; Urethane

T lymphocytes play a critical role in the development of allergic inflammation in asthma. Early in the allergic response, T lymphocytes migrate from the circulation into the lung to initiate and propagate airway inflammation. The adhesion molecules that mediate lymphocyte entry into inflamed lung have not been defined. This study directly examined the roles of L-selectin and intercellular adhesion molecule 1 (ICAM-1) in lymphocyte migration to the lung during an allergic inflammatory response in an animal model of asthma. Short-term (1 hour) in vivo migration assays and various combinations of adhesion molecule-deficient and wild-type mice were used. Migration of in vivo activated lymphocytes into inflamed lung was significantly greater than entry of resting lymphocytes into noninflamed lung (24.5% +/- 2.7% vs 9.5% +/- 1.3%, P =.001). Migration of activated lymphocytes into inflamed lung was inhibited by 30% in the absence of L-selectin (17.3% +/- 1.3%, P =.04), 47% in the absence of cell surface ICAM-1 (13.0% +/- 2.5%, P =.01), and 47% in the absence of endothelial ICAM-1 (13.0% +/- 2.5%, P =.01). Loss of ICAM-1 on both lymphocytes and lung endothelium inhibited lymphocyte migration by 60% (9.8% +/- 1.8%, P =.002). These findings demonstrate clear roles for both L-selectin and ICAM-1 in lymphocyte migration to the lung during an allergic inflammatory response, with ICAM-1 playing a greater role.

Topics: Animals; Asthma; Cell Movement; Disease Models, Animal; Hypersensitivity; Inflammation; Intercellular Adhesion Molecule-1; L-Selectin; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocytes

p59fynT is a protein tyrosine kinase in the src family that has been associated with and believed to function in the signaling of many receptors, including the T-cell receptor. A role for the kinase in antigen-driven pulmonary inflammation was examined using mice whose p59fynT gene had been genetically ablated. FynKO mice that were sensitized to ovalbumin exhibited a marked increase in bronchoalveolar lavage eosinophils and cytokines, including interleukin (IL)-4 and IL-5, relative to wild-type mice in response to antigen aerosol exposure. Ovalbumin-stimulated IL-5 production was also increased in cultured splenocytes derived from fynKO mice relative to wild-type mice, whereas interferon-gamma levels were unchanged. Diminished concanavalin A--stimulated IL-4 levels from fynKO splenocytes were consistent with reduced serum immunoglobulin (Ig)E levels observed in sensitized/saline aerosol-challenged animals and may reflect defective natural killer 1.1(+) T cell development. Normalization of IgE levels in sensitized fynKO mice relative to wild-type mice occurred after repeat antigen challenge, which suggests a secondary source of IL-4. Overall, these data demonstrate fyn is a negative regulator of allergic airway inflammation in mice because its absence promotes a shift to a T helper-2 phenotype that may reflect the kinase's role in T-cell receptor signaling.

Topics: Animals; Antigens; Bronchoalveolar Lavage Fluid; Concanavalin A; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunoglobulin E; Interleukin-4; Interleukin-5; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pneumonia; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-fyn; Spleen; src-Family Kinases; Th2 Cells

Airway inflammation and airway hyperresponsiveness (AHR) are fundamental features of asthma. Migration of inflammatory cells from the circulation into the lungs is dependent upon adhesion molecule interactions. The cell surface adhesion molecules L-selectin and intercellular adhesion molecule (ICAM)-1 have been demonstrated to mediate leukocyte rolling on inflamed pulmonary endothelium, and ICAM-1 has also been shown to mediate capillary sequestration in inflamed lung. However, their roles in the development of airway inflammation and AHR in asthma have not been directly examined. We have characterised the roles of L-selectin and ICAM-1 in the recruitment of inflammatory cells to the lung and in the development of airway hyperresponsiveness using an ovalbumin (OVA)-induced allergic airway disease model of asthma and adhesion molecule-deficient mice. OVA-sensitized/challenged ICAM-1-deficient mice have dramatically reduced inflammatory influx into the airway/lung and a corresponding attenuation of AHR as compared to wild-type controls. OVA-sensitized/challenged L-selectin-deficient mice demonstrate significantly reduced numbers of CD3(+)lymphocytes and increased numbers of B220(+)lymphocytes in BAL as compared to wild-type mice (P< 0.05). However, other parameters of airway/lung inflammation in OVA-sensitized/challenged L-selectin-deficient mice were equivalent to wild-type control mice. Remarkably, despite a fulminant inflammatory response in the airway/lung, AHR was completely abrogated in OVA-sensitized/challenged L-selectin-deficient mice. These findings suggest a crucial role for ICAM-1 in the development of airway inflammation and AHR in asthma. In contrast, L-selectin plays a more selective role in the development of airway hyperresponsiveness but not allergic inflammation in this animal model of asthma. Thus, L-selectin and ICAM-1 represent potential targets for novel asthma therapies specifically aimed at controlling airway inflammation and/or airway hyperresponsiveness.

Topics: Airway Resistance; Animals; Asthma; Cell Movement; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Intercellular Adhesion Molecule-1; L-Selectin; Male; Mice; Ovalbumin

We studied the conditions needed to sensitize animals to the oral feeding of food allergens, without induction of tolerance, in order to investigate the allergenicity of orally ingested food proteins. Brown Norway (BN) rats were sensitized by daily OVA (ovalbumin)-gavage or by drinking OVA containing water ad libitum and the ASA (active systemic anaphylaxis) response, as the immediate hypersensitivity response to antigen stimulation after oral sensitization, was examined. The oral administration of OVA by gavage produced a higher OVA-specific IgE response and an increase in serum histamine after antigen challenge, as compared to those produced by drinking water. Next, we examined the effect of murine age, the oral feeding technique and the oral feeding dose on sensitization using BALB/c, B10A and ASK mice. Twenty-week-old mice showed the strongest OVA-specific IgE and IgG1 responses and ASA-associated serum histamine contents increased with gavage in the three different age groups of BALB/c mice. Administering 0.1 mg of OVA by gavage daily for 9 weeks appeared to induce a higher response than administering 1 mg of OVA, in terms of OVA-specific IgE and IgG1 antibody responses and ASA responses. Among the three strains of mice, B10A mice exhibited the highest response in terms of OVA-specific IgE and IgG1 antibody and ASA responses. These findings suggested BN rats and B10A mice were suitable models for oral sensitization with antigen protein and that oral sensitization in mice requires low dose, intermittent antigen intakes.

Topics: Administration, Oral; Age Factors; Allergens; Anaphylaxis; Animals; Disease Models, Animal; Female; Histamine; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Rats, Inbred BN; Species Specificity; Time Factors

We investigated the presence of mast cell granules in macrophages following an in vivo model of an allergic reaction. Injection of ovalbumin (100 microg) into the peritoneal cavity of sensitised mice produced a rapid (within 2 h) influx of neutrophils followed by a slower (after >4 h) eosinophil migration. Ovalbumin treatment induced a high incidence (approximately 50%) of mast cell degranulation compared to control phosphated-buffered saline-treated mice. The majority (approximately 90%) of peritoneal macrophages contained mast cell granules as early as 2 h post-ovalbumin, with lower values at later time-points, as determined by staining with Toluidine blue and Berberine sulphate. This was confirmed by electron microscopy which enabled us to identify the complex mast cell granule sub-structural components in macrophage phagosomes. In conclusion, we used histochemical and ultrastructural analyses to show that mast cell granules become internalised with macrophages during the early stages of an experimental allergic reaction.

Topics: Animals; Cell Degranulation; Cell Movement; Female; Hypersensitivity; Macrophages, Peritoneal; Mast Cells; Mice; Mice, Inbred BALB C; Microscopy, Electron; Ovalbumin; Peritoneal Cavity; Secretory Vesicles; Time Factors

Because interferon-gamma, interleukin-4, and interleukin-5 have been identified at the mRNA and protein levels in the lesional skin of patients with atopic dermatitis, we investigated the roles played by granulocytes as effector cells in allergic inflammation by using two unique murine skin models. In vitro generated Th1 and Th2 cells from naïve splenocytes of antiovalbumin T cell receptor transgenic BALB/C mice were adoptively transferred with ovalbumin into the ear pinnae or air-pouches produced in the back skin of naïve, nontransgenic BALB/C mice. The injection of Th1 cells with ovalbumin induced delayed type ear swelling that peaked at 48 h, whereas that of Th2 resulted in ear swelling that peaked at a much earlier time, 24 h. Histologic study of the swollen ear skin and granulocytes recruited into the air-pouch demonstrated that, although the Th1-induced inflammation caused a neutrophil-predominant infiltrate with few eosinophils, larger numbers of eosinophils accumulated in the Th2-induced inflammation. Using these murine models, we further evaluated the effects of drugs used for the treatment of atopic diseases. The results showed that FK506 administration could effectively reduce skin inflammation induced by either Th cells. Interestingly, the neutrophil elastase inhibitor ONO-6818 efficiently inhibited Th1-induced inflammation. In contrast, a leukotriene receptor antagonist, ONO-1078, specifically suppressed Th2-induced inflammation. We also found that each ONO drug exerted direct influence on specified granulocytes, as neither affected in vitro production of relevant Th cytokines. Thus, we succeeded in developing animal skin inflammation models in which we can evaluate the contribution of protein antigen-specific Th1 or Th2 cells through the action of granulocytic effector cells.

Topics: Animals; Cells, Cultured; Chromones; Dermatitis, Atopic; Disease Models, Animal; Ear; Edema; Enzyme Inhibitors; Eosinophils; Hypersensitivity; Immunosuppressive Agents; Leukotriene Antagonists; Male; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Oxadiazoles; Pyrimidinones; Skin; Tacrolimus; Th1 Cells; Th2 Cells

We detected a human humoral peptide that deglycosylates mouse antibody IgE, and found that this peptide had the amino acid sequence ADSGEGDFLAEGGGV. The peptide synthesized according to the sequence also deglycosylated the antibody IgE. The deglycosylated IgE did not induce mouse systemic anaphylaxis. Human fibrinopeptide A has the amino acid sequence indicated above, and a polypeptide extracted from human urine showed an antiallergic effect on humans and mice, which strongly suggests that fibrinopeptide A mediates allergic reaction via antibody IgE-deglycosylation, and is excreted as a polypeptide in urine.

Topics: Acute-Phase Reaction; Adult; Amino Acid Sequence; Animals; Drug Hypersensitivity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Fibrinopeptide A; Glycosylation; Glycosyltransferases; Humans; Hypersensitivity; Immune Sera; Immunoglobulin E; Injections, Intraperitoneal; Male; Mice; Mice, Inbred Strains; Molecular Sequence Data; Ovalbumin; Peptide Fragments; Structure-Activity Relationship

Although it is not proven, one factor considered important in the development of sinusitis is allergic rhinitis.. The purpose of this study was to determine whether ongoing allergic rhinitis enhances the infection and inflammation associated with Streptococcus pneumoniae acute sinus infection.. BALB/c mice were sensitized to ovalbumin by intraperitoneal injection. After infection of the sinuses by S pneumoniae, either with or without concomitant administration of ovalbumin to induce allergic inflammation, mice were killed at various times and their heads were prepared for histologic evaluation of the sinuses.. Mice became allergic to ovalbumin and developed eosinophilia in the sinus and lung cavities in response to ovalbumin administration to each of the respective cavities. In comparison with controls, the mice with ongoing nasal allergic inflammation that were inoculated with S pneumoniae had significantly more bacteria recovered at sacrifice and had significantly more inflammation, as indicated by neutrophil, eosinophil, and mononuclear influx into the sinus mucosa. The percentage of the sinus area occupied by neutrophil clusters was also increased after infection in the allergic mice in comparison with the control mice.. Our data demonstrate that mice can be sensitized to ovalbumin and develop a localized allergic reaction in the skin, nose, or lung. An ongoing local allergic response augments bacterial infection in these animals. We also demonstrate that allergic sensitization alone, allergen exposure alone, or an allergic response at a distal site, the lung, does not augment the sinus infection.

Topics: Animals; Eosinophils; Female; Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Mucous Membrane; Neutrophils; Ovalbumin; Pneumococcal Infections; Sinusitis

1. The contribution of reactive nitrogen species to the development of airway hyperresponsiveness in a mouse model of allergic inflammation was investigated by the use of selective inhibitors of nitric oxide and superoxide formation. 2. Sensitized mice, repeatedly challenged with ovalbumin showed a significant (P<0.001, n=9) increase in airway responsiveness measured using whole body plethysmography. This hyperresponsiveness was accompanied by an influx of eosinophils into the airway lumen and increased levels of ovalbumin-specific serum IgE. 3. Treatment of mice with the iNOS inhibitor 1400 W or the NADPH-oxidase inhibitor apocynin did not significantly alter cellular influx into the airway lumen nor serum ovalbumin specific IgE. In contrast, apocynin as well as 1400 W inhibited ovalbumin-induced airway hyperresponsiveness (P<0.001 and P<0.05 respectively, n=9). Furthermore, the airways of allergen challenged animals showed clear 3-nitrotyrosine staining, which was mainly located in eosinophils. Remarkably, treatment with apocynin or 1400 W did not alter 3-nitrotyrosine staining. 4. These data suggest that the development of airway hyperresponsiveness during the airway inflammation upon ovalbumin challenge is dependent on the release of both superoxide and nitric oxide and is therefore likely to be dependent on reactive nitrogen species. This mechanism, however, is not reflected by 3-nitrotyrosine formation in the airways.

Topics: Acetophenones; Amidines; Animals; Antioxidants; Benzylamines; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Enzyme Inhibitors; Eosinophils; Hypersensitivity; Immunoglobulin E; Immunohistochemistry; Interferon-gamma; Interleukin-4; Interleukin-5; Lung; Male; Mice; Mice, Inbred BALB C; Neutrophils; Nitric Oxide Synthase; Ovalbumin; Specific Pathogen-Free Organisms; Tyrosine

Although the levels of immunoglobulin E (IgE) in the circulating blood are often elevated in patients with allergic diseases, such levels cannot always be considered as pathognomonic signs of allergy. The induction of allergic reactions in the tissue was inferred to be related to the amount of IgE passing through the vascular wall.. We attempted to clarify which compartment, the intravascular or extravascular, plays an important role in the regulation of the turnover of rat IgE.. The level of DNP-specific rat IgE in the serum was estimated by IgE-capture enzyme-linked immunosorbent assay, and the turnover of IgE was analyzed from its pharmacokinetic parameters.. The transfer rate constants from the central to tissue compartment (Kct) were larger than those from the tissue to central compartment (Ktc) irrespective of the sensitized state. The value of the distribution volume of the tissue compartment (Vt) was larger than that of the distribution volume of the central compartment (Vc) irrespective of the sensitized state.. These Findings suggest that the short half-life of rat IgE in the circulation could be attributable to the distribution of IgE from the intravascular to the extravascular compartment.

Topics: Animals; Antibodies, Monoclonal; Benzenesulfonates; Body Fluid Compartments; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Humans; Hypersensitivity; Immunoglobulin E; Male; Ovalbumin; Rats; Rats, Wistar

Leptin is important for maintenance of the body's energy homeostasis and it also increases Th1 and suppresses Th2 cytokine production. We have investigated the effect of leptin on the allergic immune response to the model allergen ovalbumin (OA) by using the popliteal lymph node assay (PLNA) and serum antibody determination in mice. Mice were injected with either leptin i.v. plus OA in one hind footpad, or leptin or OA alone. A booster dose of leptin was given twice and of OA once and the animals were exsanguinated on experimental day 19 when the PLNs also were removed. End-point measurements were serum levels of IgE, IgG1, and IgG2a anti-OA and weight and cell number of the excised PLNs. Leptin given i.v. with the protocol employed altered neither the cellular PLN response nor the specific serum IgE, IgG1, or IgG2a anti-OA levels compared with the group given OA without leptin. Our data indicate that systemic administration of leptin neither suppresses nor enhances the Th2-dependent antibody responses in the present mouse model.

Topics: Allergens; Animals; Cells, Cultured; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Leptin; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th2 Cells

To examine the differential properties of mucous glycoproteins, we produced hypertrophic and metaplastic changes in goblet cells of rat nasal epithelium by intranasal instillation of ovalbumin (OVA) in OVA-sensitized rats, and by intranasal lipopolysaccharide (LPS) instillation. The epithelial mucosubstance was quantitatively examined by alcian blue-periodic acid-Schiff (AB-PAS) and lectin histochemistry. The newly produced mucin after OVA challenge or LPS instillation contained a high amount of sulfomucin and a low amount of neutral glycoprotein: LPS-induced mucin contained more sulfomucin (70.1% of total) and less neutral glycoprotein (8.6%) than OVA-induced mucin (sulfomucin, 33.6%; neutral glycoprotein, 41.8%; p < 0.01). Four of the lectins stained some of the mucosubstance, indicating the presence of galactose-N-acetylgalactosamine, alpha2,3- and alpha2,6-linked sialic acid-galactose, and fucose residues. After LPS instillation, the reactivity was higher for galactose-N-acetylgalactosamine (64.8% of total) and alpha2,3-linked sialic acid-galactose (75.8%) than after saline instillation (3.5 and 19.1%, respectively) or OVA challenge (5.8 and 32.3%; p < 0.05). OVA challenge did not induce the alteration of terminal sugar residues. A 2-fold increase in mucin mRNA (rat Muc5ac) expression was induced after LPS instillation or OVA challenge, compared with animals treated with saline instillation (p < 0.05). These results indicate that mucin mRNA expression (for peptide backbone) increases similarly after LPS instillation or OVA challenge; however, carbohydrate compositions of newly produced mucin are different between the two groups.

Topics: Animals; Data Interpretation, Statistical; Epithelium; Goblet Cells; Histocytochemistry; Hypersensitivity; Inflammation; Lectins; Lipopolysaccharides; Male; Mucins; Mucus; Ovalbumin; Rats; Rats, Inbred F344; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sodium Chloride

We attempted to elucidate the contribution of complement to allergic asthma. Rat sensitized to OVA received repeated intratracheal exposures to OVA for up to 3 consecutive days, and pulmonary resistance was then estimated for up to 6 h after the last exposure. Whereas the immediate airway response (IAR) in terms of R(L) tended to decrease in proportion to the number of OVA exposures, late airway response (LAR) became prominent only after three. Although premedication with two kinds of complement inhibitors, soluble complement receptor type 1 (sCR1) or nafamostat mesylate, resulted in inhibition of the IAR after either a single or a double exposure, the LAR was inhibited after the triple. Premedication with a C5a receptor antagonist (C5aRA) before every exposure to OVA also inhibited the LAR after three. Repeated OVA exposure resulted in eosinophil and neutrophil infiltration into the bronchial submucosa which was suppressed by premedication with sCR1 or C5aRA. Up-regulation of C5aR mRNA was shown in lungs after triple OVA exposure, but almost no up-regulation of C3aR. Pretreatment with sCR1 or C5aRA suppressed the up-regulation of C5aR expression as well as cytokine messages in the lungs. The suppression of LAR by pretreatment with sCR1 was reversed by intratracheal instillation of rat C5a desArg the action of which was inhibited by C5aRA. In contrast, rat C3a desArg or cytokine-induced neutrophil chemoattractant-1 induced cellular infiltration into the bronchial submucosa by costimulation with OVA, but these had no influence on the LAR. These differences might be explained by the fact that costimulation with OVA and C5a synergistically potentiated IAR, whereas that with OVA and either C3a or cytokine-induced neutrophil chemoattractant-1 did not. C5a generated by Ag-Ab complexes helps in the production of cytokines and contributes to the LAR after repeated exposure to Ag.

Topics: Airway Resistance; Animals; Antigens; Antigens, CD; Asthma; Benzamidines; Bronchi; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokines, CC; Chemokines, CXC; Chemotactic Factors; Complement C3a; Complement C5a; Complement C5a, des-Arginine; Cytokines; Growth Substances; Guanidines; Hypersensitivity; Intercellular Signaling Peptides and Proteins; Lung; Membrane Proteins; Ovalbumin; Rats; Receptor, Anaphylatoxin C5a; Receptors, Complement; Receptors, Complement 3b; RNA, Messenger

Epidemiological studies have suggested increased prevalence of atopy in children of maternal smokers. Although secondhand smoke or environmental tobacco smoke (ETS) has been shown to augment allergic responses, its role in atopic sensitization is still controversial. We studied whether ETS could initiate a Th2 response and thus induce primary allergic sensitization. Mice were exposed for 10 consecutive days to either 1% aerosolized OVA, ETS (5 cigarettes), or both ETS and OVA. C57BL/6 mice receiving both ETS and OVA developed OVA-specific IgE and IgG1, 12, 14, and 25 days after the initial exposure, whereas those receiving OVA alone did not. Thirty days after the initial challenge (20 days after its completion), mice were re-exposed to OVA. Bronchoalveolar lavage performed 24 h later revealed an influx of eosinophils in the group initially challenged with both ETS and OVA, but not in those exposed to ETS alone or OVA alone. Increases in IL-5, GM-CSF, and IL-2 were observed in bronchoalveolar lavage from this OVA/ETS-exposed group, whereas IFN-gamma levels were significantly inhibited. These results suggest that ETS can induce allergic sensitization to a normally harmless Ag, and they may explain why secondhand smoke is a major risk factor for the development of allergy in children.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Pulmonary Eosinophilia; Species Specificity; Tobacco Smoke Pollution

Severe respiratory syncytial virus (RSV) infection has been hypothesized to be a risk factor for the development of allergy and asthma, but epidemiologic studies in humans have been inconclusive. By use of a well-characterized murine model of RSV infection and allergic sensitization with ovalbumin, the effect of a preceding severe RSV infection on the development of the pulmonary allergic inflammatory response and airway hyperresponsiveness (AHR) was tested. The impact of prior allergic sensitization on RSV-induced illness, as measured by weight loss, also was evaluated. RSV infection before allergic sensitization decreased allergen-induced AHR, production of interleukin-13 in lung tissue, and lung eosinophilia. In contrast, allergic sensitization before RSV infection increased AHR and decreased RSV-related weight loss and lung levels of interferon-gamma but did not alter viral clearance. These data provide evidence that RSV-associated AHR occurs in hosts with allergic responses and that allergic inflammation is diminished when preceded by RSV infection.

Topics: Allergens; Animals; Bronchial Hyperreactivity; Bronchoconstrictor Agents; Female; Hypersensitivity; Interferon-gamma; Interleukin-13; Lung; Lymphocyte Count; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Time Factors; Virus Replication; Weight Loss

Interleukin (IL)-9 is a T-cell-derived cytokine with pleiotropic activities on T helper 2 cells, B cells, and mast cells. IL-9 may therefore play an important role in the development of allergic pulmonary inflammatory diseases. In this study, an antimouse IL-9 (anti-mIL-9) antibody (Ab) was evaluated against pulmonary eosinophilia, histopathologic changes in lung tissues, serum immunoglobulin (Ig) E levels, and airway hyperresponsiveness (AHR) to methacholine in mice sensitized and challenged with ovalbumin (OVA). Additionally, steady-state levels of IL-4, IL-5, IL-13, and interferon-gamma messenger RNA (mRNA) in the lungs were measured. The anti-mIL-9 Ab (200 microg/mouse, intraperitoneally) was given as either four doses during the sensitization period or as a single dose before OVA challenge. Sensitized mice challenged with OVA displayed marked pulmonary eosinophilia, epithelial damage, and goblet cell hyperplasia. OVA challenge also increased mRNA levels of IL-4, IL-5, and IL-13 in the lungs. AHR was also increased twofold in sensitized, challenged mice. Treatment of sensitized, challenged mice with four doses of anti-mIL-9 Ab significantly reduced pulmonary eosinophilia, serum IgE levels, goblet cell hyperplasia, airway epithelial damage, and AHR, but had no effect on IL-4, IL-5, and IL-13 mRNA levels in the lungs. A single dose of the antibody was ineffective on all measures. These results indicate that an antibody to mIL-9 inhibits the development of allergic pulmonary inflammation and AHR in mice.

Topics: Animals; Antibodies, Monoclonal; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Eosinophils; Gene Expression; Goblet Cells; Hypersensitivity; Immunoglobulin E; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Interleukin-9; Male; Mice; Mice, Inbred Strains; Ovalbumin; Pneumonia; Respiratory Mucosa; RNA, Messenger

The ovalbumin (OVA)-sensitized guinea pig is often used as an animal model of asthma and airway hyperreactivity. A characteristic lesion of asthma is excessive production of mucin in the airways. Mechanistic studies of this lesion in guinea pigs have been limited due to lack of mucin gene probes for this species. The aim of the present study was to clone the cDNAs encoding two major airway mucins (Muc2 and Muc5ac) from the guinea pig, and investigate mucin gene expression in lungs of sensitized animals in response to antigen challenge. We isolated and sequenced two cDNA fragments coding for the sequences located within the carboxyl-terminal cysteine-rich region of guinea pig Muc2 and Muc5ac mucins. Comparison of cloned cDNAs with those from other species revealed high degrees of sequence identity and conservation of all cysteine residues in deduced primary sequences. Based on the resultant sequence information, we also designed oligonucleotide primers for specific detection of guinea-pig Muc2 and Muc5ac steady-state mRNA levels via reverse transcriptase/ polymerase chain reaction (RT-PCR). Levels of both Muc2 and Muc5ac mRNA in lungs of OVA-sensitized guinea pigs increased significantly by 30 min after an acute exposure to 0.3% OVA. In addition, levels of eotaxin mRNA also increased in these tissues, but the increases were not significant until 2 h after challenge. Correspondingly, the number of eosinophils in bronchoalveolar lavage fluid did not increase until 4 h postchallenge. Results of these studies suggest that the OVA-sensitized guinea pig responds to allergic challenge with enhanced expression of genes (e.g., eotaxin, Muc2, and Muc5ac) that likely play a role in increased airway inflammation and mucin overproduction, and enhanced mucin gene expression appears to occur before eosinophil infiltration.

Topics: Amino Acid Sequence; Animals; Antineoplastic Agents; Asthma; Base Sequence; Bronchial Hyperreactivity; Cells, Cultured; Chemokine CCL11; Chemokines, CC; Cloning, Molecular; Conserved Sequence; Disease Models, Animal; Eosinophils; Gene Expression; Guinea Pigs; Histamine; Hypersensitivity; Inflammation Mediators; Molecular Sequence Data; Mucin 5AC; Mucin-2; Mucins; Ovalbumin; Respiratory Mucosa; RNA, Messenger; Trachea; Tumor Necrosis Factor-alpha

Some particulate matter is known to affect human health, yet the mechanism(s) by which it acts is largely unknown. One of the factors that may play a role in the immune- stimulating activity of particles is binding of allergen to particles. This may turn the particles into allergen carriers, resulting in antigen deposition within the altered inflammatory microenvironment created by the particles. We compared the efficacy of simultaneous versus separate administration of antigen and particles during sensitization in an intranasal exposure model in BALB/c mice. Sensitization consisted of three separate doses (10 microg) of TNP-OVA at Days 1, 2, and 3. Two hundred micrograms of carbon black particles (CBP) were administered either 1 day before sensitization (Day 0), 1 day after sensitization (Day 4), or during sensitization. The latter was performed either at Day 1 (200 microg) or at Days 1, 2, and 3 (67 microg/day). At Day 10 a challenge with 10 microg of TNP-OVA was performed, and at Day 15 the immune response was assessed. The total number of cells as well as antibody-forming cells (AFC) in lymph nodes draining the lung (peribronchial lymph nodes [PBLN]) were determined, and immunoglobulin levels in blood were assessed. Cell numbers of PBLN increased significantly in all particle-treated groups compared to controls. The number of TNP-specific IgG1-forming cells in the groups receiving particles during sensitization was significantly higher than control level. Only groups receiving particles during or before sensitization displayed significantly higher IgG1 levels than controls, in contrast to the group receiving particles after sensitization. Only in animals receiving three doses of 67 microg during sensitization did TNP-specific IgE increase significantly compared to controls. IgG2a did not show significant differences compared to controls, indicating that the response is predominantly Th2 mediated. These data indicate that coadministration of particles at all time points of antigen dosing is the most effective way to stimulate an immune response in our model compared to separate particle and antigen dosing. Also, administration shortly before antigen administration was effective in stimulating an immune response, suggesting that time-dependent processes are involved in immune-stimulating activity of particles, supporting the important role of the altered inflammatory microenvironment created by the particles.

Topics: Administration, Intranasal; Animals; Antibody-Producing Cells; Carbon; Cell Count; Disease Models, Animal; Drug Administration Schedule; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Time Factors

Replication-deficient adenovirus vector (Ad) is one of the most efficient gene transfer vehicles for human gene therapy. However, Ad is antigenic, known to evoke prominent inflammatory responses in vivo, and there are concerns that using Ad in patients with immune-mediated disorders (allergy and autoimmune diseases) may affect the status of the diseases. To evaluate this concept in a manner close to clinical scenarios, a mouse model of airway eosinophilic inflammation was developed by administering intraperitoneal injections and inhalations of chicken ovalbumin (OA), with Ad administered intranasally 5 days after the OA sensitization. The administration of Ad resulted in a significant suppression of eosinophil counts in peripheral blood as well as in the bronchoalveolar lavage fluid (BALF), and a decrease in OA-specific IgE. The decrease in the number of eosinophils in BALF was associated with a marked upregulation of interferon gamma (IFN-gamma) expression. In contrast, the Ad-specific, delayed-type hypersensitivity response and efficacy of reporter gene expression mediated by Ad were only marginally affected in animals sensitized with OA. Together, these data support the idea that Ad administration in patients with Th2-mediated immune disorders does not exacerbate the parameters of ongoing inflammations or gene transfer efficiency, and with its ability to induce prominent type 1 immune response to the antigen in vivo, Ad could potentially be used as an efficient adjuvant to control immune disorders where Th2 cell-mediated mechanisms are involved.

Topics: Adenoviridae; Animals; Bronchoalveolar Lavage Fluid; Chickens; Eosinophils; Gene Transfer Techniques; Genes, Reporter; Genetic Vectors; Humans; Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulin E; Inflammation; Interferon-gamma; Leukocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Despite marked advances in the understanding of allergic responses, the mechanisms regulating gastrointestinal allergy are not very well understood. We have developed a model of antigen-induced eosinophil-associated gastrointestinal allergy and characterized the role of eotaxin and IL-5. Challenge of allergen-sensitized mice with oral allergen, in the form of enteric-coated beads, resulted in marked allergen-specific IgG(1) and IgE, Th(2)-type (IL-4 and IL-5) cytokine production, and eosinophil accumulation in the blood and small intestine. In the genetic absence of eotaxin, a chemokine constitutively expressed in the gastrointestinal tract, eosinophil recruitment into the small intestine was ablated, and these mice developed enhanced eosinophil accumulation in the blood compared with wild-type mice. Interestingly, in the absence of IL-5, allergen challenge promoted partial eosinophil accumulation into the small intestine and a decline in circulating eosinophil levels. Collectively, these results establish that the accumulation of gastrointestinal eosinophils is antigen induced, can occur independent of IL-5, and provides a molecular mechanism to explain the dichotomy between peripheral blood and tissue eosinophilia. Furthermore, eotaxin is identified as a critical regulator of antigen-induced eosinophilic inflammation in the gastrointestinal tract.

Topics: Anaphylaxis; Animals; Chemokine CCL11; Chemokines, CC; Chemotactic Factors, Eosinophil; Cytokines; Eosinophils; Female; Gastrointestinal Diseases; Hypersensitivity; Integrins; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, CCR3; Receptors, Chemokine

Macrophage migration inhibitory factor (MIF) has recently been forwarded as a critical regulator of inflammatory conditions, and it has been hypothesized that MIF may have a role in the pathogenesis of asthma and chronic obstructive pulmonary disease (COPD). Hence, we examined effects of MIF immunoneutralization on the development of allergen-induced eosinophilic inflammation as well as on lipopolysaccharide (LPS)-induced neutrophilic inflammation in lungs of mice. Anti-MIF serum validated with respect to MIF neutralizing capacity or normal rabbit serum (NRS) was administered i.p. repeatedly during allergen aerosol exposure of ovalbumin (OVA)-immunized mice in an established model of allergic asthma, or once before instillation of a minimal dose of LPS into the airways of mice, a tentative model of COPD. Anti-MIF treatment did not affect the induced lung tissue eosinophilia or the cellular composition of bronchoalveolar lavage fluid (BALF) in the asthma model. Likewise, anti-MIF treatment did not affect the LPS-induced neutrophilia in lung tissue, BALF, or blood, nor did it reduce BALF levels of tumor necrosis factor-alpha (TNF-alpha) and macrophage inflammatory protein-1alpha (MIP-1alpha). The present data suggest that MIF is not critically important for allergen-induced eosinophilic, and LPS-induced neutrophilic responses in lungs of mice. These findings do not support a role of MIF inhibition in the treatment of inflammatory respiratory diseases.

Topics: Animals; Dose-Response Relationship, Drug; Eosinophils; Humans; Hypersensitivity; Lipopolysaccharides; Macrophage Migration-Inhibitory Factors; Macrophages; Mice; Mice, Inbred C57BL; Models, Immunological; Neutrophils; Ovalbumin; Pneumonia; Rabbits; Trachea

To determine the impact of bacillus Calmette Guerin (BCG) vaccination on IgE production in ovalbumin (OVA)-sensitized newborn mice, four groups (I, II, III, IV) of BALB/c mice were immunized on the first day of life with live BCG, killed BCG, BCG diluent, and saline, respectively. No injection was applied to mice in group V (control). All mice except group V were sensitized and challenged with OVA in the fourth and sixth weeks, respectively, and serum total IgE levels were determined at 8 weeks, 2 weeks after the second OVA challenge. IgE levels of all groups were significantly higher than the control group except for group II (p = 0.95). Mice in group II showed significantly lower IgE values than group IV and I (p = 0.007 and p = 0.003, respectively). We concluded that heat-killed BCG may downregulate IgE response to OVA in newborn mice.

Topics: Animals; Animals, Newborn; BCG Vaccine; Down-Regulation; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mycobacterium bovis; Ovalbumin; Pregnancy; Vaccines, Attenuated; Vaccines, Inactivated

The immunomodulatory role of arachidonic acid metabolites in allergic sensitization is undefined. Prostaglandin E(2) (PGE(2)), a product of arachidonic acid metabolism through the cyclooxygenase pathway, has been reported to favor Type 2-like cytokine secretion profiles in murine and human CD4(+) T cells by inhibiting the production of Type 1-associated cytokines. On the basis of these in vitro data, we hypothesized that indomethacin, a nonselective cyclooxygenase inhibitor, would diminish allergen-induced production of Type 2 cytokines in mice, and protect against airway hyperresponsiveness (AHR) to methacholine. We found that ovalbumin-sensitized mice that were treated with indomethacin (OVA-indomethacin mice) had significantly greater AHR (p < 0.05) and higher levels of IL-5 (176 +/- 52 versus 66 +/- 4 pg/ml) and IL-13 (1,226 +/- 279 versus 475 +/- 65 pg/ml) in lung supernatants than mice sensitized with ovalbumin alone (OVA mice), while levels of IL-4 and serum IgE were not different. Lung mRNA expression of the C-C chemokine MCP-1 was increased in OVA-indomethacin mice, while there was no difference between the two groups in lung mRNA expression of eotaxin, MIP-1alpha, MIP-1beta, or MIP-2. Histologic examination revealed greater pulmonary interstitial eosinophilia in OVA-indomethacin mice as well. Contrary to our expectations, we conclude that in the BALB/c mouse, cyclooxygenase inhibition during allergen sensitization increases AHR, production of IL-5 and IL-13, and interstitial eosinophilia.

Topics: Animals; Arachidonic Acid; Chemokines, CC; Cyclooxygenase Inhibitors; Female; Hypersensitivity; Immunoglobulin E; Indomethacin; Interleukin-13; Interleukin-4; Interleukin-5; Interleukin-6; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Specific Pathogen-Free Organisms

We investigated the contribution of mast cell chymase in mast cell-dependent angiogenesis using the hamster sponge-implant model, where angiogenesis in the granulation tissue surrounding the subcutaneously implanted sponge was evaluated by measuring the hemoglobin content. Daily local injection of compound 48/80 (3-100 microg/site/day), a potent mast cell activator, induced formation of granulomas and angiogenesis in time- and dose-dependent manners. This angiogenic response was inhibited by chymase inhibitors including chymostatin (> or = 1 nmol/site/day), soybean trypsin inhibitor (SBTI; > or = 1.4 nmol/site/day) and lima bean trypsin inhibitor (LBTI; > or = 3.3 nmol/site/day), but not by a tryptase inhibitor like leupeptin (> or = 700 nmol/site/day). Although pyrilamine (> or = 2,580 nmol/site/day), a histamine H1 receptor antagonist, and protamine (300 microg/site/day) also inhibited angiogenesis, these effects were much less pronounced than those by chymase inhibitors. Furthermore, antigen-induced angiogenesis in hamsters pre-sensitized with ovalbumin was also inhibited by the chymase inhibitors by 60-70%. Our results suggest that chymase is a major mediator in mast cell-mediated angiogenesis.

Topics: Animals; Chymases; Cricetinae; Granuloma; Hemoglobins; Heparin; Histamine; Hypersensitivity; Male; Mast Cells; Mesocricetus; Neovascularization, Pathologic; Oligopeptides; Ovalbumin; p-Methoxy-N-methylphenethylamine; Serine Endopeptidases; Serine Proteinase Inhibitors; Trypsin Inhibitors

A synthetic peptide corresponding to residues 75-84 of HLA-B2702 modulates immune responses in rodents and humans both in vitro and in vivo.. We used a yeast two-hybrid screening, an in vitro biochemical method, and an in vivo animal model.. Two cellular receptors for this novel immunomodulatory peptide were identified using a yeast two-hybrid screen: immunoglobulin binding protein (BiP), a member of the heat shock protein 70 family, and vascular cell adhesion molecule (VCAM)-1. Identification of BiP as a ligand for this peptide confirms earlier biochemical findings, while the interaction with VCAM-1 suggests an alternative mechanism of action. Binding to the B2702 peptide but not to closely related variants was confirmed by ligand Western blot analysis and correlated with immunomodulatory activity of each peptide. In mice, an ovalbumin-induced allergic pulmonary response was blocked by in vivo administration of either the B2702 peptide or anti-VLA-4 antibody.. We propose that the immunomodulatory effect of the B2702 peptide is caused, in part, by binding to VCAM-1, which then prevents the normal interaction of VCAM-1 with VLA-4.

Topics: Amino Acid Sequence; Animals; Carrier Proteins; Endoplasmic Reticulum Chaperone BiP; Female; Heat-Shock Proteins; Histocompatibility Antigens Class I; HSP70 Heat-Shock Proteins; Hypersensitivity; Lung Diseases; Mice; Mice, Inbred BALB C; Molecular Chaperones; Molecular Sequence Data; Ovalbumin; Peptide Fragments; Saccharomyces cerevisiae; Sequence Alignment; Sequence Homology, Amino Acid; Vascular Cell Adhesion Molecule-1

Topics: Animals; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Maternal-Fetal Exchange; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Th1 Cells; Th2 Cells

Tumour necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) have been implicated in the pathogenesis of asthma. The p38 kinase inhibitor, SB 203580 inhibits TNF-alpha and IL-1beta production in vitro and in vivo. In this study the effect of SB 203580 on allergen-induced airway TNF-alpha production and inflammatory cell recruitment was investigated in sensitized Brown Norway rats. The allergen-induced increase in bronchoalveolar lavage (BAL) TNF-alpha was inhibited by SB 203580 at every dose tested (10 - 100 mg kg(-1), p.o.). In contrast, neither ovalbumin-induced eosinophilia or neutrophilia were inhibited by SB 203580 (10 - 100 mg kg(-1), p.o.). In conclusion, SB 203580 inhibits BAL TNF-alpha production by 95% without inhibiting either antigen-induced airway eosinophilia or neutrophilia. This data suggests that either the residual TNF-alpha is sufficient to drive allergen-induced inflammatory cell recruitment into the lung or that TNF-alpha is not involved in allergen-induced inflammatory cell recruitment.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage; Cell Movement; Disease Models, Animal; Enzyme Inhibitors; Female; Hypersensitivity; Imidazoles; Mitogen-Activated Protein Kinases; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Pyridines; Rats; Tumor Necrosis Factor-alpha

Endothelin-1 is a potent bronchoconstrictor peptide with pro-inflammatory and growth-promoting properties. After exposure of sensitized Brown-Norway rats to six repeated ovalbumin exposures, there was an increase in pro-endothelin (ET)-1 messenger RNA compared with saline-exposed control rats 24 h after the final exposure (P < 0.01). ET-1 immunoreactivity was increased sixfold in the bronchial epithelium of the larger conducting airways in the repeated allergen-exposed rats (P < 0.001). After repeated allergen exposure, there were increased rates of DNA synthesis in the airway smooth muscle (ASM) cells (P < 0.001) and epithelial cells (P < 0. 001) compared with saline-exposed controls, as measured by bromodeoxyuridine incorporation. Treatment with a dual endothelin A and B (ET(A+B)) receptor antagonist caused a significant attenuation in both ASM (P < 0.001) and epithelial cell (P < 0.001) bromodeoxyuridine incorporation compared with the allergen-challenged and vehicle-treated group. The dual ET(A+B) antagonist attenuated eosinophil recruitment into the airways (P < 0. 05) but had no significant effect on increased bronchial reactivity to acetylcholine in allergen-exposed rats. Increased levels of ET-1 in the airways may contribute to inflammation and ASM and epithelial cell DNA synthesis after repeated allergen exposure. Such processes may underlie increased proliferation of resident cells leading to airway wall remodeling in asthmatics.

Topics: Allergens; Animals; DNA; Endothelin-1; Epithelial Cells; Hypersensitivity; Muscle, Smooth; Ovalbumin; Rats; Rats, Inbred BN; Respiratory Physiological Phenomena; Up-Regulation

The overall purpose of this study was to investigate how airborne house dust particles may contribute to an allergic immune response, and thereby also to asthma and other respiratory symptoms. The following aims were set: first, to quantify and characterize indoor suspended particulate matter (SPM) with regard to amount, as well as elemental and size distribution, second, to identify possible mechanisms by which SPM may affect the allergic immune response. A vast majority of the particles in SPM samples from homes in Oslo were found to be less than 2.5 microm in diameter. This PM(2.5) fraction contained, in addition to a large amount of sulfur aerosols and silicates, a lot of soot particles. Most of these were less than 1 microm in diameter. Using an immunogold labeling technique, we found that these soot particles carried cat, dog and birch allergens on their surface. These results show that indoor SPM contains a lot of potential allergen carriers, i.e. soot particles (carbon aggregates), most of them less that 1 microm in diameter and therefore able to transport allergens deep into the respiratory tree. We further found that diesel exhaust particles (DEP), which is likely the main soot component of SPM, adsorbed several well-known allergens in vitro. Furthermore, SPM was found to elicit a local lymph node inflammatory response, and to have an adjuvant activity on the production of IgE antibodies to ovalbumin (OA).

Topics: Adjuvants, Immunologic; Air Pollution, Indoor; Allergens; Animals; Dust; Female; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin

Allergic asthma, which is present in as many as 10% of individuals in industrialized nations, is characterized by chronic airway inflammation and hyperreactivity induced by allergen-specific Th2 cells secreting interleukin-4 (IL-4) and IL-5. Because Th1 cells antagonize Th2 cell functions, it has been proposed that immune deviation toward Th1 can protect against asthma and allergies. Using an adoptive transfer system, we assessed the roles of Th1, Th2, and Th0 cells in a mouse model of asthma and examined the capacity of Th1 cells to counterbalance the proasthmatic effects of Th2 cells. Th1, Th2, and Th0 lines were generated from ovalbumin (OVA)-specific T-cell receptor (TCR) transgenic mice and transferred into lymphocyte-deficient, OVA-treated severe combined immunodeficiency (SCID) mice. OVA-specific Th2 and Th0 cells induced significant airway hyperreactivity and inflammation. Surprisingly, Th1 cells did not attenuate Th2 cell-induced airway hyperreactivity and inflammation in either SCID mice or in OVA-immunized immunocompetent BALB/c mice, but rather caused severe airway inflammation. These results indicate that antigen-specific Th1 cells may not protect or prevent Th2-mediated allergic disease, but rather may cause acute lung pathology. These findings have significant implications with regard to current therapeutic goals in asthma and allergy and suggest that conversion of Th2-dominated allergic inflammatory responses into Th1-dominated responses may lead to further problems.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Cytokines; Disease Models, Animal; Eosinophils; Histocytochemistry; Hypersensitivity; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Mice, SCID; Mice, Transgenic; Ovalbumin; Th1 Cells; Th2 Cells

Inflammatory-cell infiltration and epithelial modifications are prominent lesions of the bronchial mucosa in asthma and in experimental allergic bronchopulmonary inflammation. However, the recruitment of inflammatory cells and their relationship to the epithelial modifications and to functional alterations such as bronchopulmonary hyperreactivity (BHR) are less known. We studied the mechanisms of antigen-dependent inflammatory-cell recruitment to the lungs and the associated lesions and their relationship using drug- and antibody-dependent cell-depletion procedures. A single intranasal ovalbumin challenge in BP2 mice was found to induce hyperreactivity within 1 h after challenge, followed by the massive infiltration of immunoglobulin (Ig)E-bearing polymorphonuclear leukocytes (PMN), and eosinophils, and by a mucous-cell metaplasia of the bronchiolar epithelium. Similarly challenged BALB/c mice did not exhibit BHR, despite a moderate recruitment of inflammatory cells and mucous-cell metaplasia. Inflammatory-cell recruitment, mucous-cell metaplasia, and BHR were prevented by prior antibody-dependent depletion of CD3(+) lymphocytes and partially inhibited by the depletion of CD4(+) lymphocytes. Treatment with the granulocytopenic drug vinblastine before challenge completely abolished the recruitment of granulocytes without affecting the antigen-induced mucous-cell metaplasia. In this study two new key elements of the murine model of allergic pulmonary inflammation are described: the recruitment of IgE-bearing PMN between 3 and 72 h after challenge, and the dissociation between granulocytes and mucous-cell metaplasia.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Lymphocytes; Mast Cells; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin

Allergic sensitization in asthma develops as a consequence of complex interactions between T cells and antigen-presenting cells. We have developed several in vivo models to study allergen-specific T cell and B cell function and their relevance to allergic airway hyperresponsiveness (AHR), focusing on the role of the costimulatory molecules CD80 and CD86. Treatment of mice with anti-CD86, but not anti-CD80, significantly inhibited increased serum levels of ovalbumin (OA)-specific IgE and IgG1, airway eosinophilia, and AHR both after 10 d of OA aerosol exposure (in the absence of adjuvant) and after intraperitoneal sensitization followed by repeated airway challenges. Inhibition of AHR was associated with decreased IL-4 and IL-5 levels in the BAL fluid of sensitized mice, suggesting impaired Th2 function in anti-CD86-treated animals. This effect was not seen when mice received treatment only before allergen challenge, indicating that anti-CD86 acts through inhibition of allergic sensitization and not simply by inhibiting the influx of inflammatory cells. These data suggest that the CD86 costimulatory ligand plays a major role in the development of allergic inflammation and AHR in allergen-challenged mice. Further, this study demonstrates that T-B cell interactions during allergic sensitization are amenable to therapeutic manipulation and that selective blockade of accessory signals can be an effective means for modulating distinct T cell functions.

Topics: Animals; Antibodies; Antibody Formation; Antigens, CD; B7-2 Antigen; Bronchoalveolar Lavage Fluid; Cytokines; Electric Stimulation; Eosinophilia; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lung; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Muscle, Smooth; Ovalbumin; Respiratory Hypersensitivity; Trachea

It is widely accepted that immunoglobulin (Ig)E triggers immediate hypersensitivity responses by activating a cognate high-affinity receptor, FcepsilonRI, leading to mast cell degranulation with release of vasoactive and proinflammatory mediators. This apparent specificity, however, is complicated by the ability of IgE to bind with low affinity to Fc receptors for IgG, FcgammaRII and III. We have addressed the in vivo significance of this interaction by studying IgE-mediated passive systemic anaphylaxis in FcgammaR-deficient mice. Mice deficient in the inhibitory receptor for IgG, FcgammaRIIB, display enhanced IgE-mediated anaphylactic responses, whereas mice deficient in an IgG activation receptor, FcgammaRIII, display a corresponding attenuation of IgE-mediated responses. Thus, in addition to modulating IgG-triggered hypersensitivity responses, FcgammaRII and III on mast cells are potent regulators of IgE-mediated responses and reveal the existence of a regulatory pathway for IgE triggering of effector cells through IgG Fc receptors that could contribute to the etiology of the atopic response.

Topics: Anaphylaxis; Animals; Antigens, CD; Body Temperature; Bone Marrow Cells; Histocytochemistry; Hypersensitivity; Ileum; Immunoglobulin E; Immunoglobulin G; Mast Cells; Mice; Mice, Knockout; Ovalbumin; Receptors, IgG

A recent increase in allergic disorders has coincided with a decrease in infections, including tuberculosis. Although an inverse association between tuberculin responses and atopic disorders was reported, it was not known how T-helper (Th)1-biased immune responses to Mycobacterium tuberculosis influenced Th2-dominant responses to allergens. We examined whether M. tuberculosis could modulate ovalbumin (OVA)-induced eosinophilic inflammation in the murine trachea in a manner that transcended the barrier of antigen specificity. We found that CD4(+) T cells primed with OVA in complete Freund's adjuvant (CFA) inhibited OVA-induced tracheal eosinophilia through interferon (IFN)-gamma secretion. Immunization with an irrelevant antigen in CFA or with OVA in incomplete Freund's adjuvant failed to induce suppressor cells. In vitro experiments confirmed that both M. tuberculosis and OVA (as opposed to either one alone) were necessary to evoke polarized development toward a Th1-like phenotype through interleukin-12 secretion. These results indicate that exposure to an allergen along with M. tuberculosis switches development of allergen-specific T cells toward a Th1 phenotype, which, in turn, downregulates allergic manifestations in an antigen-specific manner. The possible implications of these results are discussed in the context of the causal relationship between a decrease in tuberculosis and an increase in allergic disorders.

Topics: Animals; Asthma; CD4-Positive T-Lymphocytes; Dose-Response Relationship, Drug; Eosinophilia; Hypersensitivity; Interferon-gamma; Interleukin-12; Interleukin-4; Mice; Mice, Inbred BALB C; Mice, Transgenic; Models, Biological; Mycobacterium tuberculosis; Ovalbumin; Phenotype; Th1 Cells; Th2 Cells; Tracheal Diseases

T lymphocytes have a central regulatory role in the pathogenesis of asthma. We delineated the participation of lymphocytes in the acute allergic and chronic tolerant stages of a murine model of asthma by characterizing the various subsets of lymphocytes in bronchoalveolar lavage and lung tissue associated with these responses. Acute (10-day) aerosol challenge of immunized C57BL/6J mice with ovalbumin resulted in airway eosinophilia, histological evidence of peribronchial and perivascular airway inflammation, clusters of B cells and TCRgammadelta cells in lung tissue, increased serum IgE levels, and airway hyperresponsiveness to methacholine. In mice subjected to chronic (6-week) aerosol challenge with ovalbumin, airway inflammation and serum IgE levels were significantly attenuated and airway hyperresponsiveness was absent. The marked increases in lung B and T cell populations seen in the acute stage were also significantly reduced in the chronic stage of this model. Thus, acute ovalbumin challenge resulted in airway sensitization characteristic of asthma, whereas chronic ovalbumin challenge elicited a suppressed or tolerant state. The transition from antigenic sensitization to tolerance was accompanied by shifts in lymphocyte profiles in the lung and bronchoalveolar lavage fluid.

Topics: Airway Resistance; Animals; Asthma; B-Lymphocytes; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Fluorescent Antibody Technique; Hypersensitivity; Immunoglobulin E; Lymphocytes; Male; Methacholine Chloride; Mice; Mice, Inbred C57BL; Ovalbumin; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocytes; Time Factors

Cell accumulation and CC chemokine production were assessed in the peritoneal cavity of ovalbumin (OVA)-sensitized mice following antigen challenge. Intraperitoneal challenge with OVA induced a significant eosinophil influx from 6 h post-challenge with increased numbers persisting at 24 h. At 6 h there was also a marked presence of neutrophils. Messenger RNA expression and protein levels for the chemokines RANTES and MIP-1 alpha were measured in the cell pellets and supernatants, respectively, from peritoneal washes following OVA challenge. RANTES mRNA was detected from 2 h to 4 h following OVA injection, whereas mRNA for MIP-1 alpha was only detectable at 4 h. RANTES protein was first detected from 4 h after OVA injection and by 24 h the protein levels had increased further. Basal levels of MIP-1 alpha were detected in peritoneal washes. These levels peaked at 2 h after OVA challenge and rapidly declined to basal levels by 6 h. A functional role for the chemokines was assessed using neutralizing polyclonal antibodies. Co-injection of OVA with anti-RANTES antibodies resulted in a significant inhibition of eosinophil infiltration into the cavity at 6 h and 24 h (63% and 52% inhibition, respectively) without significantly influencing the number of neutrophils present. In contrast, injection of anti-MIP-1 alpha antibodies only inhibited neutrophil migration at the 6 h time point by 44% without significantly affecting the accumulation of eosinophils. These results demonstrate an important role for RANTES in mediating eosinophil influx in allergic inflammation and a contrasting role for MIP-1 alpha in mediating neutrophil recruitment.

Topics: Animals; Cell Movement; Chemokine CCL4; Chemokine CCL5; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immune Sera; Injections, Intraperitoneal; Macrophage Inflammatory Proteins; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Peritoneal Cavity; Peritonitis; Time Factors

A recently advanced hypothesis suggests that decreased exposure to T-helper (Th) 1-inducing agents causes Th2-biased differentiation in response to concomitant allergens. We therefore examined the effect of pre-immunization with killed Mycobacterium bovis and killed M. vaccae which are known to be very potent inducers of Thl immune response, on serum IgE response in ovalbumin (OVA)-sensitized newborn mice. Eighty-four newborn Balb/c mice were divided into four groups and were immunized intraperitoneally 24 h after birth with 50 microl of 5 x 10(4) colony-forming units (c.f.u.) of killed M. bovis in group I (M. bovis group, n = 19), with 25 microl of 2.5 x 10(8) c.f.u. of killed M. vaccae plus 25 microl of 5 x 10(4) c.f.u. of killed M. bovis in group II (M. vaccae + M. bovis group, n = 28) and with 50 microl of only phosphate-buffered saline (PBS) in group III (no mycobacterial immunization, n = 18). No injection was applied to mice in group IV (control group, n = 19). Starting from 8 weeks of age, all mice except the control group were sensitized with 0.5 ml of 20 mg/ml OVA administered intraperitoneally 7 times every other day. Thirty days after the final injection, all animals except those in the control group were challenged with an aerosol of 2 mg/ml OVA. Forty-eight hours later, blood was collected from all mice for determination of serum IgE levels. A statistically significant difference was observed in the serum total IgE levels between groups III and IV (p = 0.0099), indicating that the mice were successfully sensitized with OVA. Serum total IgE values of the female mice in M. bovis group were found to be significantly lower than group III (p = 0.009), while no difference was observed in males. Serum total IgE levels of the M. vaccae + M. bovis group were found to be significantly lower than group III both in male and female mice (p < 0.0001 and p = 0.0001, respectively). Female values were even lower than controls (p = 0.0092). Pre-immunization in the newborn period with killed M. bovis alone or in addition to M. vaccae may potentially be helpful in down-regulating an IgE response.

Topics: Animals; Bacterial Vaccines; Female; Hypersensitivity; Immunization; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Mycobacterium; Mycobacterium bovis; Ovalbumin; Pregnancy; Statistics, Nonparametric; Vaccines, Inactivated

Recent reports have demonstrated that feeding small amounts of antigen conjugated to the B subunit of cholera toxin (CTB) suppress immune responses in experimental models of certain T(h)1-based autoimmune diseases. We have established a model of aerosol sensitization leading to T(h)2-mediated allergic immune responses in BALB/c mice. In the present study two different antigens, the dietary antigen ovalbumin (OVA) and the inhalant allergen Bet v 1 (the major birch pollen allergen), chemically coupled to recombinant CTB were tested for their potential to influence T(h)2-like immune responses. Intranasal administration of OVA-CTB prior to sensitization with OVA led to a significant decrease of antigen-specific IgE antibody levels, but a marked increase of OVA-specific IgG2a antibodies as compared to non-pretreated, sensitized animals. Antigen-specific lympho-proliferative responses in vitro were reduced by 65% in the pretreated group; IL-5 and IL-4 production were decreased in responder cells of lungs and spleens of nasally pretreated mice. In contrast, mucosal administration of rBet v 1-CTB conjugates prior to sensitization led to an up-regulation of allergen-specific IgE, IgG1 and IgG2a, increased in vitro lympho-proliferative responses as well as augmented production of IL-5, IL-4, IL-10 and IFN-gamma. Intranasal administration prior to sensitization of unconjugated allergens showed also contrasting effects: OVA could not significantly influence antigen-specific antibody or cytokine production, whereas intranasal pretreatment with unconjugated Bet v 1 suppressed allergen-specific immune responses in vivo and in vitro. These results demonstrated that the two antigens-in conjugated as in unconjugated form-had different effects on the T(h)2 immune responses. We therefore conclude that the tolerogenic or immunogenic properties of CTB-and probably also other antigen-delivery systems-strongly depend on the nature of the coupled antigen-allergen.

Topics: Administration, Intranasal; Allergens; Animals; Cholera Toxin; Disease Models, Animal; Female; Hypersensitivity; Immune Tolerance; Immunity, Mucosal; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukins; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Proteins; Spleen; Toxoids

The effects of lactic dehydrogenase virus (LDV) infection on allergic eosinophil reaction and IL-5 gene expression were studied. LDV infection suppressed antigen-induced eosinophil recruitment into the peritoneal cavity in sensitized mice. The elevation of IL-5 gene expression in the spleen and mesenteric lymph nodes 6 h after ovalbumin challenge was significantly suppressed in LDV-infected mice compared with uninfected (control) mice. The expression of the interferon-gamma and IL-2 genes in the spleen, but not in mesenteric lymph nodes, was significantly suppressed in LDV-infected mice compared with control mice. The present results suggest, that suppression of IL-5 gene expression by LDV infection may not be mediated by a mutual inhibitory mechanism between Th1 and Th2 cells.

Topics: Animals; Antigens; Arterivirus Infections; Base Sequence; DNA Primers; Eosinophils; Gene Expression; Hypersensitivity; Interferon-gamma; Interleukin-2; Interleukin-5; Lactate dehydrogenase-elevating virus; Lymph Nodes; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Peritoneal Cavity; Spleen; Th1 Cells; Th2 Cells

CD4 T helper (Th) type 1 and Th2 cells have been identified in the airways of asthmatic patients. Th2 cells are believed to contribute to pathogenesis of the disease, but the role of Th1 cells is not well defined. In a mouse model, we previously reported that transferred T cell receptor-transgenic Th2 cells activated in the respiratory tract led to airway inflammation with many of the pathologic features of asthma, including airway eosinophilia and mucus production. Th1 cells caused inflammation with none of the pathology associated with asthma. In this report, we investigate the role of Th1 cells in regulating airway inflammation. When Th1 and Th2 cells are transferred together into recipient mice, there is a marked reduction in airway eosinophilia and mucus staining. To address the precise role of Th1 cells, we asked (i), Are Th2-induced responses inhibited by interferon (IFN)-gamma? and (ii) Can Th1 cells induce eosinophilia and mucus in the absence of IFN-gamma? In IFN-gamma receptor(-/-) recipient mice exposed to inhaled antigen, the inhibitory effects of Th1 cells on both airway eosinophilia and mucus production were abolished. In the absence of IFN-gamma receptor signaling, Th1 cells induced mucus but not eosinophilia. Thus, we have identified new regulatory pathways for mucus production; mucus can be induced by Th2 and non-Th2 inflammatory responses in the lung, both of which are inhibited by IFN-gamma. The blockade of eosinophilia and mucus production by IFN-gamma likely occurs through different inhibitory pathways that are activated downstream of Th2 cytokine secretion and require IFN-gamma signaling in tissue of recipient mice.

Topics: Animals; Asthma; Disease Models, Animal; Eosinophilia; Flow Cytometry; Hypersensitivity; Inflammation; Interferon-gamma; Interleukins; Lung; Mice; Mice, Knockout; Mucus; Ovalbumin; Receptors, Interferon; T-Lymphocytes, Helper-Inducer; Th1 Cells; Th2 Cells

Because bradykinin has potent inflammatory actions, this molecule may be involved in the late allergic response (LAR). We investigated the role of the molecule in airway microvascular hyperpermeability during the LAR. Three weeks after ovalbumin (OVA) sensitization, animals were pretreated with bradykinin B2 receptor antagonist HOE 140 or vehicle for 30 min before the OVA inhalation challenge. The occurrence of LAR was judged by a two-fold increase in transpulmonary pressure (Ptp) from the baseline values. The microvascular permeability in the trachea was assessed by an index defined as the ratio of the area of vasculature labeled by the Monastral blue dye (area density percent). Significant microvascular hyperpermeability were observed during the LAR. The bradykinin concentrations in the bronchoalveolar lavage-fluid (BAL-f) were increased during the LAR. HOE 140 (0.1-10 mg/kg, s.c.) inhibited the airway microvascular hyperpermeability during the LAR dose-dependently. These findings suggest that bradykinin may play an important role in microvascular hyperpermeability during the LAR.

Topics: Animals; Asthma; Bradykinin; Bradykinin Receptor Antagonists; Bronchoalveolar Lavage Fluid; Capillary Permeability; Guinea Pigs; Hypersensitivity; Male; Microcirculation; Ovalbumin; Receptor, Bradykinin B2; Trachea

Airway inflammation plays a major role in human asthma. Increasing evidence points to a close correlation between eosinophil infiltration and allergic lung disease. A new murine model of eosinophilic lung inflammation has recently been developed; it consists of immunizing mice with small fragments of solidified hen egg white implanted (EWI) into the subcutaneous tissue. In this model, which is further characterized here, mice challenged with ovalbumin (OVA) present an intense and persistent lung eosinophilia, as well as histopathological findings that resemble human asthma. In the present work, the effect of oral tolerance on the development of allergic lung inflammation in B6 mice immunized with antigen plus adjuvant or with EWI is investigated. It was found that in mice rendered orally tolerant by previous exposure to antigen in the drinking water, the T-helper type 2 cell (Th2)-associated allergic responses in both protocols of immunization were almost completely abolished. The allergic responses were assessed by pulmonary and bone marrow eosinophilia, lung histopathology and antigen-specific IgE and IgG1 production. These findings provide the first indication that Th2-associated lung pathology can be prevented by oral tolerance.

Topics: Administration, Oral; Adsorption; Aluminum Hydroxide; Animals; Egg White; Eosinophil Peroxidase; Eosinophils; Female; Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin Isotypes; Immunotherapy; Mice; Mice, Inbred C57BL; Ovalbumin; Peroxidases; Pulmonary Eosinophilia; Th2 Cells

Signal transducers and activators of transcription 6 (STAT6) is essential for interleukin 4-mediated responses, including class switching to IgE and induction of type 2 T helper cells. To investigate the role of STAT6 in allergic asthma in vivo, we developed a murine model of allergen-induced airway inflammation. Repeated exposure of actively immunized C57BL/6 mice to ovalbumin (OVA) aerosol increased the level of serum IgE, the number of eosinophils in bronchoalveolar lavage (BAL) fluid, and airway reactivity. Histological analysis revealed peribronchial inflammation with pulmonary eosinophilia in OVA-treated mice. In STAT6-deficient (STAT6-/-) C57BL/6 mice treated in the same fashion, there were no eosinophilia in BAL and significantly less peribronchial inflammation than in wild-type mice. Moreover STAT6-/- mice had much less airway reactivity than wild-type mice. These findings suggest that STAT6 plays a crucial role in the pathogenesis of allergen-induced airway inflammation.

Topics: Animals; Bronchial Hyperreactivity; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunoglobulin E; Immunoglobulin M; Inflammation; Lung; Mice; Mice, Inbred Strains; Mice, Knockout; Ovalbumin; Signal Transduction; STAT6 Transcription Factor; Trans-Activators

This study describes the action of the sesquiterpene polygodial, the major constituent isolated from the bark of Drymis winteri in the guinea pig ileum and trachea in vitro. Polygodial (5 to 128 microM), added for 20 min, did not affect the resting tone of the preparations, but caused graded inhibition, associated in some cases with rightward displacement of the acetylcholine, histamine (1 nM to 10 microM), bradykinin (0.1 nM to 1 microM) and KCl (1 to 100 mM)-contraction response curves. When assessed in the guinea-pig trachea, polygodial (5 to 342 microM) caused significant inhibition of bradykinin (10 pM to 1 microM), 9,11-dideoxy-9alpha,11alpha-methano-epoxy prostaglandin F2alpha (0.1 to 1000 nM) and KCl (1 to 100 mM)-induced contractions, although the action against bradykinin was not concentration-dependent. Polygodial (5 to 80 microM) caused a small but significant shift to the right of substance P and also the selective agonist of tachykinin NK2 receptor [beta-Ala8]neurokinin A-(4-10)-induced contractions in guinea pig trachea. This action of polygodial seems to be quite selective towards tachykinin NK2 receptors since up to 432 microM, polygodial had no effect against contraction caused by tachykinin NK1 receptor agonist, substance P methyl ester. When tested in the guinea-pig trachea from animals which had been actively sensitised to ovalbumin, polygodial (30 to 40 microM) caused time and concentration-dependent inhibition of ovalbumin-mediated contraction. In addition, polygodial (85 to 342 microM) inhibited contraction induced by compound 48/80 (1 to 1000 microg/ml), in the guinea-pig trachea from non-sensitised animals. These findings and those from our previous study are consistent with the notion that the main sesquiterpene polygodial isolated from the bark of D. winteri is responsible for most, if not all, of the relevant pharmacological action reported previously for the extract of this plant. Thus, polygodial could be of potential value in the development of a new drug for the treatment of asthma, allergy and other inflammatory processes.

Topics: Animals; Female; Guinea Pigs; Histamine; Hypersensitivity; Ileum; In Vitro Techniques; Male; Muscle Relaxation; Muscle, Smooth; Ovalbumin; Plants, Medicinal; Sesquiterpenes; Trachea

Exposure of sensitized individuals to antigen can induce allergic responses in the respiratory tract, manifested by early and late phases of vasodilatation, plasma leakage, leukocyte influx, and bronchoconstriction. Similar responses can occur in the skin, eye, and gastrointestinal tract. The early-phase response involves mast cell mediators and the late-phase response is leukocyte dependent, but the mechanism of leakage is not understood. We sought to identify the leaky blood vessels, to determine whether these vessels contained endothelial gaps, and to analyze the relationship of the gaps to adherent leukocytes, using biotinylated lectins or silver nitrate to stain the cells in situ and Monastral blue as a tracer to quantify plasma leakage. Most of the leakage occurred in postcapillary venules (< 40-microns diameter), whereas most of the leukocyte migration (predominantly neutrophils) occurred in collecting venules. Capillaries and arterioles did not leak. Endothelial gaps were found in the leaky venules, both by silver nitrate staining and by scanning electron microscopy, and 94% of the gaps were distinct from sites of leukocyte adhesion or migration. We conclude that endothelial gaps contribute to both early and late phases of plasma leakage induced by antigen, but most leakage occurs upstream to sites of leukocyte adhesion.

Topics: Allergens; Animals; Capillary Permeability; Cell Movement; Endothelium, Vascular; Hypersensitivity; Indicators and Reagents; Indoles; Leukocytes; Male; Organometallic Compounds; Ovalbumin; Rats; Rats, Inbred F344; Rats, Inbred Lew; Rats, Wistar; Silver Staining; Time Factors; Trachea

The effect of chemical denaturation of ovalbumin (OVA) on the induction of oral tolerance of reaginic antibody responses was studied. Both urea-denatured OVA (UD-OVA) and carboxymethylated UD-OVA (CM-OVA) were purified by centrifugation. When compared with OVA and UD-OVA, CM-OVA had the least sensitizing capacity and allergenicity in IgE responses to OVA. BALB/c IgE, IgG1 and IgG antibody responses were suppressed by OVA, but not by UD-OVA or CM-OVA, fed prior to sensitization with OVA, UD-OVA, or CM-OVA in alum, respectively. The priming effect of specific IgG and IgG1 antibody responses was induced by CM-OVA fed prior to sensitization with OVA or CM-OVA. The proliferation of BALB/c spleen cells and their secretion of T helper type 2 (Th2) cytokines interleukin-4 (IL-4) and IL-5 were also orally tolerized by OVA, but not by denatured OVA. Although denatured OVA is hypoallergenic, the present result indicates that denaturation of a soluble protein prevents the induction of oral tolerance of Th2 responses.

Topics: Administration, Oral; Animals; Antibody Formation; Cell Division; Cytokines; Female; Hypersensitivity; Immune Tolerance; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Protein Denaturation; Rats; Rats, Sprague-Dawley; Reagins; Spleen; Th2 Cells

Allergic disorders are mediated by T lymphocytes secreting T helper 2 (Th2) cytokines, interleukin-4 (IL-4) and interleukin-5 (IL-5), resulting in high levels of serum immunoglobulin E (IgE) and recruitment of eosinophils. One of the treatment strategies is to downregulate the Th2 component by inducing a T helper 1 (Th1) response to the relevant allergen, because Th1 and Th2 cytokines are thought to be mutually antagonistic. In this study, we examined the effects of Mycobacterium vaccae, a potent inducer of Th1 immunity, on allergic responses in a murine model. A single injection of M. vaccae into ovalbumin (OVA)-preimmunized BALB/c mice suppressed serum IgE over a wide dose range (10(7), 10(8) or 10(9) M. vaccae). Further experiments, using 10(7) M. vaccae injected twice, showed that this treatment inhibited not only serum IgE, but also the potential for ovalbumin-induced IL-5 production by spleen cells. This non-specific ability of a mycobacterium to decrease Th2 activity, even when not presented together with the allergen, is in agreement with recent epidemiological studies on the impact of bacillus Calmette-Guérin (BCG) vaccination, and of other potent Th1 stimuli, on the incidence of atopy. The suppression of serum IgE and allergen-specific IL-5 synthesis by M. vaccae suggest that this organism is likely to have clinical application in the immunotherapy of allergy.

Topics: Animals; Cells, Cultured; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Hypersensitivity; Immunoglobulin E; Interferon-gamma; Interleukin-2; Interleukin-5; Mice; Mice, Inbred BALB C; Mycobacterium; Ovalbumin; Spleen; Th1 Cells; Th2 Cells; Vaccines, Inactivated

1. We investigated the inhibitory effects of the cysteinyl leukotriene (CysLT1) receptor antagonists, pranlukast and zafirlukast, on 35SO4 labelled mucus output, in vitro, in guinea-pig trachea, induced by leukotriene D4 (LTD4) or by antigen challenge of sensitized animals. Agonists and antagonists were administered mucosally, except in selected comparative experiments where drugs were administered both mucosally and serosally to assess the influence of the epithelium on evoked-secretion. 2. LTD4 increased 35SO4 output in a concentration-related manner with a maximal increase of 23 fold above controls at 100 microM and an approximate EC50 of 2 microM. Combined mucosal and serosal addition of LTD4 did not significantly affect the secretory response compared with mucosal addition alone. Neither LTC4 nor LTE4 (10 microM each) affected 35SO4 output. Pranlukast or zafirlukast significantly inhibited 10 microM LTD4-evoked 35SO4 output in a concentration-dependent fashion, with maximal inhibitions of 83% at 10 microM pranlukast and 78% at 10 microM zafirlukast, and IC50 values of 0.3 microM for pranlukast and 0.6 microM for zafirlukast. Combined mucosal and serosal administration of the antagonists (5 microM each) gave degrees of inhibition of mucosal-serosal 10 microM LTD4-evoked 35SO4 output similar to those of the drugs given mucosally. Pranlukast (0.5 microM) caused a parallel rightward shift of the LTD4 concentration-response curve with a pKB of 7. Pranlukast did not inhibit ATP-induced 35SO4 output. 3. Ovalbumin (10-500 microg ml(-1) challenge of tracheae from guinea-pigs actively sensitized with ovalbumin caused a concentration-related increase in 35SO4 output with a maximal increase of 20 fold above vehicle controls at 200 microg ml(-1). The combination of the antihistamines pyrilamine and cimetidine (0.1 mM each) did not inhibit ovalbumin-induced 35SO4 output in sensitized guinea-pigs. Neither mucosal (10 microM or 100 microM) nor mucosal-serosal (100 microM) histamine had any significant effect on 35SO4 output. 4. Pranlukast or zafirlukast (5 microM each) significantly suppressed ovalbumin-induced secretion in tracheae from sensitized guinea-pigs by 70% and 65%, respectively. 5 We conclude that LTD4 or ovalbumin challenge of sensitized animals provokes mucus secretion from guinea-pig trachea in vitro and this effect is inhibited by the CysLT1 receptor antagonists pranlukast and zafirlukast. These antagonists may be beneficial in the treatment of allerg

Topics: Adenosine Triphosphate; Animals; Anti-Asthmatic Agents; Binding, Competitive; Chromones; Cysteine; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Indoles; Leukotriene Antagonists; Leukotriene D4; Male; Membrane Proteins; Mucus; Ovalbumin; Passive Cutaneous Anaphylaxis; Phenylcarbamates; Receptors, Leukotriene; Sulfates; Sulfonamides; Sulfur Radioisotopes; Tosyl Compounds; Trachea

The complex pathophysiology of lung allergic inflammation and bronchial hyperresponsiveness (BHR) that characterize asthma is achieved by the regulated accumulation and activation of different leukocyte subsets in the lung. The development and maintenance of these processes correlate with the coordinated production of chemokines. Here, we have assessed the role that different chemokines play in lung allergic inflammation and BHR by blocking their activities in vivo. Our results show that blockage of each one of these chemokines reduces both lung leukocyte infiltration and BHR in a substantially different way. Thus, eotaxin neutralization reduces specifically BHR and lung eosinophilia transiently after each antigen exposure. Monocyte chemoattractant protein (MCP)-5 neutralization abolishes BHR not by affecting the accumulation of inflammatory leukocytes in the airways, but rather by altering the trafficking of the eosinophils and other leukocytes through the lung interstitium. Neutralization of RANTES (regulated upon activation, normal T cell expressed and secreted) receptor(s) with a receptor antagonist decreases significantly lymphocyte and eosinophil infiltration as well as mRNA expression of eotaxin and RANTES. In contrast, neutralization of one of the ligands for RANTES receptors, macrophage-inflammatory protein 1alpha, reduces only slightly lung eosinophilia and BHR. Finally, MCP-1 neutralization diminishes drastically BHR and inflammation, and this correlates with a pronounced decrease in monocyte- and lymphocyte-derived inflammatory mediators. These results suggest that different chemokines activate different cellular and molecular pathways that in a coordinated fashion contribute to the complex pathophysiology of asthma, and that their individual blockage results in intervention at different levels of these processes.

Topics: Animals; Antibodies; Asthma; Chemokine CCL11; Chemokine CCL4; Chemokine CCL5; Chemokines, CC; Chemotactic Factors, Eosinophil; Cytokines; Disease Models, Animal; Hypersensitivity; Immunohistochemistry; Inflammation; Leukocytes; Lung; Macrophage Inflammatory Proteins; Mice; Mice, Inbred Strains; Monocyte Chemoattractant Proteins; Ovalbumin; RNA, Messenger

Systemic administration of IL-12 can prevent airway hyperresponsiveness (AHR) in mice after sensitization and repeated allergen challenge. However, systemic IL-12 has been associated with severe adverse effects.. We determined whether IL-12 administration to the airways in a dose sufficiently low so as not to result in systemic effects can modify allergic inflammation and AHR after allergen challenge.. Mice were sensitized to ovalbumin by intraperitoneal injection and challenged with ovalbumin aerosol on 3 consecutive days. During the period of challenge, IL-12 was administered intranasally following 2 regimens, designated high (1500 ng) or low (150 ng). We monitored airway responsiveness to inhaled methacholine by barometric body plethysmography, lung inflammatory cells, local cytokine production, and, to assess systemic effects of IL-12 treatment, spleen weights and numbers of eosinophils in the bone marrow.. Allergen challenge resulted in increases in airway responsiveness and in numbers of lung eosinophils. These increases were prevented by both high- and low-dose IL-12. Additionally, IL-12 administration resulted in enhanced local interferon-gamma production and prevented the increases in local IL-4 and IL-5 production after airway challenge. A high dose, but not a low dose, of IL-12 resulted in increased spleen weights and prevented the increase in numbers of bone marrow eosinophils after allergen challenge.. These data indicate that local administration of IL-12 can prevent AHR and reduce lung eosinophilia after allergen challenge in sensitized mice without eliciting systemic adverse effects. IL-12 exerts these effects by inducing local T(H1)-type responses in the airways in a setting that is normally dominated by T(H2)-type responses.

Topics: Allergens; Animals; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Eosinophilia; Female; Hypersensitivity; Immunoglobulin G; Interferon-gamma; Interleukin-12; Interleukin-4; Interleukin-5; Leukocytes, Mononuclear; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Proteins

1. It has been shown that activation of protein tyrosine kinases is the earliest detectable signalling response to FcepsilonRI cross-linking on mast cell. Following tyrosine kinase activation, a family of mitogen-activated protein kinases (MAPKs) was found to be activated as well. The present study examined the role of MAPK signalling cascade in in vitro model of allergic asthma using a specific MAPK kinase inhibitor PD 098059. 2. Guinea-pigs were passively sensitized with IgG antibody raised against ovalbumin (OA). Effects of PD 098059 on OA-induced anaphylactic contraction of isolated bronchi and release of histamine and peptidoleukotrienes from chopped lung preparations were studied. 3. PD 098059 (10-50 microM) produced only minor reduction of maximal OA-induced bronchial contraction. In contrast, the rate of relaxation of OA-induced bronchial contraction was markedly faster in the presence of PD 098059 than the vehicle control in a concentration-dependent manner. 4. These observations corroborate well with the inability of PD 098059 (5-50 microM) to substantially block the OA-induced release of histamine and with marked inhibition of OA-induced release of peptidoleukotrienes from lung fragments in the presence of PD 098059. Exogenous arachidonic acid-induced release of peptidoleukotrienes from lung fragments was not blocked by PD 098059. 5. In immunoblotting study, we found that p42MAPK was constitutively expressed in guinea-pig bronchi. However, treatment with OA, histamine or LTD4 did not cause activation of p42MAPK. These findings together with the lack of inhibitory effects of PD 098059 on bronchial contraction induced by histamine or LTD4 suggest that histamine- and LTD4-induced bronchial contractions are not mediated by p42MAPK activation. 6. Taken together, our findings show that inhibition of MAPK signalling cascade by PD 098059 significantly reduced the OA-triggered release of peptidoleukotrienes leading to rapid relaxation of anaphylactic bronchial contraction. On the other hand, p42MAPK did not play a role in histamine- or LTD4-induced bronchial smooth muscle contraction suggesting that PD 098059 exerts its inhibitory effects on OA-induced bronchial contraction primarily through inhibition of peptidoleukotrienes release from mast cells.

Topics: Analysis of Variance; Animals; Asthma; Bronchi; Bronchoconstriction; Dose-Response Relationship, Drug; Enzyme Activation; Enzyme Inhibitors; Flavonoids; Guinea Pigs; Histamine Release; Hypersensitivity; Immunoglobulin G; In Vitro Techniques; Leukotriene D4; Lung; Male; Mast Cells; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase Kinases; Ovalbumin; Protein Kinase Inhibitors; Protein Kinases

The purpose of this study was to explore whether repeated exposure to aerosolized ovalbumin (OVA) in the context of local expression of GM-CSF can initiate a Th2-driven, eosinophilic inflammation in the airways. On day -1, Balb/c mice were infected intranasally with an adenovirus construct expressing GM-CSF (Ad/GM-CSF). From day 0 to day 9 mice were exposed daily to an OVA aerosol. Mice exposed to OVA alone did not show any evidence of airway inflammation. Mice receiving both Ad/GM-CSF and aerosolized OVA exhibited marked airway inflammation characterized by eosinophilia and goblet cell hyperplasia. Migration of eosinophils into the airway was preceded by a rise in IL-5 and IL-4. Both IL-5 and class II MHC were critically required to generate airway eosinophilia. After resolution, airway eosinophilia was reconstituted after a single OVA exposure. Flow cytometric analysis of dispersed lung cells revealed an increase in macrophages and dendritic cells expressing B7.1 and B7.2, and expansion of activated (CD69-expressing) CD4 and CD8 T cells in mice exposed to OVA and Ad/GM-CSF. Our data indicate that expression of GM-CSF in the airway compartment increases local antigen presentation capacity, and concomitantly facilitates the development of an antigen-specific, eosinophilic inflammatory response to an otherwise innocuous antigen.

Topics: Adenoviruses, Human; Aerosols; Allergens; Animals; Antigens; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Dendritic Cells; Eosinophils; Female; Gene Expression; Genetic Vectors; Granulocyte-Macrophage Colony-Stimulating Factor; Histocompatibility Antigens Class II; Humans; Hypersensitivity; Immunity, Mucosal; Interleukin-4; Interleukin-5; Lung; Macrophages; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Time Factors; Transgenes

The inflammatory response as seen in human allergic asthma is thought to be regulated by Th2 cells. It has been shown that interferon-gamma (IFN-gamma) can downregulate the proliferation of Th2 cells and therefore might be of therapeutic use. In the present study we have investigated the in vivo role of endogenous and exogenous IFN-gamma in a murine model with features reminiscent of human allergic asthma. IFN-gamma gene knockout (GKO) and wild-type mice were sensitized with ovalbumin and exposed to repeated ovalbumin aerosol challenges. In addition, wild-type mice were treated with intraperitoneal or nebulized recombinant murine IFN-gamma during the challenge period. Sensitized wild-type mice exhibited upregulated ovalbumin-specific IgE in serum, and airway hyperresponsiveness and infiltration of eosinophils and mononuclear cells in the bronchoalveolar lavage fluid (BALF) after ovalbumin challenge. In contrast, in GKO mice only reduced eosinophilic infiltration in the BALF was observed after ovalbumin challenge. In wild-type mice, parenteral IFN-gamma treatment downregulated ovalbumin-specific IgE levels in serum, and airway hyperresponsiveness and cellular infiltration in the BALF, whereas aerosolized IFN-gamma treatment only suppressed airway hyperresponsiveness. In vitro experiments showed that these effects of IFN-gamma appear not to be mediated via a direct effect on the cytokine production of antigen-specific Th2 cells. These data indicate that airway hyperresponsiveness can be downregulated by IFN-gamma locally in the airways, whereas for downregulation of IgE and cellular infiltration systemic IFN-gamma is needed. The present study shows that exogenous IFN-gamma can downregulate the allergic response via an antigen-specific T-cell independent mechanism, but at the same time endogenous IFN-gamma plays a role in an optimal response.

Topics: Administration, Inhalation; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Hypersensitivity; Immunoglobulin E; Injections, Intraperitoneal; Interferon-gamma; Leukocytes; Lymph Nodes; Macrophages, Alveolar; Methacholine Chloride; Mice; Mice, Knockout; Ovalbumin; Spleen; T-Lymphocytes

The role of nerve growth factor (NGF), a potent mediator acting in the development and differentiation of both neuronal and immune cells, was examined in a mouse model of allergic asthma. NGF-positive cells were detected in the inflammatory infiltrate of the lung and enhanced levels of NGF were detected in serum and broncho-alveolar lavage fluids. Mononuclear cells in inflamed airway mucosa as well as broncho-alveolar macrophages were identified as one source of NGF production. Splenic mononuclear cells from allergen-sensitized mice produced NGF in response to allergen. They responded to exogenously added NGF with a dose-dependent increase in IL-4 and IL-5 production and augmented IgE and IgG1 synthesis. In contrast, IFN-gamma and IgG2alpha levels remained unaffected. The effects were NGF specific, since they could be blocked by an anti-NGF-antibody. Nasal application of anti-NGF to allergen-sensitized mice significantly reduced IL-4 and prevented development of airway hyperreactivity. These results show that allergic airway inflammation is accompanied by enhanced local NGF production that acts as an amplifier for Th2 effector functions and plays an important role in the development of airway hyperreactivity. Therefore it is suggested that NGF may serve as a link between the immune and nerve system.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Cells, Cultured; Disease Models, Animal; Hypersensitivity; Inflammation; Leukocytes, Mononuclear; Mice; Mice, Inbred BALB C; Nerve Growth Factors; Ovalbumin; Th2 Cells

Strength of T cell receptor (TCR) signaling, coreceptors, costimulation, antigen-presenting cell type, and cytokines all play crucial roles in determining the efficiency with which type 2 T lymphocytes (Th2, Tc2) develop from uncommitted precursors. To investigate in vivo regulatory mechanisms that control the population of type 2 T cells and disease susceptibility, we have created lines of transgenic mice in which expression of a chimeric cytokine receptor (the mouse interleukin 2 receptor beta chain [IL-2Rbeta] extracellular domain fused to the cytoplasmic tail of IL-4Ralpha) is targeted to the T lymphoid lineage using the proximal lck promoter. This chimera transduced IL-4-specific signals in response to IL-2 binding and dramatically enhanced type 2 responses (IL-4, IL-5, and immunoglobulin E production) upon in vitro TCR stimulation or in vivo antigen challenge. Thus, type 2 effector function was augmented by IL-4 signals transduced through a chimeric receptor expressed in a T cell-specific manner. This influence was sufficient for establishment of antigen-induced allergic airway hyperresponsiveness on a disease-resistant background (C57BL/6).

Topics: Animals; Asthma; Bronchial Hyperreactivity; Flow Cytometry; Humans; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Methacholine Chloride; Mice; Mice, Transgenic; Ovalbumin; Receptors, Interleukin-2; Receptors, Interleukin-4; Recombinant Fusion Proteins; Signal Transduction; STAT6 Transcription Factor; T-Lymphocytes; Th2 Cells; Trans-Activators

Human foods are usually prepared by cooking. Boiling of chicken egg-white (EW) led to decreased allergenicity, and abrogated intestinal uptake of immunoreactive ovalbumin (OVA) when fed to mice. Therefore, the effects of oral administration of boiled EW were examined further in BALB/c mice. Specific IgE, IgG1 and IgG antibody responses were suppressed by raw EW, but not by EW boiled for 5 or 60 min, fed prior to sensitization with 10 microg OVA or 1 microg DNP-OVA in alum. Similar results were obtained when mice were sensitized with 10 microg conalbumin, ovomucoid or lysozyme in alum. BALB/c spleen cell proliferation and secretion of Th2 cytokines IL-4 and IL-5 during in vitro stimulation with OVA were also suppressed by feeding raw EW, but not by boiled EW. Although heat denaturation of proteins can minimize allergenicity, the present results suggest that over-cooking of proteins may affect their intestinal antigen processing and thus prevent the induction of oral tolerance.

Topics: Animals; Cell Division; Conalbumin; Dinitrobenzenes; Egg Proteins; Female; Heating; Hypersensitivity; Immune Tolerance; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Muramidase; Ovalbumin; Ovomucin; Protein Denaturation; Rats; Rats, Sprague-Dawley; Spleen; Th2 Cells

The effect of systemic and local allergic responses on the mucociliary system of the Eustachian tube was investigated. Egg albumin was administered to guinea pigs via the jugular vein to evoke systemic anaphylaxis in animals previously sensitized with egg albumin. Ciliary activity in the Eustachian tube of sensitized animals was significantly higher than that of control animals. However, the mucociliary clearance time of the Eustachian tube in sensitized animals was significantly longer than in control animals. Local allergic response induced by intratympanic instillation of (BPO)61-BGG induced cilioexcitation and prolonged mucociliary clearance in the guinea pigs sensitized with BPO-BGG. A partial loss of the inner layer of the mucus blanket of the Eustachian tube was observed under electron microscopy. In conclusion, the systemic as well as the local allergic response accelerates ciliary activity but may affect the mucus blanket and induce mucociliary dysfunction in the Eustachian tube, resulting in a predisposition to middle ear diseases.

Topics: Allergens; Anaphylaxis; Animals; Benzeneacetamides; Cilia; Eustachian Tube; Female; gamma-Globulins; Guinea Pigs; Hypersensitivity; Male; Mucociliary Clearance; Ovalbumin; Penicillin G

Topics: Animals; Chloride Channels; Chlorides; Electric Conductivity; Epithelium; Guinea Pigs; Hypersensitivity; Intestinal Mucosa; Mucous Membrane; Ovalbumin; Trachea

Optimal treatment of allergic diseases requires that the cytokine profile of allergen-specific T cells be redirected, with the conversion of Th2 profiles into Th1 cytokine profiles. This conversion, however, is difficult, since Th2 effector cells have relatively fixed cytokine profiles. To more effectively redirect the cytokine profiles of T cells, we constructed a cytokine fusion protein that contained the Ag OVA, fused to IL-12. Immunization with the OVA-IL-12 fusion protein induced anti-OVA IgG2a Ab and large quantities of OVA-specific IFN-gamma production. The Ag specificity of this response was dependent upon covalent linkage of Ag and IL-12, since immunization of mice with OVA alone induced little or no IFN-gamma, while immunization with OVA and free rIL-12 enhanced T cell production of IFN-gamma, but the IFN-gamma production was not OVA specific. To examine the effects of OVA-IL-12 in reversing ongoing Th2-dominated immune responses, BALB/c mice previously primed with OVA in alum to induce a Th2-dominated response, were vaccinated with the OVA-IL-12 protein. In such mice, OVA-IL-12 was much more effective than OVA plus free rIL-12 in significantly increasing Ag-specific IFN-gamma production and significantly decreasing Ag-specific IL-4 production. Moreover, OVA-IL-12 increased serum anti-OVA IgG2a and decreased anti-OVA IgE. These studies indicate that OVA-IL-12 can convert immune responses characterized by high IL-4 and high IgE synthesis into Th1-dominated responses in an Ag-specific manner.

Topics: Animals; Antigens; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-12; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Fusion Proteins; Th1 Cells; Th2 Cells

Monocyte chemotactic protein-3 (MCP-3)/fibroblast-induced cytokine (FIC), a CC chemokine, is chemotactic for cells that typically infiltrate the late-phase allergic reaction. We developed a mouse model of airway inflammation to study the role of MCP-3/FIC. The immunization of mice with OVA resulted in Ag-specific IgE Ab production and the expression of mRNA for IL-4 in the lung tissue. Two weeks after immunization mice were challenged with the allergen by inhalation. Lungs were lavaged, and the tissue was examined at 2 or 24 h. Allergen challenge resulted in the increased recovery of leukocytes in the lavage fluid, but saline challenge did not. There was a significant increase in eosinophils (29 +/- 8% vs 1.2 +/- 0.2%) and lymphocytes (25 +/- 4% vs 5 +/- 2%) in the bronchoaveolar lavage fluid. Histologic examination of the lung demonstrated intense airway inflammation following OVA challenge. The expression of MCP-3/FIC and other CC chemokines (MCP-1, macrophage inflammatory protein-1alpha, and RANTES) was investigated by reverse transcription-PCR followed by densitometric analyses. The allergen challenge up-regulated the expression of mRNA for MCP-1, MCP-3/FIC, and macrophage inflammatory protein-1alpha at 2 and/or 24 h. Immunocytochemical staining for MCP-3/FIC showed that the allergen challenge induced the expression of MCP-3/FIC predominantly in the airway epithelium. Pretreatment of mice with an anti-MCP-3/FIC Ab significantly inhibited the OVA-induced airway inflammation and the bronchoalveolar lavage eosinophilia (8 +/- 2% vs 46 +/- 11% after control Ab, p < 0.03). We conclude that MCP-3/FIC plays a significant role in the allergen-induced eosinophilic inflammation of the airways.

Topics: Animals; Asthma; Chemokine CCL2; Chemokine CCL4; Chemokine CCL5; Chemokine CCL7; Chemokines; Cytokines; Hypersensitivity; Interleukin-4; Lung; Macrophage Inflammatory Proteins; Mice; Mice, Inbred BALB C; Monocyte Chemoattractant Proteins; Ovalbumin; RNA, Messenger

Viral respiratory infections can predispose to the development of asthma by mechanisms that are presently undetermined. Using a murine model of respiratory syncytial virus (RSV) infection, acute infection is associated with airway hyperresponsiveness as well as enhanced responses to subsequent sensitization to allergen. We demonstrate that acute viral infection results in increased airway responsiveness to inhaled methacholine and pulmonary neutrophilic and eosinophilic inflammation. This response is associated with predominant production of Th-1-type cytokines in peribronchial lymph node cells in vitro. Mice sensitized to ovalbumin via the airways after RSV infection developed increased airway responsiveness to methacholine and pulmonary eosinophilic and neutrophilic inflammation, associated with the predominant production of Th-2-type cytokines. Treatment of the mice with anti-IL-5 antibody abolished airway hyperresponsiveness and eosinophilic but not neutrophilic inflammation in both acutely infected mice and mice sensitized after infection. We conclude that RSV infection results in airway hyperresponsiveness in the acute phase and leads to changes in immune function that can enhance the effects of airway sensitization to antigen after infection. In both situations, airway hyperresponsiveness is closely associated with pulmonary eosinophilic inflammation. This model provides a means for further analyzing the influence of viral respiratory infections on airway sensitization and the development of altered airway responsiveness.

Topics: Allergens; Animals; Bronchoconstrictor Agents; Cytokines; Eosinophils; Female; Humans; Hypersensitivity; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-5; Methacholine Chloride; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human; Th2 Cells; Time Factors; Tumor Cells, Cultured

Neuronal M2 muscarinic receptors inhibit acetylcholine release from pulmonary parasympathetic nerves but are dysfunctional in antigen-challenged animals and asthmatics. Deletion of pulmonary eosinophils protects M2 receptor function in antigen-challenged guinea pigs. Therefore, the association of eosinophils with airway nerves was investigated. Nerve-associated eosinophils were significantly increased in challenged animals compared with controls (0.75 +/- 0.05 vs. 0.28 +/- 0.05 eosinophils/nerve). In antigen-challenged animals, eosinophil density was greatest around airway nerves, suggesting recruitment to the nerves. M2 receptor function was inversely correlated with the number of eosinophils per nerve, thus eosinophils are associated with airway nerves in antigen-challenged guinea pigs, where they impair M2 receptor function. In airways from three patients with fatal asthma, 196 of 637 eosinophils (30%) were associated with nerves, and release of eosinophil major basic protein was evident; conversely, in three control patients 1 of 11 (9%) eosinophils were in contact with nerves. Thus eosinophils and their granule proteins are also seen in association with airway nerves in patients with asthma.

Topics: Acetylcholinesterase; Adult; Animals; Antigens; Asthma; Eosinophils; Female; Guinea Pigs; Humans; Hypersensitivity; Lung; Male; Middle Aged; Neurons; Ovalbumin; Parasympathetic Nervous System; Pilocarpine; Receptor, Muscarinic M2; Receptors, Muscarinic; Vagus Nerve

Respiratory syncytial virus (RSV) causes acute bronchiolitis in children and has been implicated in the pathogenesis of recurrent wheezing and asthma. However, few children exposed to RSV experience acute bronchiolitis or its sequelae, suggesting a subgroup of susceptible children. An allergic diathesis may predispose children to subsequent airway disease.. This study was carried out to determine whether a preexisting allergic state, induced by repeated inhalational exposures to ovalbumin, potentiates nonspecific airway responsiveness to acetylcholine and increases airway inflammation during acute RSV bronchiolitis in guinea pigs.. Forty guinea pigs were randomized into four groups: nonsensitized, noninfected (ovalbumin-, RSV-); sensitized, noninfected (ovalbumin+, RSV-); nonsensitized, infected (ovalbumin-, RSV+); sensitized, infected (ovalbumin+, RSV+). Depending on grouping, animals were exposed to either repeated aerosols of ovalbumin or saline solution and were subsequently inoculated with either human RSV or uninfected culture medium. Six days after inoculation, animals underwent acetylcholine challenge, and lung specimens were prepared for histologic scoring of airway inflammation.. Maximal increases in pulmonary resistance (centimeters of water per milliliter per second) to acetylcholine were greater for RSV alone (12.4 +/- 3.9) and ovalbumin alone (13.7 +/- 3.9) compared with controls (4.3 +/- 1.1), but significantly greater increases occurred in ovalbumin+, RSV+ animals (34.0 +/- 11.0). These ovalbumin+, RSV+ animals demonstrated the combined histologic changes noted with RSV alone and ovalbumin alone including airway epithelial necrosis, mononuclear and granulocyte infiltrates, airway wall edema, hyperplasia of bronchus-associated lymphoid tissue, and goblet cell metaplasia.. Prior allergic sensitization potentiates the physiologic and structural changes induced by acute RSV bronchiolitis. These results suggest that an allergic diathesis may increase the severity of RSV infections in children.

Topics: Acetylcholine; Administration, Inhalation; Animals; Bronchial Hyperreactivity; Bronchiolitis, Viral; Guinea Pigs; Humans; Hypersensitivity; Immunization; Ovalbumin; Respiratory Syncytial Virus Infections

In studies of disease processes, increasing knowledge leads to an increased awareness of the complexity of the underlying mechanisms. The intense research activity in the chemokine field has made this acutely manifest. Numerous chemokines have been discovered through the use of (1) bioassay of in vitro cell culture supernatants and in vivo exudates from animal models of inflammation and (2) molecular biology techniques. Any one chemokine can often be produced by a number of different cell types and exert its effects on different target cells. This has been interpreted by some as implying a high degree of redundancy. Although this is understandable, in disease processes parallel and sequential mechanisms are possible, and potentially important therapeutic targets have emerged. There is compelling evidence from animal and clinical studies that eosinophils are important effector cells in asthma, but this relationship is as yet unproven in the human disease. Two possible targets to prevent eosinophil recruitment to the lung are IL-5 and its receptor, which are important in several aspects of eosinophil biology, and eotaxin and its receptor, CCR3. The eotaxin receptor is particularly attractive as a target as it is expressed in high numbers on eosinophils, but not other leukocytes, and appears to be the major detector of the eosinophil for eotaxin and other chemokines such as MCP-4. Eotaxin and CCR3 knockout mice are being developed, and animal models will continue to be invaluable when antagonists are available. In the shape of receptor antagonists, the chemokine field may yet provide the final proof of concept for the long-established eosinophil theory of asthma in humans.

Topics: Amino Acid Sequence; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokine CCL2; Chemokine CCL5; Chemokines; Chemokines, CC; Chromatography, High Pressure Liquid; Cytokines; Disease Models, Animal; Endothelium, Vascular; Humans; Hypersensitivity; Interleukin-8; Leukocytes; Molecular Sequence Data; Ovalbumin

1 The aim of the present research was to study the cholinergic and inhibitory non-adrenergic-non-cholinergic (NANC) responses obtained with electrical field stimulation (EFS) of tracheal tissues from sham- and noise-exposed guinea-pigs. A comparison was also made between normal and ovalbumin (OA)-sensitized animals. 2 In proximal tracheae pretreated with indomethacin (3 microM), propranolol (1 microM), alpha-chymotrypsin (2 U ml-1) and L-NAME (0.1 mM), frequency-dependent responses to EFS (0.1 ms width; 20 V, 0.1-100 Hz, 15 s train duration) were obtained, both contractile and relaxing in nature. The contractile responses were abolished by atropine (1 microM), and did not vary significantly between sham- and noise-exposed guinea-pigs, or between normal and sensitized animals. The NANC relaxing responses, present in spite of the pre-treatment of the tissues with L-NAME and alpha-chymotrypsin, and almost completely abolished by tetrodotoxin (TTX) treatment (10 microM), appeared to be enhanced in noise-exposed guinea-pigs, with respect to sham-exposed animals, but only when the animals were not OA-sensitized. 3 In distal tracheae contracted with histamine (10 microM), the study of the whole inhibitory NANC response (pre-treatment with propranolol, but not with alpha-chymotrypsin and L-NAME), which was mainly TTX-sensitive, revealed a statistically non-significant difference between sham- and noise-exposed guinea-pigs, both normal and OA-sensitized. When distal tracheae were preincubated with alpha-chymotrypsin (2 U ml-1) and L-NAME (0.1 mM), in addition to propranolol, a significant residual inhibitory NANC response to EFS was observed. Surprisingly, in this case, similarly to the evidence obtained in proximal tracheae, a significantly enhanced response was revealed in noise-exposed guinea-pigs with respect to sham-exposed animals. 4 The noise-induced enhancement of the relaxant response disappeared when the tissues were pretreated with the A2 purinergic antagonist 3,7-dimethyl-1-propargylxanthine (DMPX, 1 microM), while it persisted in the presence of the A1 antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, 10 nM). 5 The above data indicate that, while not modifying the cholinergic and the whole inhibitory NANC response to EFS, noise stress selectively influences an inhibitory component of the NANC system in guinea-pig trachea with a mechanism probably involving an enhanced neurally mediated release of adenosine, which relaxes the smooth muscle via A2 r

Topics: Acetylcholine; Animals; Autonomic Nervous System; Electric Stimulation; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Male; Muscle Contraction; Muscle Relaxation; Muscle, Smooth; Neural Inhibition; Noise; Ovalbumin; Receptors, Purinergic P1; Stress, Physiological; Trachea

We summarize here the main characteristics of a novel model of pulmonary hypersensitivity. Mice were immunized with a subcutaneous implant of a fragment of heat solidified chicken egg white and 14 days later challenged with ovalbumin given either by aerosol or by intratracheal instillation. This procedure induces a persistent eosinophilic lung inflammation, a marked bone marrow eosinophilia, and Th2-type isotypic profile with histopathological findings that resemble human asthma. Further, this model is simple to perform, reproducible in different strains of mice, does not require adjuvants nor multiple boosters. Based on these characteristics we propose it as a suitable murine model of allergic eosinophilic lung inflammation.

Topics: Animals; Asthma; Disease Models, Animal; Eosinophils; Hypersensitivity; Inflammation; Lung; Mice; Ovalbumin; Pulmonary Eosinophilia

A method to introduce 111In-labelled neutrophils or eosinophils into the circulation of anaesthetized, ovalbumin-sensitized guinea pigs and monitor their pulmonary accumulation using gamma scintigraphy has been developed. The method is based on the ability to use 99mTc macroaggregated albumin (MAA) to create a pulmonary perfusion image as a template for the lungs of individual guinea pigs which can be superimposed on to the image produced by the 111In-labelled leukocytes injected into the same animal. Intravenous injection of the labelled leukocytes was shown to produce a high density radioactivity in the heart and lungs with little circulation to the rest of the body. This suggested an immediate 'trapping' of leukocytes in the pulmonary vascular bed. A more effective distribution of 111In-labelled cells was achieved by injecting them into the right carotid artery. This ensured more efficient circulation of the cells and resulted in their appearance in the lungs, liver and spleen. However, it was found that the contralateral carotid artery had to be tied off to prevent the cells only circulating to the superior left quadrant of the animal. Bronchoalveolar lavage of the lungs following i.v. injection of 111In-labelled neutrophils showed no significant difference in radioactivity of lavage fluid between guinea pigs challenged 24 h beforehand with inhaled saline or ovalbumin. In contrast, when labelled eosinophils were injected, there was a significantly greater mean radioactivity level in lavage fluid after ovalbumin challenge than after saline. In conclusion, gamma scintigraphy provides an accurate measure of the amount of activity from 111In-labelled leukocytes in the lung region which is specific for each animal

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Movement; Eosinophils; Guinea Pigs; Hypersensitivity; Indium Radioisotopes; Lung; Neutrophils; Ovalbumin; Radionuclide Imaging

Transcription factors of the NFAT family are thought to play a major role in regulating the expression of cytokine genes and other inducible genes during the immune response. The role of NFAT1 was investigated by targeted disruption of the NFAT1 gene. Unexpectedly, cells from NFAT1 -/- mice showed increased primary responses to Leishmania major and mounted increased secondary responses to ovalbumin in vitro. In an in vivo model of allergic inflammation, the accumulation of eosinophils and levels of serum immunoglobulin E were increased in NFAT1 -/- mice. These results suggest that NFAT1 exerts a negative regulatory influence on the immune response.

Topics: Amino Acid Sequence; Animals; Antigens, Protozoan; Cell Line; Cytokines; DNA-Binding Proteins; Eosinophils; Gene Targeting; Hypersensitivity; Immunity; Immunoglobulin E; Immunologic Memory; Leishmania major; Lymphocyte Activation; Mice; Molecular Sequence Data; NFATC Transcription Factors; Nuclear Proteins; Ovalbumin; T-Lymphocytes; Transcription Factors

We experimentally demonstrated the anti-asthmatic effects of M-711, the dry extract of a Chinese herb medicine called Mokuboi-to, using the rat model of allergic asthma. Allergic asthma was induced by the antigen-antibody reaction in the rats and they showed anaphylactic symptoms accompanied with hypotension and depression of respiratory function. More than 20 mg/kg body weight of M-711 was effective in relieving the asthmatic symptoms like methylephedrine. It could lessened the suppress of respiration and provided a good improvement. It also reduced a drop of the blood pressure and improved it quickly. Its ingredients alone were much less effective than M-711 as the total. Those data suggested that M-711 was effective for alleviating allergic asthma in the present study and that its action was provided by interaction of all ingredient raw herbs of M-711.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Blood Pressure; Bronchitis; Calcium Sulfate; Cinnamomum zeylanicum; Dog Diseases; Dogs; Drugs, Chinese Herbal; Hypersensitivity; Medicine, Chinese Traditional; Ovalbumin; Panax; Phytotherapy; Plant Extracts; Plants, Medicinal; Rats; Respiratory Function Tests; Time Factors

We determined the expression of Th-2 type cytokines, interleukin-4 (IL-4) and IL-5, and of the Th-1 type cytokine, interferon-gamma (IFN-gamma), in the Brown-Norway rat. Rats were intraperitoneally sensitized with ovalbumin and 21 days later were either exposed to ovalbumin or saline aerosol. The value -log PC300 (PC300 = concentration of acetylcholine needed to increase baseline lung resistance by 300%) was 2.49 +/- 0.15 in sensitized, exposed rats, was higher than in sensitized, saline-exposed or naive rats (1.54 +/- 0.27 and 1.63 +/- 0.06 respectively, P < 0.05). There was a significant increase in eosinophils in bronchoalveolar lavage fluid and in airway submucosal airway tissues in the sensitized exposed group. Reverse-transcriptase polymerase chain reaction was performed on total lung RNA using primers for IL-4, IL-5, IFN-gamma and beta-actin. IL-4 and IL-5 mRNA levels in control and sensitized saline-exposed rats were not detectable, but increased levels were found in sensitized and ovalbumin-exposed rats with levels of 0.25 +/- 0.01 and 0.98 +/- 0.02% of beta-actin mRNA as assessed by densitometric measurements. Expression of IFN-gamma mRNA was significantly reduced in sensitized and ovalbumin-exposed rats. As in asthmatic airways, there is an increased expression of Th-2 cytokines, IL-4 and IL-5, together with a reduction in the Th-1 cytokine, IFN-gamma, thus supporting a role for Th-2 cytokines in allergic eosinophilic inflammation.

Topics: Animals; Disease Models, Animal; Hypersensitivity; Immunohistochemistry; Interferon-gamma; Interleukin-4; Interleukin-5; Lung; Male; Ovalbumin; Polymerase Chain Reaction; Rats; Rats, Inbred BN; RNA, Messenger; T-Lymphocytes, Helper-Inducer

Cells producing IgA specific to ovalbumin (OVA) were detected with an assay of plaque-forming cells (PFC). Non-T cells were separated on a polystyrene resin column and were further depleted of B cells that bound sheep erythrocytes (SRBC) by SRBC-rosette sedimentation. The cells were recombined with T cells separated on a polystyrene resin column, stimulated with OVA antigen, and then cultured for 5 d. The number of OVA-specific IgA-PFC from the lymphocytes of infants allergic to hen's eggs (7 +/- 5 per 7 x 10(4) non-T cells, n = 9) was significantly less than that of PFC from lymphocytes of age-matched controls (110 +/- 18 per 7 x 10(4) non-T cells, n = 7) and from those of children with atopic dermatitis who were not allergic to hen's eggs (90 +/- 30 per 7 x 10(4) non-T cells, n = 4). Patients' B cells added to the culture supernatant from OVA-stimulated normal T cells (82 +/- 18 per 7 x 10(4) non-T cells, n = 4) were able to produce the specific IgA to levels comparable to those of normal B cells (92 +/- 9 per 7 x 10(4) non-T cells, n = 6), but the patients' T cells did not cause normal B cells to produce the antibody (8 +/- 2 per 7 x 10(4) non-T cells, n = 4). This indicates that the patients' T cells were less able than were normal T cells to promote the production of OVA-specific IgA-PFC. Until the age of 6 y, the ability of the patients' lymphocytes to produce specific IgA was abnormally low; from that age on, it was normal. At the stage of allergen entry, this transiently low production of OVA-specific IgA may contribute to the onset of allergy to hen's eggs.

Topics: Cells, Cultured; Child; Child, Preschool; Female; Humans; Hypersensitivity; Immunoglobulin A; Infant; Male; Ovalbumin; Ovum; T-Lymphocytes

In atopic patients, allergen-sensitized T lymphocytes specifically proliferate in the presence of T cell-growth factor, interleukin 2 (IL-2). Lecithin-bound iodine (LBI), which has been used as a therapeutic modality for patients with bronchial asthma (BA), effectively inhibited Dermatophagoides farinae (Df) mite antigen-induced IL-2 responsiveness in concentrations comparable to LBI concentrations in blood. IL-2-responding T cells were more sensitive to LBI than antigen-presenting cells, whereas LBI suppressed the release of interleukin 1 (IL-1) elicited by Df antigen. In addition, ovalbumin (OVA)-induced IL-2 responsiveness in egg sensitive patients and purified protein-derivative (PPD)-induced IL-2 responsiveness were similarly inhibited by LBI. The IL-2 responsiveness induced by concanavalin A (Con A), however, was not changed. On the basis of these results, LBI may act as a slight immunosuppressant to inhibit the induction of allergen-specific lymphocytes and to improve the clinical status in allergic diseases.

Topics: Adult; Allergens; alpha-Linolenic Acid; Antigens, Dermatophagoides; Asthma; Child, Preschool; Concanavalin A; Dermatitis, Atopic; Epitopes; Glycoproteins; Humans; Hypersensitivity; Infant; Interleukin-1; Interleukin-2; Iodine; Lymphocyte Activation; Ovalbumin; Phosphatidylcholines; T-Lymphocytes; Tuberculin

Current evidence suggests that the events subsequent to antigen challenge in allergic asthmatics involve eosinophil activation and the synthesis of proinflammatory cytokines, in particular interleukin 5 (IL-5). However, little is known about how local inflammatory cell infiltration and activation are related to the changes in lung function following allergen exposure. We have developed a novel technique to investigate the local inflammatory events during late-onset allergic bronchoconstriction in lung explants from sensitized Brown-Norway (BN) rats. In this study we tested the hypothesis that the in vitro late airway response involves IL-5 gene activation and recruitment and activation of eosinophils. Explants were prepared from excised lungs of BN rats (n = 9) sensitized 2 wk previously to ovalbumin (OVA). Lungs were inflated with liquid agarose solution (2% wt/vol, 48 ml/kg) following perfusion with cold Ca2+/Mg(2+)-free Hanks' solution, and refrigerated briefly to gel the agarose, and 0.5- to 1.0-mm slices were prepared and cultured overnight at 37 degrees C. Airways were identified and challenged by direct application of OVA (20 micrograms). Cryostat sections of explants were immunostained for major basic protein (MBP) and IL-5 mRNA was detected by a 35S-uridine triphosphate-labeled probe and in situ hybridization. Explants harvested immediately prior to challenge showed little evidence of MBP and IL-5 mRNA expression. Explants harvested at 6 h which exhibited evidence of bronchoconstriction showed strong cell-associated immunostaining for MBP and high expression of IL-5 mRNA in the bronchial mucosa. colocalization studies performed in lung explants demonstrating late-onset airway responses suggested that the majority of IL-5 mRNA expression was not found in MBP-positive cells. When compared with explants from sham-sensitized rats (n = 4), there was a significant increase in MBP-positive and IL-5 mRNA-positive cells per millimeter of basement membrane of the airway. The presence of MBP immunoreactivity and IL-5 gene expression was not observed in explants taken from sensitized BN rats which did not undergo late-onset airway responses, indicating an association between inflammatory cell activation and airway constriction. The increase in MBP-positive cells several hours after OVA suggests activation, local recruitment, and/or differentiation of eosinophils. This study provides direct evidence for a temporal association between IL-5 expression, eosinoph

Topics: Allergens; Animals; Basement Membrane; Blood Proteins; Bronchoconstriction; Eosinophil Granule Proteins; Eosinophils; Gene Expression Regulation; Hypersensitivity; Interleukin-5; Lung; Ovalbumin; Rats; Rats, Inbred BN; Ribonucleases; RNA, Messenger; Transcriptional Activation

MDL 105,212 has been identified as a potent, nonpeptide NK-1 and NK-2 receptor antagonist that inhibits effects of substance P and neurokinin A in vitro and in vivo (Kudlacz et al., 1996). In the present study, the compound inhibited capsaicin-induced respiratory effects after p.o. administration (5-50 mg/kg) to conscious guinea pigs; nearly complete inhibition of dyspnea and cough was observed 1 hr after 50 mg/kg p.o., and efficacy persisted for approximately 11 hr. MDL 105,212 reduced pulmonary insufflation pressure and microvascular leakage in ovalbumin-sensitized animals in response to antigen-challenge relative to vehicle-treated animals. Attenuation of early-phase allergic responses may result from MDL 105,212 inhibition of antigen-induced histamine release from sensitized guinea pig lung observed in vitro. Airway hyperresponsiveness to methacholine occurred 24 hr after antigen-challenge in ovalbuminsensitized guinea pigs; this effect was inhibited by pretreatment with MDL 105,212 (50 mg/kg p.o.) 1 hr before ovalbumin exposure without affecting increased bronchoalveolar lavage eosinophil numbers. These data suggest that sensory neuropeptides play a role in some aspects of allergic airway responses and that tachykinin receptor antagonists may be useful in treatment of atopic respiratory diseases.

Topics: Animals; Bronchoconstriction; Capsaicin; Dose-Response Relationship, Drug; Guinea Pigs; Histamine Release; Hypersensitivity; Male; Neurokinin-1 Receptor Antagonists; Ovalbumin; Piperidines; Pyrrolidines; Receptors, Neurokinin-2; Respiration

Arachidonic acid (AA), the precursor of eicosanoids, is released from the sn-2 position of phospholipids by both secretory (sPLA2) and cytosolic phospholipase A2 (cPLA2). Eicosanoids have been shown to contribute to bronchospasm in asthma. We measured the enzymatic activity of sPLA2 and cPLA2 in the bronchoalveolar lavage fluid and cells, respectively, in male Hartley guinea pigs sensitized with ovalbumin. sPLA2 activity was also measured from alveolar macrophages (AM) in culture from unsensitized and sensitized animals. There was an increase in sPLA2 activity and AA content in the lavage fluid following sensitization (18.73 +/- 1.33 to 25.74 +/- 3.22% hydrolysis and 17.97 +/- 12.39 to 44.76 +/- 13.37 pmol AA/mL BAL, mean +/- SD), which remained elevated but without further increase 4 or 24 h after antigen challenge. AM from unsensitized and sensitized-unchallenged animals did not secrete sPLA2 activity in culture for 3 h and therefore do not appear to be the cell source of the sPLA2 activity present in the alveolar lavage fluid following OA sensitization. In contrast to the increase in sPLA2 in lung lavage fluid, Western blotting for cPLA2 from lung lavage cells showed no increase 4 or 24 h after antigen challenge compared with sensitization alone. cPLA2 enzymatic activity of the cytosol fraction of lung lavage cells showed no changes with antigen sensitization or challenge. In summary, intraperitoneal sensitization with ovalbumin in male Hartley guinea pigs caused an increase in both sPLA2 and AA in bronchoalveolar lavage fluid without a need for antigen challenge. The increased sPLA2 enzymatic activity following sensitization may be responsible for the elevation of AA in the bronchoalveolar lavage fluid observed after antigen sensitization.

Topics: Animals; Arachidonic Acid; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Guinea Pigs; Hypersensitivity; Macrophages, Alveolar; Male; Ovalbumin; Phospholipases A; Phospholipases A2; Time Factors

Upper respiratory tract viral infections have been reported in clinical studies to serve as risk factors for allergic sensitization. In order to study the relationship linking influenza virus illnesses to development of allergy, murine models of allergen sensitization were previously employed. These models showed that lethal influenza viruses were able to trigger allergen-specific immunoglobulin E (IgE) production and to inhibit tolerance to repeated exposure to aerosolized allergen in the mouse. The disadvantage of these murine models consists in the utilization of virulant and lethal strains of influenza virus. A nonlethal rat-adapted influenza virus (RAIV) host resistance model has been developed in our laboratory. It was used to evaluate the effect of influenza virus infection on IgE responses to inhaled ovalbumin (OA) in the rat. The high IgE-responder Brown-Norway (BN) rat was chosen for further study after comparing the IgE response to OA in Fischer 344 (F344) and BN rats. On d 1, BN rats were sensitized by administration of 1 mg OA subcutaneously alone or together with aluminum hydroxide (200 mg) and Bordetella pertussis (15 x 10(9) killed bacilli per rat in 1 ml), or only received saline. Rats were either infected with RAIV or sham-infected on d 0 (24 h prior to sensitization) or on d 15, 17, or 57. Rats were exposed for 3 min to aerosolized OA (OA 3% in phosphate-buffered saline) every week, starting on d 18. Serum OA-specific IgE was evaluated by reverse enzyme-linked immunosorbent assay (ELISA) 3 d after each OA challenge. BN rats elicited a detectable OA-specific IgE response that decreased after repeated aerosol exposures. Influenza virus infection transiently increased the OA-specific IgE response when rats were immunized with OA alone and were infected 1 d prior to the first challenge and also when rats received only saline on d 1, were exposed each week to aerosolized OA, and were infected prior to the seventh challenge. These results, with data previously reported in mice, emphasize the importance of upper respiratory tract viral infection in increasing IgE responses to allergens and may be of importance in human disease.

Topics: Administration, Inhalation; Aerosols; Animals; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Injections, Subcutaneous; Orthomyxoviridae; Orthomyxoviridae Infections; Ovalbumin; Rats; Rats, Inbred BN; Rats, Inbred F344; Risk Factors; Viral Plaque Assay; Virus Replication

Activation of naive T cells requires at least two signals. In addition to the well characterized interaction of the T cell antigen receptor with the antigen/MHC expressed on an antigen-presenting cell, T cell activation also requires costimulation by a second set of signals. The best characterized costimulatory receptor is CD28, which binds to a family of B7 ligands expressed on antigen-presenting cells. In asthma, although activated T cells play a role in the initiation and maintenance of airway inflammation, the importance of T cell costimulation in bronchial hyperresponsiveness had not been characterized. Therefore, we tested the hypothesis that inhibition of the CD28:B7 costimulatory pathway would abrogate airway hyperresponsiveness. Our results show that blockade of costimulation with CTLA4-Ig, a fusion protein known to prevent costimulation by blocking CD28:B7 interactions, inhibits airway hyperresponsiveness, inflammatory infiltration, expansion of thoracic lymphocytes, and allergen-specific responsiveness of thoracic T cells in this murine model of allergic asthma.

Topics: Airway Resistance; Animals; Bronchoalveolar Lavage; Bronchoconstrictor Agents; CD28 Antigens; Cell Division; Disease Models, Animal; Flow Cytometry; Histocytochemistry; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunohistochemistry; Inflammation; Lung; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes

We studied the function of autoinhibitory muscarinic M2 receptors on vagal nerve endings in the airways of conscious, unrestrained, ovalbumin-sensitized guinea pigs after the early and late allergic reaction. For this purpose, the effects of the selective muscarinic M2 receptor antagonist gallamine were examined on unilateral vagus nerve stimulation-induced bronchoconstriction, which was determined as an increase in basal respiration amplitude, measured as changes in pleural pressure. Under control conditions, i.e., before antigen challenge, a significant increase in the pleural pressure was found after inhalation of 0.1 mM and, even more pronounced, 1.0 mM gallamine, at medium stimulation frequencies (2-16 Hz), leading to a leftward shift of the frequency-response curve. After inhalation of 10 mM of gallamine, a complete reversal of the left-shift was observed and the frequency-response curve was depressed. However, 6 h after challenge with ovalbumin (i.e., after the early allergic reaction) no increase in nerve stimulation-induced bronchoconstriction by gallamine was found; a decrease in this bronchoconstriction was again observed with the highest concentration. At this moment, bronchial responsiveness to histamine was enhanced 4.5-fold compared to control, i.e., prior to antigen provocation. Both after the late allergic response (24 h after challenge; 1.6-fold histamine hyperresponsiveness) and 4 days after allergen challenge (normal histamine responsiveness) the gallamine-induced potentiation of the bronchoconstriction was restored, similar to the responses under control conditions. The results clearly demonstrate that prejunctional muscarinic M2 receptors control bronchoconstriction in conscious, unrestrained guinea pigs in vivo. Furthermore, these autoinhibitory receptors appear to be completely dysfunctional after the early allergic phase, but their function is largely restored after the late phase. The results indicate that dysfunction of autoinhibitory muscarinic M2 receptors might contribute to the strongly enhanced responsiveness to histamine after the early allergic response.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Electric Stimulation; Female; Gallamine Triethiodide; Guinea Pigs; Histamine; Hypersensitivity; Male; Ovalbumin; Receptor, Muscarinic M2; Receptors, Muscarinic; Vagus Nerve

A novel immunologically provoked inflammatory process was studied in guinea pigs. The animals were immunized by i.p. injections of ovalbumin (OA) suspended in Freund's complete adjuvant and challenged by the application of OA into the conjunctival sac of one eye. An inflammatory reaction was seen a few minutes after provocation and lasted normally for 4-7 days. The process was characterized by early damage to the epithelial layer which was partly detached in small flakes; an intense tearing with the tear fluid soon turning mucous and then purulent; vasodilation in the bulbar conjunctiva, in particular towards the limbal region; margination and emigration of polymorphonuclear, and to a lesser extent, eosinophil, leucocytes which migrated towards and infiltrated the surface epithelial layer. Subsequently, the dominant cell type infiltrating the submucosa was lymphocytes. Later, opacity of the cornea occurred, probably due to oedema and neovascularization of the stroma progressing centrally from the periphery. When the antigenic challenge was repeated, thickening of the conjunctival mucosa, and neoformation of collagen bundles in the submucosa led to the swelling of the upper lids. The facets of this inflammatory trauma may not fit easily into any of the classical types of hypersensitivity. Rather, it may combine features of several of them, at least type 1 and type 4. This syndrome shows several features similar to those of human vernal keratoconjunctivitis.

Topics: Animals; Conjunctiva; Conjunctivitis; Cornea; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Immunization; Leukocyte Count; Male; Microscopy, Electron; Ovalbumin

The effect of a thromboxane A2 (TXA2) receptor antagonist, ON-579, on experimental allergic skin and airway reactions was studied in vivo. ON-579 at doses of 1 and 20 mg/kg clearly inhibited U-46619-induced increases in respiratory resistance (Rrs) in guinea pigs. ON-579 at doses of 1, 20 and 50 mg/kg inhibited the aerosolized antigen-induced biphasic increase in Rrs in guinea pigs. Moreover, ON-579 clearly inhibited repeated aeroantigen-induced airway hyperreactivity in guinea pigs. ON-579, however, did not have any significant effects on allergic cutaneous reactions in rats. These results suggest that ON-579 is a relatively selective TXA2 antagonist, especially in the airways, and indicate the efficacy of ON-579 on antigen-induced increase in airway resistance and antigen-induced airway hyperreactivity in guinea pigs.

Topics: Aerosols; Airway Resistance; Animals; Antigens; Arthus Reaction; Bronchoalveolar Lavage Fluid; Concanavalin A; Dinitrophenols; Guinea Pigs; Histamine; Hypersensitivity; Ketotifen; Leukocyte Count; Male; Mice; Ovalbumin; Passive Cutaneous Anaphylaxis; Phenoxyacetates; Rats; Receptors, Thromboxane; Respiratory Tract Diseases; Serum Albumin, Bovine; Skin Diseases; Sulfonamides

The antiallergic constituents of oolong tea stem were examined. The stem extracts inhibited the 48 h homologous passive cutaneous anaphylaxis (PCA) reactions or rats in a dose-dependent manner and showed the same extent of inhibitory activity as ketotifen. All antiallergic constituents from the stem were concentrated into chloroform and ethyl acetate fractions, when extracted by various solvents. These fractions were treated with polyvinylpolypyrrolidone (PVPP), which resulted in the elimination of antiallergic activity in the ethyl acetate fraction, suggesting that one of the antiallergic constituents may be tea catechins. Then, six kinds of catechins, (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), -epicatechin (EC), (+)-catechin (C) and (-)-gallocatechin gallate (GCG), were isolated from the ethyl acetate fraction, and the inhibitory activity of these catechins on histamine release from rat peritoneal mast cells passively sensitized with anti-egg albumin (EA) IgE antibody was investigated. Among these catechins, significant inhibitory activity was observed in all the catechins except for EC. In addition, the inhibitory activity of GCG was greater than that of EGCG, which is well known to be an antiallergic constituent in tea. These results suggest that GCG may be a novel antiallergic constituent among tea catechins, and also the most potent.

Topics: Animals; Catechin; Histamine Release; Hypersensitivity; In Vitro Techniques; Mast Cells; Ovalbumin; Passive Cutaneous Anaphylaxis; Plant Extracts; Plant Stems; Rats; Rats, Wistar; Tea

The effects of two new immunosuppressors, FK-506 and mizoribine, on antigen-induced bronchial inflammation and reactivity to acetylcholine in mice were studied in comparison with those of cyclosporin A and cyclophosphamide. Three inhalations of an antigen by actively sensitized BALB/c mice resulted in an increase in airway reactivity to acetylcholine. Twenty-four hours after the final inhalation, the number of leukocytes (mononuclear cells and eosinophils) and the amount of interleukin 5 (IL-5) increased significantly. In BALB/c nu/nu mice (athymic mice), three inhalations of antigen caused no significant change in either airway inflammation or hyperresponsiveness. The administration of each of the four immunosuppressors clearly inhibited antigen-induced airway eosinophilia. Moreover, FK-506, mizoribine and cyclophosphamide clearly inhibited the antigen-induced IL-5 production and cyclosporin A showed the tendency to inhibit IL-5 production. Whereas FK-506, mizoribine and cyclosporin A clearly inhibited the antigen-induced airway hyperreactivity in BALB/c mice, cyclophosphamide did not show a significant effect on this airway hyperreactivity. These results indicate that FK-506, mizoribine and cyclosporin A, but not cyclophosphamide, inhibit antigen-induced airway hyperreactivity in mice. The mechanism which inhibits antigen-induced airway eosinophilia and IL-5 production is not involved in the inhibitory mechanism of airway hyperreactivity by FK-506 and mizoribine.

Topics: Aerosols; Animals; Bronchial Provocation Tests; Chemotaxis, Leukocyte; Cyclophosphamide; Cyclosporine; Hypersensitivity; Immunosuppressive Agents; Interleukin-5; Mice; Mice, Inbred BALB C; Mice, Nude; Ovalbumin; Respiratory System; Ribonucleosides; Tacrolimus

Interleukin-5 (IL-5) transgenic mice constitutively produce IL-5 and show massive eosinophilia in peripheral blood as well as infiltration of eosinophils into various tissues. This study investigated whether airway hyperreactivity occurs in the airways of IL-5 transgenic mice. These mice showed marked eosinophilia in bronchoalveolar lavage fluid, but no increase in reactivity to acetylcholine. IL-5 transgenic mice actively sensitized with ovalbumin displayed more profound airway hyperreactivity in response to acetylcholine 24 h after inhalation challenge with ovalbumin. Wild-type mice treated the same way showed little, if any, airway hyperreactivity. These findings indicate that this mouse model of airway hyperreactivity is useful in understanding the mechanism underlying pulmonary eosinophilia and the consequent inflammation.

Topics: Acetylcholine; Airway Resistance; Allergens; Animals; Bronchial Provocation Tests; Female; Hypersensitivity; Interleukin-5; Mice; Mice, Inbred C3H; Mice, Transgenic; Ovalbumin

1. We investigated whether virus-induced airway hyperresponsiveness in guinea-pigs could be modulated by pretreatment with capsaicin and whether viral respiratory infections could potentiate ovalbumin-aerosol-induced tracheal hyperresponsiveness. 2. Animals were inoculated intratracheally with bovine parainfluenza-3 virus or control medium 7 days after treatment with capsaicin (50 mg kg-1, s.c.). Four days after inoculation, tracheal contractions were measured to increasing concentrations of substance P, histamine and the cholinoceptor agonist, arecoline. 3. In tracheae from virus-infected guinea-pigs, contractions in response to substance P, histamine and arecoline were significantly enhanced (P < 0.01) by 144%, 46% and 77%, respectively. Capsaicin pretreatment inhibited the hyperresponsiveness to substance P partly (62%) and to histamine and arecoline completely. 4. In another series of experiments animals were first sensitized with ovalbumin (20 mg kg-1, i.p.). After 14 days animals were exposed to either saline or ovalbumin aerosols for 8 days. After 4 aerosol exposures (4 days) animals were inoculated with either parainfluenza-3 virus or control medium. One day after the last ovalbumin aerosol, tracheal contraction in response to increasing concentrations of substance P, histamine and arecoline was measured. 5. Tracheae from ovalbumin-aerosol-exposed control inoculated animals showed a similar degree of airway hyperresponsiveness to saline-aerosol-exposed virus-treated guinea-pigs. Virus inoculation of ovalbumin-treated animals significantly potentiated the tracheal contractions to substance P compared to either of the treatments alone. The contractions in response to histamine and arecoline were only slightly enhanced. 6. In conclusion, sensory nerves and/or tachykinins are involved in virus-induced airway hyperresponsivenessin guinea-pigs and viral respiratory infections can potentiate the increase in tracheal responsiveness to bronchoconstrictor agonists after ovalbumin exposure.

Topics: Animals; Arecoline; Capsaicin; Dose-Response Relationship, Drug; Guinea Pigs; Histamine; Hypersensitivity; Male; Ovalbumin; Respiratory Tract Infections; Substance P; Tachykinins; Trachea; Virus Cultivation

The ovalbumin-sensitized guinea pig is commonly used as a small animal model of allergic asthma. This animal model exhibits many of the hallmark characteristics observed in patients afflicted with asthma including nonspecific airway hyperreactivity, airway eosinophilia, early and late phase bronchoconstriction, and plasma extravasation into the airways. In addition, mucous hypersecretion in the airways of asthmatic patients is thought to be responsible for the plugging of distal airways and to contribute to the morbidity and mortality associated with the disease process. In this study we examined whether the allergic guinea pig model exhibits an increase in airway high molecular weight glycoconjugate (HMWG) secretion in response to an antigen challenge and whether dexamethasone exerts any modulatory effects upon the response. Ovalbumin (OVA) -sensitized guinea pigs were challenged with OVA 2 wk following the initial exposure. Trachobronchoalveolar lavages (TBAL) were performed, and the samples were assayed for total eosinophil cell number, eosinophil peroxidase activity (EPO), and both acidic and neutral HMWG content. Morphometric analysis of mucous-containing cells was also performed on tissue sections prepared from the trachea, mainstem bronchus, and three lobes of the left lung. Within 24 h of an antigen challenge, TBAL samples obtained from the allergic guinea pigs exhibited increases in eosinophil cell number, measured EPO enzyme activity, and acidic HMWG content compared to TBAL samples prepared from vehicle-exposed animals. These antigen-induced changes were dependent on the concentration of aerosolized OVA administered. Exposing the animals to 0.3% OVA provoked a 6.23-fold increase in airway eosinophils, 15-fold elevation in TBAL EPO enzyme activity, and 175% increase in TBAL acidic HMWG. No significant changes in TBAL neutral HMWG were measured. The changes in measured EPO activity correlated with the levels of acidic HMWG found in the TBAL samples (r = 0.73, P < or = 0.001). The measured increase in TBAL acidic HMWG was time dependent and was found to be maximal at 2 h post-antigen challenge. Morphometric analysis of Alcian blue (pH 2.5) -stained airway sections showed a decline in stored mucosubstances following the antigen exposure, supporting the notion that the allergic guinea pig model exhibits a mucosecretory component. Pretreating the animals with dexamethasone attenuated the antigen-induced release of HMWG and changes in measured EPO act

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Dexamethasone; Disease Models, Animal; Eosinophils; Glycoconjugates; Guinea Pigs; Hypersensitivity; Male; Molecular Weight; Mucus; Ovalbumin

Activated CD4+ helper T cells have been demonstrated in asthmatic airways and postulated to play a central role in eliciting allergic inflammation; direct evidence of their involvement seems to be lacking. We hypothesized that CD4+ T cells have the potential to induce allergic responses to antigen challenge, and tested this hypothesis in a model of allergic bronchoconstriction, the Brown Norway rat, using the approach of adoptive transfer. Animals were actively sensitized to either ovalbumin (OVA) or BSA and were used as donors of T cells. W3/25(CD4)+ or OX8(CD8)+ T cells were isolated from the cervical lymph nodes of sensitized donors and transferred to naive BN rats. 2 d after adoptive transfer recipient rats were challenged by OVA inhalation, and changes in lung resistance (RL), bronchoalveolar lavage (BAL) cells, and serum levels of antigen-specific IgE were studied. After OVA challenge recipients of OVA-primed W3/25+ T cells exhibited sustained increases in RL throughout the entire 8-h observation period and had significant bronchoalveolar lavage eosinophilia, which was detected by immunocytochemistry using an antimajor basic protein mAb. Recipients of BSA-primed W3/25+ T cells or OVA-primed OX8+ T cells failed to respond to inhaled OVA. OVA-specific immunoglobulin E was undetectable by ELISA or skin testing in any of the recipient rats after adoptive transfer. In conclusion, antigen-induced airway bronchoconstriction and eosinophilia were successfully transferred by antigen-specific W3/25+ T cells in Brown Norway rats. These responses were dependent on antigen-primed W3/25+ T cells and appeared to be independent of IgE-mediated mast cell activation. This study provides clear evidence for T cell mediated immune mechanisms in allergic airway responses in this experimental model.

Topics: Allergens; Animals; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Eosinophils; Hypersensitivity; Immunity, Cellular; Immunoglobulin E; Immunotherapy, Adoptive; Inflammation; Leukocyte Count; Lung; Lymph Nodes; Lymphocyte Activation; Lymphocyte Transfusion; Male; Ovalbumin; Rats; Rats, Inbred BN; Respiratory System; Serum Albumin, Bovine; Skin Tests; T-Lymphocyte Subsets

Mast cells are important effector cells in IgE-mediated acute allergic reactions. Mast cells also produce cytokines such as interleukin (IL)-3, IL-4, IL-5, tumor necrosis factor (TNF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) that regulate the function of eosinophils and the development of a late-phase inflammatory response to antigen challenge. To evaluate the role of mast cells on the development of IgE-mediated allergic pulmonary eosinophilia in vivo, we compared the eosinophil infiltration into lungs of mast cell deficient mice (WBB6F1/J-W/Wv) with their congenic normal littermates (W/W+). Mice were sensitized with alum-precipitated ovalbumin and challenged with aerosolized ovalbumin on day 12 after sensitization. Bronchoalveolar lavage (BAL) fluid, lung tissue biopsies, and blood samples were collected after ovalbumin challenge. Eosinophil numbers in the BAL and lung tissue, lung eosinophil peroxidase (EPO) activity and serum levels of IgE and IgG1 were measured. In sensitized W/W+ mice, there were increased numbers of eosinophils in the BAL fluid and lung tissue, and EPO levels were increased after ovalbumin challenge. Ovalbumin challenge of sensitized mast-cell-deficient mice produced fewer numbers of eosinophils in the BAL fluid and lungs, and EPO levels were also reduced compared with their challenged congenic littermates. On the other hand, levels of serum IgE and IgG1 were not different between W/Wv mice and their congenic littermates.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Bone Marrow Cells; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Cell Movement; Eosinophil Peroxidase; Eosinophils; Hypersensitivity; Immunization, Passive; Immunoglobulin E; Immunoglobulin G; Male; Mast Cells; Mice; Mice, Mutant Strains; Ovalbumin; Peroxidases; Pneumonia

The effect of an Ascaris suum extract (Asc) on several T cell functions was studied in mice immunized with Asc and ovalbumin (OA) in complete Freund's adjuvant. Delayed-type hypersensitivity reactions following challenge with aggregated OA were markedly diminished in mice injected with OA plus a high dose of Asc compared to OA-immunized animals. Proliferation and IL-2, IFN-gamma, IL-4, and IL-10 production in OA-stimulated lymph node cells from the OA + Asc-immunized group were also inhibited. Titration of anti-OA antibodies also showed suppression of IgG1, IgG2a, and IgE isotypes in the animals injected simultaneously with the extract. The degree of suppression induced by Asc on OA-specific cell-mediated responses was dose dependent. The profile of cytokines synthesized in response to Asc also changed depending on the injected dose. IL-4 and IL-10 were mainly produced in response to high doses, whereas IL-2 and IFN-gamma were greatly enhanced at a low dose of Asc. These findings indicate that the A. suum extract may impair crucial T cell functions for cell-mediated as well as humoral immune responses to other antigens through the induction of a predominantly Th2-like response.

Topics: Animals; Antibodies, Helminth; Antibody Formation; Antigens, Helminth; Ascaris suum; Dose-Response Relationship, Immunologic; Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulin Isotypes; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Lymphocyte Activation; Male; Mice; Mice, Inbred DBA; Ovalbumin; T-Lymphocytes

Inflammatory challenges of the airway mucosa cause luminal entry of bulk plasma. Extravasation of plasma is well described but the routes for epithelial passage of plasma are largely unknown. Using colloidal gold (5 nm) as tracer we have now examined the fate of extravasated plasma in the airways. The tracer was given intravenously to anaesthetized, ovalbumin-sensitized guinea-pigs 2 min prior to airway mucosal challenge with 12 pmol ovalbumin (the dose was selected from a separate dose-response study). Tissue specimens were collected 30 s, 3 and 6 min after end of challenge (separate time course experiments suggested that the peak rate of entry of plasma occurred at about 5 min). The colloidal gold particles were visualized by autometallographic silver intensification. The gold produced no circulatory disturbance and had a uniform vascular distribution with negligible adherence to vascular endothelium. After challenge gold was first widely distributed in the lamina propria. At 3 and 6 min the tracer was also in the epithelium and airway lumen. It appeared that plasma was moved distinctly between and all around each epithelial cell. Bright field-, scanning-, and transmission electron-microscopy indicated that the luminal entry of plasma did not affect the integrity of the epithelial lining. This study demonstrates that the plasma exudate moves across an intact epithelial layer through ubiquitous paracellular pathways.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Allergens; Animals; Biological Transport; Dose-Response Relationship, Drug; Epithelium; Exudates and Transudates; Gold; Guinea Pigs; Hypersensitivity; Injections, Intravenous; Laryngeal Mucosa; Male; Microscopy, Electron; Ovalbumin; Time Factors

This report examines the effect of recombinant murine (rm) IL-10 on antigen-induced cellular recruitment into the airways of sensitized Balb/c mice. The intranasal instillation of 10 micrograms ovalbumin induced an early (6-24 h) increase in the number of neutrophils, and a late rise (24-96 h) in that of eosinophils in the bronchoalveolar lavage (BAL) fluid and bronchial tissue. A single intranasal instillation of 0.01-0.1 microgram of rmIL-10, administered concurrently with ovalbumin, but not 1 or 3 h thereafter, dose-dependently inhibited both airway neutrophilia and eosinophilia. This phenomenon was suppressed by treating the sensitized mice with 1 mg/mouse of a neutralizing anti-IL-10 mAb, which increased significantly ovalbumin-induced neutrophil and eosinophil accumulation in the BAL fluid. These results suggest that antigen stimulation may trigger the in vivo generation of IL-10, which, in turn, participates in the leukocyte infiltration into the airways. rmIL-10 also reduced TNF-alpha release in the BAL fluid observed 1 and 3 h after antigen challenge. Furthermore, the intranasal instillation of an anti-TNF-alpha antiserum to sensitized mice markedly reduced ovalbumin-induced neutrophil and eosinophil accumulation in the BAL fluid. These findings indicate that leukocyte infiltration into the airways of antigen-challenged mice is regulated by IL-10. Furthermore, inhibition of TNF-alpha production by rmIL-10 suggests that allergic airway inflammation and TNF-alpha formation are parallel events in this model.

Topics: Administration, Intranasal; Animals; Antigens; Bronchi; Bronchoalveolar Lavage Fluid; Hypersensitivity; Interleukin-10; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Proteins; Tumor Necrosis Factor-alpha

A new guinea pig model of allergic asthma was used to investigate the effects of low doses of the phosphodiesterase inhibitors, rolipram (phosphodiesterase IV selective), ORG 20241 (N-hydroxy-4-(3,4-dimethoxyphenyl)-thiazole-2-carboximidamide; dual phosphodiesterase III/IV inhibitor with some selectivity for the phosphodiesterase IV isoenzyme), and of theophylline (non-selective) on allergen-induced early and late phase asthmatic reactions, bronchial hyperreactivity to histamine inhalation, and airway inflammation. Theophylline (25 mg/kg i.p.) and ORG 20241 (5 mg/kg i.p.) did not affect histamine-induced bronchoconstriction, whereas rolipram (75 micrograms/kg i.p.) only slightly reduced the response to histamine at 7 h after administration. However, bronchial hyperreactivity after the early and after the late reaction was significantly reduced by theophylline, rolipram and ORG 20241, while bronchoalveolar lavage studies revealed a selective inhibition of airway inflammation by the phosphodiesterase inhibitors. Theophylline significantly reduced the number of eosinophils, neutrophils and macrophages; rolipram reduced the number of neutrophils and lymphocytes, and ORG 20241, the number of eosinophils and macrophages. None of the compounds at the dosage indicated reduced the early and late reaction when administered i.p. 1 h before allergen exposure to defined dual responding animals. These results indicate that non-bronchodilator doses of these phosphodiesterase inhibitors markedly reduce the allergen-induced development of bronchial hyperreactivity as well as airway inflammation, presumably by selectively inhibiting cellular migration. The results suggest that an orchestrated series of cellular interactions is involved in the development of bronchial hyperreactivity. It is concluded that phosphodiesterase inhibitors may be very useful in the treatment of bronchial asthma.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Administration, Inhalation; Analysis of Variance; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cyclic Nucleotide Phosphodiesterases, Type 4; Disease Models, Animal; Eosinophils; Guinea Pigs; Histamine; Hypersensitivity; Inflammation; Injections, Intraperitoneal; Isoenzymes; Macrophages; Male; Neutrophils; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Pyrrolidinones; Rolipram; Specific Pathogen-Free Organisms; Theophylline; Thiazoles

T helper 2 (Th2)-like cytokines are thought to play a crucial role in the pathogenesis of airway inflammation in atopic asthma, leading to bronchial hyperresponsiveness. To investigate the role of the principal Th2 cytokine interleukin-4 (IL-4) in asthma, we examined the allergen-induced changes in airway morphology and bronchial responsiveness (BR) in an in vivo mouse model. C57BL/6 mice were actively sensitized to ovalbumin (OVA) and exposed daily to aerosolized OVA or saline (SAL) for 7 days. Twenty-four hours after the last allergen exposure, total and differential counts of bronchoalveolar lavage cells revealed a significant increase of eosinophils and lymphocytes in OVA-exposed immunized mice compared with SAL-exposed animals. In IL-4-deficient (IL-4-/-) mice, treated in the same way, there were substantially fewer eosinophils in bronchoalveolar lavage compared with wild-type mice. Allergen exposure of actively sensitized wild-type mice induced a significant increase of BR to carbachol and to serotonin compared with SAL-exposed mice. In contrast, OVA exposure of immunized IL-4-/- mice did not augment BR to serotonin compared with SAL-challenged IL-4-/- mice. In conclusion, these data indicate that repeated allergen exposure in sensitized mice induces airway inflammation and bronchial hyperresponsiveness, and that IL-4 plays a predominant role in the pathogenesis of both phenomena.

Topics: Allergens; Animals; Asthma; Bronchial Provocation Tests; Carbachol; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Serotonin

Topics: Animals; Bone Marrow; Hypersensitivity; Immunotherapy; Interferon-gamma; Interferons; Interleukin-4; Interleukin-5; Mice; Mice, Inbred Strains; Ovalbumin; Pulmonary Eosinophilia; Recombinant Proteins

The possible involvement of potassium (K) channels in allergic airway responses was examined in ovalbumin (OA)-sensitized guinea pigs. An ATP-sensitive K channel opener (BRL-38227) inhibited OA inhalation-induced bronchoconstriction and airway plasma leakage. BRL-38227 also had an inhibitory effect on exogenous histamine- and leukotriene-induced responses. In contrast, BRL-38227 did not affect OA-induced histamine release from minced lung tissues. Therefore, the ATP-sensitive K channel opener inhibits allergic bronchoconstriction and plasma leakage as a result of its effect on airway smooth muscle and postcapillary venules. Apamin, a small conductance Ca2+ -activated K channel (PK,Ca) blocker, significantly inhibited both OA-induced tracheal contraction and histamine release from lung tissues, suggesting that this compound reduces allergic airway responses via a mast cell stabilizing effect. We conclude that ATP-sensitive K channel opening and small conductance PK,Ca closure may be beneficial for preventing allergic airway responses.

Topics: Adenosine Triphosphate; Animals; Benzopyrans; Bronchoconstriction; Capillary Permeability; Cromakalim; Guinea Pigs; Histamine Release; Hypersensitivity; Male; Ovalbumin; Potassium Channels; Pyrroles

Topics: Animals; CD4-Positive T-Lymphocytes; Eosinophils; Hypersensitivity; Interferon-gamma; Lymphocyte Depletion; Male; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Peritoneal Cavity; Peritonitis; Recombinant Proteins; Tacrolimus

Airway hyperreactivity to physical, chemical, immunological and pharmacological stimuli is well documented in vivo. The aim of this study was to investigate whether tissues taken from guinea-pigs that had been shown to display hyperreactivity in vivo after antigen challenge were also hyperreactive in vitro. Isolated airway-perfused lungs from ovalbumin-sensitized guinea-pigs challenged 24 h beforehand with an aerosol of ovalbumin showed a significant (P < 0.05) increase in responsiveness to the bronchoconstrictor response to a bolus dose of carbachol (10 micrograms) when compared with saline challenged animals. The contractile responses to single doses of carbachol (10 micrograms) and histamine (30 micrograms) in immersed tracheal spiral preparations taken from sensitized animals exposed to the ovalbumin were also significantly enhanced (P < 0.05). A non-significant leftward shift was observed in the concentration-response curve for histamine in challenged perfused lungs from ovalbumin-challenged animals compared with an NaCl challenge. Concentration-response curves to carbachol and histamine in immersed tracheal spirals were virtually superimposed. Therefore, this study has shown non-specific airway hyperreactivity of isolated airway perfused lungs at 24 h following a challenge of sensitized guinea-pigs with aerosolized ovalbumin, although this was not evident from concentration-response curves in immersed trachea. The isolated perfused lung therefore provides a simple method for further evaluation of the mechanisms of airway hyperreactivity.

Topics: Adenosine; Animals; Carbachol; Dose-Response Relationship, Drug; Guinea Pigs; Histamine; Hypersensitivity; Lung; Male; Muscle, Smooth; Ovalbumin; Respiratory System; Trachea

We have recently developed synthetic low molecular weight inhibitors of both tissue and plasma kallikreins. Several of these were evaluated in vivo in the ovalbumin-sensitised guinea pig for their ability to prevent the bronchoconstriction elicited by antigen challenge. The selective tissue kallikrein inhibitor CH-694 (but not the selective plasma kallikrein inhibitor CH-684) caused highly significant falls in airways resistance when it was administered at 10 mg/kg intraperitoneally 15 min before and 90 min after challenge. There was also a highly significant fall in the tissue kallikrein activity measured in broncho-alveolar lavage fluid. Inhibitors of tissue kallikrein may prove effective in the treatment of allergic inflammation in man.

Topics: Airway Resistance; Aldehydes; Amino Acid Sequence; Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Inflammation; Kallikreins; Ketones; Molecular Sequence Data; Ovalbumin; Peptides; Tissue Kallikreins

Relaxant effects of the beta 2-selective adrenoceptor agonist TA-2005 on bronchoconstriction in the anesthetized guinea pig and cat were evaluated in comparison with other known beta 2-adrenoceptor agonists. The ED50 values of intravenously administered TA-2005, procaterol, formoterol, isoproterenol, salbutamol, and salmeterol to inhibit the histamine-induced bronchoconstriction of the guinea pigs were 0.024, 0.053, 0.056, 0.099, 0.23, and 2.00 micrograms/kg, respectively, and those in serotonin-challenged cats were 0.019, 0.037, 0.039, 0.042, 0.13, and 0.52 micrograms/kg, respectively, in the same increasing order. When guinea pigs were passively sensitized with anti-ovalbumin antiserum, the ED50 values of TA-2005, formoterol, procaterol, and isoproterenol to inhibit the antigen-induced bronchoconstriction were 0.09, 0.30, 0.65, and 7.0 micrograms/kg, i.v., respectively, while those of TA-2005, procaterol, formoterol, and salbutamol in actively sensitized animals were 0.24, 0.25, 1.40, and 23.0 micrograms/kg. When TA-2005 was administered by inhalation to guinea pigs or by the intraduodenal route to cats, it exhibited a long-lasting inhibitory effect comparable or superior to the effects of salmeterol and formoterol. These data indicate that, among the known beta 2-adrenoceptor agonists examined, TA-2005 exerts the most potent bronchodilating effects with a long duration of action in vivo, and its potency ratios to the other reference drugs were greater in antigen- than spasmogen-induced bronchoconstriction models.

Topics: Administration, Inhalation; Administration, Oral; Adrenergic beta-2 Receptor Agonists; Adrenergic beta-Agonists; Amphetamines; Animals; Bronchodilator Agents; Cats; Dose-Response Relationship, Drug; Female; Guinea Pigs; Histamine Antagonists; Hydroxyquinolines; Hypersensitivity; Injections, Intravenous; Male; Myocardial Contraction; Ovalbumin; Quinolones; Respiratory Function Tests; Serotonin

Proliferative response of cord blood lymphocytes stimulated twice by food antigens and cord blood IgE concentration were measured in 131 full-term newborn infants for the prediction of allergic disorders. Through the follow-up study for 2 1/2 years, the value of stimulation index in proliferative response of cord blood lymphocytes to ovalbumin or bovine serum albumin was greater than 1.5 in 17 (sensitivity 53.1%) of 32 infants in whom allergic disorders developed and less than 1.5 in 81 (specificity 81.8%) of 99 infants who had no allergic disorders (cutoff limit of stimulation index 1.5). The sensitivity was increased (71.9%) by the combination of the cord blood IgE concentration (cutoff limit 1.0 IU/ml) and proliferative response of cord blood lymphocytes to food antigens (cutoff limit of stimulation index 1.5). The combination of the cord blood IgE concentration and proliferative response of cord blood lymphocytes to food antigens is useful for the prediction of allergic disorders. Interleukin-2 production of cord blood lymphocytes stimulated with food antigens was also measured in 24 newborn infants. Interleukin-2 activity in culture supernatants of ovalbumin- or bovine serum albumin-stimulated cord blood lymphocytes correlated well with proliferative response of cord blood lymphocytes to ovalbumin or bovine serum albumin.

Topics: Antigens; Blood Cells; Cell Division; Cells, Cultured; Fetal Blood; Follow-Up Studies; Food; Humans; Hypersensitivity; Immunoglobulin E; Infant, Newborn; Interleukin-2; Lymphocytes; Osmolar Concentration; Ovalbumin; Predictive Value of Tests; Serum Albumin, Bovine

We previously reported that intestine from rats sensitized to ovalbumin (Ova), using Bordetella pertussis vaccine as adjuvant, demonstrated a rapid secretory response [increase in short-circuit current (Isc)] to Ova upon secondary challenge. Here, we examined the role of pertussis toxin, the active component of the vaccine, in the response. Sensitization of Sprague-Dawley rats by intraperitoneal injection of recombinant wild-type pertussis toxin (wPT) plus Ova enhanced intestinal responses (at day 14: approximately 20-fold for luminal antigen, approximately 2.5-fold for serosal antigen) compared with rats sensitized by injection of Ova alone. In contrast, sensitization with an enzymatically inactive mutant pertussis toxin (mPT, different in two amino acids) produced no significant effect. Ova-specific immunoglobulin (Ig) E and IgG2a antibodies and greater numbers of mucosal mast cells were documented in wPT-sensitized rats. In addition, the Isc response to electrical transmural stimulation of nerves in intestinal preparations was significantly augmented. Neurotoxin inhibited the secretory response to luminal but not serosal antigen. Immunophysiological stimulation by wPT was still evident 8 mo postsensitization. Our studies indicate that pertussis toxin causes long-lasting hypersensitivity to coadministered antigens, involving increased production of reaginic antibodies, hyperplasia of mucosal mast cells, and enhanced neurally mediated uptake of antigen across the intestinal epithelium. These findings suggest a potential role for bacterial products in the development of immunophysiological reactions to ingested antigens.

Topics: Animals; Antibodies; Antigens; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Intestinal Mucosa; Intestines; Male; Mast Cells; Mutation; Nervous System Physiological Phenomena; Ovalbumin; Pertussis Toxin; Rats; Rats, Sprague-Dawley; Transcutaneous Electric Nerve Stimulation; Virulence Factors, Bordetella

Many attempts have failed to correlate in vivo airway responsiveness with in vitro airway smooth muscle functions. We have reexamined this relation by taking account of airway epithelial functions in guinea pigs sensitized with inhaled ovalbumin (OA). In vivo responses were assessed by the provocative concentration of acetylcholine (ACh) required to double the airway opening pressure (PC200) under mechanical ventilation. In vitro responses were measured in a perfused whole-tracheal preparation. The negative logarithm of the molar concentration of ACh required to produce a 10% reduction in diameter was calculated both for epithelial-side stimulation (PC10(in)) and for serosal-side stimulation (PC10(out)). OA-sensitized guinea pigs showed significantly smaller log PC200 than control animals (0.51 +/- 0.07 and 0.81 +/- 0.10, respectively, p < 0.01). In in vitro study, there were variable differences in PC10(in) and PC10(out) in each animal. The difference in sensitivity between epithelial- and serosal-side stimulation (PC10(in-out)) showed a significant correlation in PC10(in) (r = 0.82, n = 9, p < 0.01) but not to PC10(out) (r = 0.39, p > 0.1), indicating that the variation in PC10(in-out) resulted from the changes in PC10(in). For in vivo-in vitro correlation, log PC200 correlated significantly with PC10(in) (r = 0.68, n = 9, p < 0.05) but not with PC10(out) (r = 0.18, p > 0.1). These results indicate that the sensitization by inhalation of OA produces increased airway responsiveness to ACh in vivo and that this airway responsiveness may be related, at least in part, to the altered airway epithelial functions.

Topics: Acetylcholine; Administration, Inhalation; Airway Resistance; Animals; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Disease Models, Animal; Dose-Response Relationship, Drug; Epithelium; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Least-Squares Analysis; Male; Muscle, Smooth; Ovalbumin; Respiration, Artificial; Trachea

Cholera toxin (CTX) is a potent oral adjuvant for the induction of mucosal IgA Ab responses protein Ags. We examined the Ab responses and allergic sensitization of several strains of mice to protein Ags, administered orally with CTX. The mice made strong IgA and IgG1 serum Ab responses, but little IgG2a Ab to Ags such as hen egg lysozyme (HEL) and OVA. However, when given a subsequent i.p. challenge with Ag alone, the same mice had immediate hypersensitivity reactions that included respiratory distress and death. Within 10 min of i.p. challenge, immunized mice had high levels of plasma histamine and extensive degranulation of mast cells in target tissues. These mice had detectable serum IgE Ab. Ag administered orally with the B subunit (CTB) of CTX did not sensitize mice. Intestinal tissues taken from these mice had Ag-specific ion-secretory responses in vitro, typical of intestinal anaphylaxis. Ag given s.c. without adjuvant could also sensitize for systemic and intestinal anaphylaxis. Sensitization with HEL given s.c. was dose dependent and correlated with a critical amount of HEL in the circulation. HEL was detected in the circulation after oral immunization, but CTX did not increase the uptake of HEL. Thus, oral immunization with a protein Ag in the presence of CTX can sensitize an animal for systemic and intestinal anaphylaxis. These results suggest a cautious approach to the use of CTX as an adjuvant in oral vaccines, and provide a new model to study immediate hypersensitivity reactions to intestinal Ag.

Topics: Administration, Oral; Anaphylaxis; Animals; Antigens; Cholera Toxin; Female; Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Intestines; Mice; Mice, Inbred C3H; Muramidase; Ovalbumin

1. In guinea-pigs previously sensitized with ovalbumin, the intra-plantar administration of the antigen induced dose-dependent and sustained oedema. An intense infiltrate of neutrophils and eosinophils was observed at the peak of the oedema (4 h). 2. Oedema induced by ovalbumin at the doses of 50 or 200 micrograms/paw was not inhibited by antihistamines (meclizine and cetirizine), a PAF antagonist (BN 50730), a cyclo-oxygenase inhibitor (indomethacin), a lipoxygenase inhibitor (MK-886), a dual type lipo- and cyclo-oxygenase inhibitor (NDGA), a bradykinin antagonist (Hoe 140) or the combination of cetirizine, MK-886, indomethacin and BN 50730. These drugs did inhibit paw oedema induced by their specific agonists or by carrageenin. These results suggest that histamine, PAF, prostaglandins, leukotrienes or bradykinin are not important in the development of immune paw oedema in guinea-pigs. 3. Dexamethasone (10 mg kg-1) inhibited oedema induced by ovalbumin (50 or 200 micrograms/paw, P < 0.05). This effect apparently does not result from inhibition of arachidonate metabolism, since indomethacin, MK-886 and NDGA were without effect. 4. Oedema induced by ovalbumin (50 or 200 micrograms/paw) was also inhibited by azelastine. This effect was not due to the anti-histaminic property of azelastine since two other potent-antihistamines, meclizine and cetirizine, were ineffective. 5. Intravenous injection of lipopolysaccharide (LPS) dose-dependently inhibited the oedema induced by ovalbumin (200 micrograms/paw). This effect could not be attributed to hypotension or leucopenia since the maximal dose applied (81 micrograms kg-1) did not induce significant changes in the blood pressure or in the white blood cell levels of the animals. It is suggested that the effect of LPS is mediated by the endogenous release of cytokines, including tumour necrosis factor (TNF alpha). Murine TNF alpha dose dependently(9-81 microg kg-1) inhibited the paw oedema induced by ovalbumin.7. The anti-oedematogenic effects of LPS and/or TNF alpha are possibly associated with their capacity to inhibit leucocyte emigration. Accordingly, guinea-pigs rendered leucopenic with vinblastine exhibited less intense oedema after ovalbumin. Vinblastine did not affect oedema induced by PAF or bradykinin,indicating that vascular responsiveness was not involved.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Blood Pressure; Dexamethasone; Edema; Enzyme-Linked Immunosorbent Assay; Foot; Guinea Pigs; Hypersensitivity; Immunoglobulin G; Leukocyte Count; Leukocytes; Leukopenia; Lipopolysaccharides; Male; Mice; Ovalbumin; Phthalazines; Platelet Aggregation Inhibitors; Tumor Necrosis Factor-alpha; Vinblastine

Parasite-naive guinea pigs with genetically determined differences in responsiveness to infection with the gastrointestinal nematode parasite Trichostrongylus colubriformis were sensitised to ovalbumin and later challenged by exposure to an ovalbumin aerosol. The resultant cellular migration into the lungs was assessed by histological examination of the lungs and enumeration of cells in bronchoalveolar lavage fluid 24 h, 72 h and 7 days later. Compared with parasite-low-responder guinea pigs, there were approximately 10 times more eosinophils in lavage fluid from parasite-high-responder animals but similar numbers of neutrophils.

Topics: Aerosols; Animals; Antigens; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Eosinophilia; Eosinophils; Guinea Pigs; Hypersensitivity; Lung; Male; Neutrophils; Ovalbumin; Trichostrongylosis; Trichostrongylus

The functional role of CD8 T cells in in vivo IgE production, immediate cutaneous reactivity, and altered airways responsiveness (AR) was examined in a murine model of allergen-induced sensitization. Exposure of BALB/c mice to nebulized OVA triggered an IgE anti-OVA response in the serum, immediate-type skin test responses to OVA, and the development of increased AR (as measured by nonspecific reactivity to electrical field stimulation). In spleens of sensitized mice, analysis of the distribution of CD4/CD8 T cell subpopulations revealed an increase in total numbers of CD8 T cells. Transfer of purified spleen CD8 T cells from OVA-sensitized mice (CD8OVA) to sensitized recipients reduced serum IgE anti-OVA production by roughly 50%. Furthermore, studies of in vitro Ig production indicated that mononuclear cells from recipients of CD8 cells (CD8OVA > CD8PBS) produced less IgE and IgG1 antibodies, whereas in vitro IgG2a production was enhanced. The suppression of IgE production in recipients of CD8OVA T cells was associated with the failure to respond to intradermal challenge with OVA. The increase in AR found in sensitized mice was prevented after transfer of CD8OVA cells. When CD8 T cells from nonimmunized animals (CD8PBS) were used for the transfer into sensitized recipients, serum anti-OVA IgE was decreased by only 20%, whereas skin test reactivity and AR were not significantly affected. The ex vivo analysis of the pattern of cytokine-producing lymphocytes by immunofluorescence microscopy indicated that the sensitization procedure increased the fraction of IFN-gamma- and IL-4-positive cells in the spleen. Further in vitro analysis demonstrated that a high percentage of CD8 T cells were positive for IFN-gamma, whereas IL-4 was produced mainly by CD4 T cells. These data suggest that CD8 T cells may play an important role in the negative regulation of IgE production and AR and that IFN-gamma may be a relevant mediator of the functions of CD8 T cells in this model.

Topics: Animals; CD8 Antigens; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Skin Tests; Spleen; T-Lymphocytes, Regulatory

Using guinea pig models of nasal allergy, we investigated the role of PAF in mucosal hypersensitivity to topical histamine challenge. We found that the histamine sensitivity was significantly increased by nasal allergen challenge in actively sensitized guinea pigs, and the occurrence of histamine hypersensitivity was inhibited by pretreatment of anti-PAF agent (SM10661, 10 mg/kg, i.p.). Also, the histamine sensitivity was further increased by a single nasal challenge with PAF in actively sensitized guinea pigs. The histamine hypersensitivity was induced by repeated topical challenges with PAF in nonsensitized guinea pigs. Therefore, we believe that PAF plays an important role in hypersensitivity to histamine in nasal allergy.

Topics: Allergens; Animals; Guinea Pigs; Histamine Release; Hypersensitivity; Male; Ovalbumin; Platelet Activating Factor; Thiazoles; Thiazolidines

A number of N-[4-[4-(1H-indol-3-yl)piperidinoalkyl]-2- thiazolyl]alkanesulfonamides (8--21) were synthesized and evaluated for their preventive effects on systemic anaphylaxis in guinea pigs. Structure-activity analysis revealed that methane- and ethanesulfonamide derivatives having a one to three methylene tether between the piperidine and thiazole rings exhibited potent activity but the introduction of a substituent on the indole part reduced the activity. Administration (100 mg/kg p.o.) of the four compounds 8, 9, 12, 13, together with ketotifen, oxatomide, terfenadine and azelastine as reference compounds, to mice revealed that only compound 8 caused no significant increase of the sleeping time induced by hexobarbital. In addition, compound 8 (10 mg/kg i.v.) did not change the electroencephalogram in conscious rabbits. These results led to the selection of N-[4-[4-(1H-indol-3-yl)piperidinomethyl]-2-thiazolyl]methanesulfon amide (8, FK613) for further development as a novel antiallergic agent. Clinical evaluation of FK613 is now in progress.

Topics: Anaphylaxis; Animals; Guinea Pigs; Hypersensitivity; Indoles; Male; Mice; Mice, Inbred ICR; Ovalbumin; Rabbits; Structure-Activity Relationship; Sulfonamides; Thiazoles

Recent evidence supports a role for T lymphocytes in allergic airway responses. We hypothesized that reducing blood T suppressor cells (Ts) might increase the late airway response (LR). Sprague-Dawley (SD) rats were sensitized with ovalbumin (OA). On days 8, 10, and 12, post-sensitization test SD (n = 14) received monoclonal antibody intravenously (OX-8; 1 mg) specific to rat Ts. Controls received saline (n = 7) or mouse ascites IgG (n = 7). On day 14, animals were challenged with OA aerosol (5% wt/vol) for 5 min, lung resistance was recorded for 8 h (n = 18) and bronchoalveolar lavage was performed. The LR was determined from the area under the lung resistance vs time curve from 75 to 480 min after challenge. In the remaining 10 rats, airway lymphocyte subsets were measured 8 h after OA aerosol challenge in minced and digested lungs. A decrease in percentage of blood and airway Ts, respectively, in test animals was observed vs controls (blood: 6.27 +/- 0.84 vs 32.95 +/- 1.94, P < 0.001); (airway: 5.05 +/- 0.66 vs 24.5 +/- 3.05, P < 0.02). Blood and airway helper T lymphocytes did not differ between test and control animals. The LR was significantly increased in test (22.89 +/- 3.92) vs controls (4.22 +/- 2.18, P < 0.001). Bronchoalveolar lavage macrophages, neutrophils and lymphocytes, and serum OA-specific IgE were also significantly elevated (P < 0.05) in test animals. We conclude that Ts play an important role in attenuating the LR in SD rats.

Topics: Aerosols; Animals; Bronchial Provocation Tests; Bronchial Spasm; Bronchoalveolar Lavage Fluid; Hypersensitivity; Immunoglobulin E; Lymphocyte Depletion; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Time Factors

Basal adenylyl cyclase activity in lung membranes isolated from hypersensitive guinea pigs was increased and more sensitive to stimulation by isoprenaline, GTP and GppNHp when compared to adenylyl cyclase in lung membranes isolated from normal healthy guinea pigs. Maximal forskolin-stimulated adenylyl cyclase activity was unaltered. There was no change in the immunological quantitative amounts of either alpha subunits of the G proteins GiII and Gs (G(o), GiI and GiIII were not present). Maximal pertussis-toxin- and cholera-toxin-catalyzed ADP-ribosylation of Gi alpha and Gs alpha respectively were not significantly altered. The addition of purified protein kinase C to isolated lung membranes resulted in the phosphorylation of the alpha subunit of Gs (stoichiometry was 0.53 mol of 32P incorporated/mol of Gs alpha). Addition of protein kinase C to lung membranes isolated from hypersensitive guinea pigs was equally effective at catalysing the phosphorylation of the alpha subunit of Gs. GppNHp-stimulated and basal adenylyl cyclase activity was also enhanced in isolated tracheal smooth-muscle membranes from hypersensitive guinea pigs. These results suggest that hypersensitive reactions are associated with the improved coupling of the stimulatory G protein (Gs) with adenylyl cyclase.

Topics: Adenylate Cyclase Toxin; Adenylyl Cyclases; Amino Acid Sequence; Animals; Cells, Cultured; Cholera Toxin; Enzyme Activation; GTP-Binding Proteins; Guanine Nucleotides; Guanylyl Imidodiphosphate; Guinea Pigs; Hypersensitivity; Isoproterenol; Lung; Membranes; Molecular Sequence Data; Ovalbumin; Pertussis Toxin; Protein Kinase C; Tetradecanoylphorbol Acetate; Trachea; Virulence Factors, Bordetella

In vivo uptake of the probe 51Cr-labeled EDTA from the jejunum of egg albumin (EA)-sensitized rats was compared with controls at baseline and after intraluminal antigen challenge. Probe recovery in blood was 60-80% greater in sensitized animals during the baseline period, suggesting that sensitization resulted in increased intestinal permeability. Sensitized, but not control, rats demonstrated a 15-fold increase in 51Cr-EDTA uptake after intraluminal antigen; no change occurred with an unrelated protein. Macromolecular recovery was also enhanced in sensitized animals, since serum levels of immunoreactive EA were elevated 14-fold compared with controls. Antigen challenge was accompanied by biochemical (protease release) and morphological (reduced numbers) evidence of mast cell degranulation in sensitized rats. The neurotoxin tetrodotoxin (applied directly to ligated jejunal segments) inhibited EA-induced uptake of 51Cr-EDTA and antigen. In isolated jejunum from sensitized rats, tetrodotoxin reduced secretory responses to luminal, but not serosal, antigen. These results indicate that neural factors may influence the uptake of molecules from the gut lumen during intestinal anaphylaxis.

Topics: Anaphylaxis; Animals; Cell Membrane Permeability; Electric Conductivity; Electrophysiology; Epithelium; Hypersensitivity; Immunoglobulin E; Jejunum; Male; Membrane Potentials; Muscle, Smooth; Ovalbumin; Rats; Rats, Sprague-Dawley; Tetrodotoxin

Peptide leukotrienes have been suggested to play an important role in bronchial asthma. As antigen-induced bronchoconstrictions, airway hyperreactivity, and pulmonary eosinophil accumulation are characteristics of the pathology of asthma, we investigated the effect of a peptide leukotriene receptor antagonist, ONO-1078, on these responses using guinea-pig models of asthma. Oral administration of ONO-1078 (3 mg/kg) significantly inhibited slow-reacting substance of anaphylaxis-mediated bronchoconstriction induced by i.v. administered ovalbumin. ONO-1078 (30-100 mg/kg), when administered orally both 1 h before and 4 h after ovalbumin challenge, significantly reduced immediate- and late-phase asthmatic responses, with peak responses occurring immediately and 5-11 h after challenge with inhaled ovalbumin. Oral administration of ONO-1078 significantly reduced the airway hyperreactivity (10-30 mg/kg) and the pulmonary eosinophil accumulation (30-100 mg/kg) observed 4 and 24 h after ovalbumin challenge, respectively. These results suggest that ONO-1078 may be of therapeutic use for bronchial asthma.

Topics: Acetylcholine; Acute-Phase Reaction; Animals; Asthma; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Bronchodilator Agents; Chromones; Disease Models, Animal; Guinea Pigs; Hypersensitivity; Immunization; Male; Ovalbumin; Phthalazines; Pulmonary Eosinophilia; Pyrilamine; SRS-A

The effect of heparin and poly-L-glutamate on the function of inhibitory M2 muscarinic autoreceptors on parasympathetic nerves in the lung was tested in antigen-challenged guinea pigs. After antigen challenge, M2 receptor function is decreased, thus increasing release of acetylcholine from the vagus and potentiating vagally induced bronchoconstriction. Guinea pigs were anesthetized, tracheostomized, vagotomized, paralyzed, and ventilated. Electrical stimulation of the vagi caused bronchoconstriction and bradycardia. In controls, pilocarpine attenuated vagally induced bronchoconstriction by stimulating neuronal M2 muscarinic receptors. Conversely, blocking these autoreceptors with gallamine potentiated vagally induced bronchoconstriction. In challenged animals the effects of both drugs were markedly reduced, confirming M2 receptor dysfunction. 20 min after heparin or poly-L-glutamate, the effects of both pilocarpine and gallamine on vagally induced bronchoconstriction were restored, demonstrating recovery of M2 receptor function. Neither heparin nor poly-L-glutamate affected vagally induced responses in control animals. Thus antigen-induced dysfunction of M2 receptors can be reversed by polyanionic polysaccharides (heparin) or polyanionic peptides (poly-L-glutamate). This suggests that a polycationic substance such as eosinophil major basic protein, cationic protein, or peroxidase may be responsible for antigen-induced pulmonary M2 receptor dysfunction.

Topics: Animals; Bronchial Provocation Tests; Bronchoconstriction; Gallamine Triethiodide; Guinea Pigs; Heparin; Hypersensitivity; Lung; Ovalbumin; Pilocarpine; Polyglutamic Acid; Receptors, Muscarinic; Vagus Nerve

Anti-inflammatory properties of zinc protoporphyrin disodium (Zn-PP-2Na) were studied. Zn-PP-2Na exhibits anti-allergic action against type III and IV reactions (passive Arthus reaction in rats and tuberculin-induced footpad reaction in mice), but does not affect type I and II reactions (homologous passive cutaneous anaphylaxis in mice and reversed cutaneous anaphylaxis in rats). Zn-PP-2Na also clearly inhibits type II collagen-induced arthritis in mice. The agent inhibits general arthritis symptoms, anti-type-II collagen antibody production and type II collagen-induced delayed type hypersensitivity (DTH) in arthritic mice. Zn-PP-2Na, however, did not affect carrageenin-induced paw edema and histamine- and serotonin-induced skin reactions in rats. Zn-PP-2Na inhibits IL-1-induced mouse lymphocyte proliferation, but does not affect PMA-induced O2- generation from guinea-pig neutrophil. These results indicate that Zn-PP-2Na inhibits type II collagen-induced arthritis in mice due to the antagonism of IL-1 activity and the inhibition of DTH against type II collagen.

Topics: Anaphylaxis; Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis; Arthus Reaction; Ascaris; Collagen; Edema; Hypersensitivity; Interleukin-1; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred DBA; Ovalbumin; Oxygen Consumption; Passive Cutaneous Anaphylaxis; Protoporphyrins; Rats; Rats, Wistar; Skin Tests

We examined whether eucapnic hyperventilation with dry air produces the bronchoconstriction in anesthetized, non-sensitized rabbits and in ovalbumin sensitized rabbits. Eucapnic hyperventilation challenge with dry air containing 5% CO2 at room temperature was performed with 4 non-sensitized and 7 sensitized rabbits by mechanical ventilation for 15 min (120 breaths/min, 7 ml tidal volume/kg body weight). Total lung resistance (RL) and dynamic compliance (Cdyn) were measured before and 0, 5, 15, and 30 min after hyperventilation. In non-sensitized rabbits, RL and Cdyn did not change significantly. However, in sensitized rabbits, RL increased maximally by 48.9% +/- 9.0% at 5 min, and then decreased to the baseline level at 30 min after challenge. Cdyn decreased maximally by 12.5% +/- 3.5% at 15 min after challenge. These changes were significantly different from the baselines (p < 0.05). Furthermore, to investigate the role of histamine on hyperventilation-induced bronchoconstriction (HIB) in sensitized rabbits, we performed the hyperventilation challenges in 5 sensitized rabbits with the pretreatment of H1-receptor antagonist (chlorpheniramine, 1 mg/kg, i.v.) and found that the maximum increment of RL was suppressed to 24.2% +/- 7.4% of the control, which was significantly lower than the maximal RL in nontreated sensitized rabbits (p < 0.05). We concluded that HIB occurs only in sensitized rabbits and that histamine may play an important role in the development of HIB in sensitized rabbits.

Topics: Airway Resistance; Animals; Bronchoconstriction; Carbon Dioxide; Chlorpheniramine; Histamine; Hypersensitivity; Hyperventilation; Lung Compliance; Male; Ovalbumin; Physical Exertion; Rabbits

In order to establish a safer and simpler antigen administration method in immunotherapy, we prepared biodegradable microspheres containing antigen and evaluated its safety and efficacy using guinea pigs. Poly (lactic/glycolic acid); (LGA) microspheres containing ovalbumin (OA) were fabricated by solvent evaporation. Over 70% of the OA was released from the microspheres within 3 days, and release was completed within 14 days in vivo. The local tissue reactions to the OA-LGA microspheres were apparently weaker than those to OA-alum. Repeated injections of high dose OA-LGA microspheres to OA-sensitized guinea pigs (high-LGA group) for 8 weeks at intervals of 2 weeks elicited an excellent therapeutic effect, i.e. a significant increase in the threshold value of antigen inhalation test, with a significant increase in IgG2 blocking antibody. The therapeutic efficacy of the high-LGA group was comparable to the conventional immunotherapy model (conventional group) and was superior to the antigen-alum model (alum group). We concluded that administration of antigen-LGA microspheres could become a new immunotherapeutic method for allergic disorders, being safer and requiring a lower frequency of antigen injections than the conventional method.

Topics: Animals; Antigens; Drug Delivery Systems; Guinea Pigs; Humans; Hypersensitivity; Immunoglobulin G; Immunotherapy; Injections; Lactates; Lactic Acid; Microspheres; Ovalbumin; Polyesters; Polyglycolic Acid; Polymers

The lungs of sensitized horses were exposed to aerosolized ovalbumin. Some horses (n = 4) were given ovalbumin in 1 lung only, whereas in others (n = 7), ovalbumin or vehicle were inoculated in the cranial, ventral, and caudal regions of the caudal lung lobe. Horses were exercised 5 hours after ovalbumin exposure. Immediately before exercise, endoscopy failed to reveal any abnormality. After exercise, endoscopic examination of horses subjected to unilateral ovalbumin exposure revealed extensive blood in airways leading to the exposed lung in all horses. Blood was not observed in the airways leading to the control lung. Mean (+/- SEM) minimum volume of the exposed and control lungs was 9.5 +/- 1.5 and 5.5 +/- 1.6 L, respectively; this difference was statistically significant (P less than 0.05). Bronchoscopy of horses subjected to regional ovalbumin or vehicle exposure and exercise revealed a small amount of blood-tinged fluid in the bronchi serving the regions of the lung inoculated with ovalbumin. Minimum volumes of such regions were not significantly different from one another. However, their minimum volume was significantly (P less than 0.05) larger than that of vehicle-inoculated regions. Gross and histologic examination confirmed inflammation and hemorrhage in the ovalbumin-exposed, but not the control lungs or lung regions. Thus, exercise can cause blood from an injured region of lung to appear in the larger airways. Regional differences in lung structure and function do not influence the appearance of blood in the airways.

Topics: Aerosols; Animals; Carbon; Coloring Agents; Exercise Test; Hemorrhage; Horse Diseases; Horses; Hypersensitivity; Lung; Lung Diseases; Ovalbumin; Physical Exertion; Staining and Labeling

1. SDZ PCO 400 evoked dose-related relaxation of isolated airway smooth muscle. For human bronchus precontracted by endogenous tone or addition of carbachol (10(-5) M), IC50 values were 1.74 microM and 1.82 microM respectively. With guinea-pig trachea contracted by endogenous tone, a comparable IC50 (1.79 microM) was observed, but no IC50 (less than 100 microM) could be determined following contraction by carbachol (10(-6) M). 2. Airway obstruction induced by intravenous bombesin in the anaesthetized ventilated guinea-pig was diminished by intravenous injection of SDZ PCO 400 (ID50 54 micrograms kg-1) or by introduction into the duodenum (ID50 1.0 mg kg-1). Inhalation of nebulized SDZ PCO 400 (0.1 mg kg-1) diminished airway obstruction due to intravenous injection of histamine (3.2-5.6 micrograms kg-1) for up to 20 min. 3. Increased bronchoconstrictor responses to bombesin (180-240 ng kg-1) following intravenous infusion of platelet activating factor (PAF) or (+/-)-isoprenaline, or to histamine (1.0-3.2 micrograms kg-1) following intravenous injections of immune complexes, were suppressed following concomitant intravenous infusion of SDZ PCO 400 (ID50 0.3 mg kg-1 h-1, 1.0 mg kg-1 h-1 and 0.1 mg kg-1 h-1 respectively). 4. Intravenous injection of SDZ PCO 400 (0.1 mg kg-1) effected transient (less than 10 min) inhibition of histamine-induced bronchospasm, yet diminished, for prolonged periods [up to 40 min] the enhanced bronchoconstrictor responses to histamine that followed intravenous injections of immune complexes.The capacity of SDZ PCO 400 to resolve such established airway hyperreactivity was prevented by prior intraduodenal instillation of a potassium channel antagonist, glibenclamide (30 mg kg-').5. In sensitized guinea-pigs, SDZ PCO 400 inhaled as a dry powder (5.7 mg kg-') suppressed development of allergic airway hyperreactivity to histamine (1.8-3.2;pg kg-', i.v.), but failed to diminish accumulation of eosinophils or other inflammatory cells within the airway lumen 24 h after inhalation of ovalbumin.6. Preincubation (30 min) of isolated sensitized trachea of guinea-pig with SDZ PCO 400 (10-5-10-4M) did not influence contractile responses to ovalbumin. However in anaesthetized sensitized guinea-pigs,insufflation of SDZ PCO 400 (1.25 mg) as a powder substantially diminished airway obstruction that followed inhalation of ovalbumin. This effect was prevented by prior vagal section.7. It is concluded that SDZ PCO 400 reduces airway obstruction not o

Topics: Airway Obstruction; Animals; Benzopyrans; Bronchial Hyperreactivity; Bronchodilator Agents; Cyclopentanes; Eosinophilia; gamma-Globulins; Guinea Pigs; Humans; Hypersensitivity; In Vitro Techniques; Lung; Muscle Relaxation; Muscle, Smooth; Ovalbumin; Parasympatholytics; Potassium Channels

A series of 6-alkyl- or 6-(cycloalkylalkyl)-[1,3,4]thiadiazolo[3,2- a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-ones 1b--o was synthesized from the corresponding 1,3,4-thiadiazol-5-amines 3b--o and the antiallergic activities of the products were evaluated. Among the compounds 6-(2-cyclohexylethyl)- [1,3,4]thiadiazolo[3,2-a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-one 1h, whose X-ray crystallographic stereostructure is shown, was found to be a promising new antiallergic agent, which has low toxicity and dual activity as a leukotriene D4 receptor antagonist and as an orally active mast cell stabilizer.

Topics: Animals; Female; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Lung; Muscle Contraction; Ovalbumin; Passive Cutaneous Anaphylaxis; Pyrimidines; Rats; Rats, Sprague-Dawley; SRS-A; Thiadiazoles; Triazoles; X-Ray Diffraction

The purpose of the study was to investigate the relationships between upper airways responses and pulmonary responses of two strains of highly inbred rats to inhaled antigen. To do this we measured the upper and lower airways resistance for 60 min after challenge of Brown-Norway rats (BN; n = 13) and an inbred rat strain (MF; n = 11), derived from Sprague-Dawley, with aerosolized ovalbumin (OA). Rats were actively sensitized with OA (1 mg sc) using Bordetella pertussis as an adjuvant. Two weeks later the animals were anesthetized and challenged. Tracheal pressure, esophageal pressure, and airflow were measured, from which total pulmonary resistance was partitioned into upper airway and lower pulmonary resistance (RL). The peak upper airway response to inhaled OA was similar in BN (1.89 +/- 0.66 cmH2O.ml-1.s; n = 7) and MF (2.85 +/- 0.68 cmH2O.ml-1.s; n = 6). The lower airway response to OA challenge was substantially greater in BN, and RL changed from 0.07 +/- 0.01 to 0.34 +/- 0.13 (n = 6; P < 0.05). The MF did not have any significant increase in RL after challenge; the baseline RL was 0.12 +/- 0.02 and only reached a peak value of 0.15 +/- 0.05 (n = 5; P = NS). Lower airway responsiveness of BN (n = 10) to serotonin, an important mediator early allergic airway responses, was similar to MF (n = 7).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Administration, Inhalation; Aerosols; Airway Resistance; Animals; Antigens; Hypersensitivity; Immunoglobulin E; Male; Mast Cells; Methacholine Compounds; Ovalbumin; Rats; Rats, Sprague-Dawley; Respiratory Physiological Phenomena; Respiratory System; Serotonin

Peripheral blood lymphocytes from nonallergic individuals acquired responsiveness to interleukin 2 (IL2) after stimulation with ovalbumin (OVA) or Dermatophagoides farinae (Df) antigens when they were pretreated with the CD45RA antibody, which has been shown to define the suppressor inducer subset of CD4+ cells and also to block its suppressor activity. The effect provided by the CD45RA antibody was lost if the lymphocytes had initially been activated with the OVA of Df antigens. The magnitude of the responses was comparable to the allergen-induced responses observed in OVA- or Df-sensitized lymphocytes from allergic patients. The pre-existing IL2 responsiveness in the patients was not increased by the CD45RA antibody pretreatment. However, the CD45RA antibody pretreatment gave rise to Df-induced IL2 responsiveness in the lymphocytes of the patients sensitized with OVA but not with Df; conversely, OVA-induced IL2 responsiveness was enhanced in Df- but not in OVA-sensitized lymphocytes. The CD45RA antibody apparently acts on CD4+ T cells, but not on CD8+ T cells, to induce the IL2 response. A further dissection of normal CD4+ T cells indicated that CD4+45RA- T cells preferentially respond to IL2 after stimulation with OVA or Df antigens. Since normal CD4+45RA+ T cells did not show antigen-induced IL2 responsiveness even after pretreatment with the CD45RA antibody, it is unlikely that the CD45RA antibody stimulates CD4+45RA+ T cells to become responsive to IL2 after antigenic challenge. Alternatively, CD4+45RA+ T cells may modulate the activity of CD4+45RA- T cells, which are potentially responsive to IL2 by antigenic stimulation and thus provide tolerance in nonallergic lymphocytes. Collectively, a defective suppressor activity of CD4+45RA+ T cells may exist in patients with hen-egg allergy and/or bronchial asthma, which may cause lymphocytes to be hyperreactive to OVA or Df antigens.

Topics: Adult; Allergens; Antibodies; Antibodies, Monoclonal; Antigens, CD; Antigens, Dermatophagoides; CD4 Antigens; Histocompatibility Antigens; Humans; Hypersensitivity; Interleukin-2; Kinetics; Leukocyte Common Antigens; Lymphocyte Activation; Lymphocytes; Ovalbumin; Receptors, Interleukin-2; T-Lymphocyte Subsets; T-Lymphocytes

ACE-inhibitors have for some time been used in the treatment of hypertension. Apart from inhibiting the conversion of angiotensin I to II, the drugs also affect the metabolism of some inflammatory agents, like bradykinin and substance P. Egg albumin (EA)-sensitized guinea pigs were pretreated with the ACE-inhibitors. Measurement of flare and wheal areas induced by an intradermal injection of EA, showed that enalaprilat significantly increased, whereas cilazaprilat slightly decreased, the reaction area. Enalaprilat also showed an enhancement in histamine and substance P (SP) contents in the skin. In vitro incubation of guinea pig biopsies with enalaprilat potentiated EA- but not SP-induced histamine release. The EA-induced effect was abolished if the animals were pretreated with capsaicin. The conclusion is that cilazaprilat, in contrast to enalaprilat, does not potentiate inflammatory reactions in the guinea pig.

Topics: Angiotensin-Converting Enzyme Inhibitors; Animals; Antigens; Cilazapril; Dermatitis; Enalaprilat; Female; Guinea Pigs; Histamine Release; Hypersensitivity; Ovalbumin; Pyridazines

The response to antigen (trinitro-phenyl-haptenized ovalbumin) and the modulatory role of several antiallergic drugs was studied in isolated hearts from actively sensitized rats. Antigen induced a triphasic effect on coronary flow (CF) and left ventricular pressure (LVP) characterized by short-term increase (0-1.5 min = phase 1) and a severe decrease (1.5-7.5 min = phase 2) followed by a less pronounced long-lasting decrease (7.5- greater than 20 min = phase 3). The first phase was accompanied with a substantial release of 5-hydroxytryptamine (5-HT), histamine, and leukotrienes measured in cardiac effluents. The histamine2 (H2)-receptor antagonist cimetidine (60 microM) reversed the antigen-induced increase in CF to a decrease. In contrast, H1-receptor blockade by mepyramine (6 microM) had no effect. Methysergide (10 microM) and ketotifen (0.1 microM) evoked a mild suppression during all three phases. Indomethacin (10 microM) was almost inactive while tolfenamic acid (1 microM) was slightly active in this respect during phase 2. Addition of the 5-lipoxygenase inhibitor AA 861 (1 microM) resulted in complete suppression of the antigen-induced decrease in CF. The leukotriene antagonist FPL 55712 (5 and 50 nM) evoked a dose-dependent suppression with respect to the anaphylactic phases 2 and 3. A similar reduction was obtained with sodium cromoglycate (1 mM). AA 861, FPL 55712, and sodium cromoglycate also suppressed the antigen-induced decrease in LVP. The antigen-induced histamine release was not affected by the aforementioned drugs. Our results provide evidence that H2-receptor blockade during cardiac anaphylaxis enhances coronary constriction and may be detrimental in this condition. On the other hand, leukotriene antagonists and 5-lipoxygenase inhibitors may exert beneficial effects during cardiac anaphylaxis. Further experiments in this area are needed to clarify the precise role of mast cell-generated mediators in cardiac anaphylaxis possibly leading to new therapeutic approaches in this life-threatening disorder.

Topics: Anaphylaxis; Animals; Antigenic Modulation; Antigens; Chromatography, High Pressure Liquid; Coronary Circulation; Heart; Histamine; Histamine H1 Antagonists; Histamine H2 Antagonists; Histamine Release; Hypersensitivity; Immunoglobulin E; In Vitro Techniques; Leukotrienes; Lipoxygenase Inhibitors; Male; Myocardium; o-Phthalaldehyde; Ovalbumin; Radioimmunoassay; Rats; Rats, Inbred Strains; Serotonin; Spectrometry, Fluorescence

It has been reported previously that acute allergic reactions in the anaesthetized guinea-pig do not evoke changed airway reactivity to histamine, animals sensitized to ovalbumin in aluminum hydroxide and exposed to allergen intravenously or by inhalation develop increased responsivity to histamine that persists for several hours. Animals that have been sensitized passively, by intravenous injection of antibody and exposed to allergen by intravenous injection, or which receive freshly prepared immune complexes by intravenous injection develop comparable airway reactivity.

Topics: Animals; Antigen-Antibody Complex; Guinea Pigs; Histamine; Hypersensitivity; Immune Sera; Immunization; Male; Ovalbumin; Respiratory Hypersensitivity

Responses to bilateral vagal nerve stimulation, to field stimulation, and to exogenous methacholine and histamine were compared in tracheas isolated from (a) saline injected (i.p.) and saline-aerosol exposed guinea pigs (control), (b) ovalbumin-sensitized and saline-aerosol exposed (sensitized) guinea pigs, and (c) ovalbumin-sensitized and 2% ovalbumin-aerosol exposed (challenged) guinea pigs. Tracheal pressor responses (cmH2O; 1 cmH2O = 98.1 Pa) to nerve and field stimulation, and maximal responses to methacholine and histamine were significantly increased in animals from group c compared with groups a and b. Dose-response lines in response to the two agonists, expressed as percent maximal contraction, did not differ among the groups. The M1 antagonist pirenzepine (0.1-10 nM) selectively reduced responses to nerve stimulation in all three groups. The M2 antagonist gallamine potentiated responses to nerve or field stimulation in all three groups. We conclude that M1, M2, and M3 muscarinic receptor functioning is similar in control and ovalbumin-sensitized guinea pigs. Changes in post-receptor transduction mechanisms may mediate the increased responsiveness noted in animals from group c.

Topics: Aerosols; Animals; Asthma; Bethanidine; Electric Stimulation; Female; Gallamine Triethiodide; Guinea Pigs; Histamine; Hypersensitivity; In Vitro Techniques; Indomethacin; Methacholine Compounds; Ovalbumin; Pirenzepine; Propranolol; Receptors, Muscarinic; Trachea; Yohimbine

A rat model has been developed to examine the possible role of homocytotropic antibodies in initiating or exacerbating synovial inflammation. The technique, passive synovial anaphylaxis, involves passively sensitizing rat knee joints with specific IgE, then challenging intravenously with the corresponding antigen while monitoring for signs of inflammation. Swelling of the sensitized joints reached maximum 2 h after the challenge, then gradually decreased to prechallenge levels by 24 h. Radioisotopic joint scans detected a passive synovial anaphylaxis induced increase in local blood flow and exudation within the joints. The degree of swelling correlated directly with the amount of antigen specific IgE in the sensitizing serum, and individual joints remained sensitized for up to 36 days after the IgE injection.

Topics: Animals; Antibody Formation; Dinitrophenols; Enzyme-Linked Immunosorbent Assay; Hypersensitivity; Immunoglobulin E; Knee Joint; Ovalbumin; Rats; Rats, Inbred BN; Rats, Inbred Strains; Serum Albumin, Bovine; Synovitis; Time Factors; Whooping Cough

Bronchoconstriction (BC) is the main feature of anaphylaxis in the guinea pig. Since LPS induces lung inflammation and antigen-induced BC depends on the endogenous formation of histamine and arachidonate metabolites, we studied whether LPS might modulate antigen-induced BC. Guinea pigs were sensitized subcutaneously with 10 micrograms ovalbumin (OA) on days 0 and 14. LPS (100 micrograms/kg) was injected intravenously on day 21, and daily injections of LPS were continued before the antigenic challenge on day 22, 23, 24, or 25. Intratracheal injection of 100 micrograms OA induced an abrupt and reversible BC. Single or repetitive injections of LPS reduced BC. LPS is likely to reduce the OA-induced BC by affecting the histamine-dependent component of BC, since (a) LPS induced a partial degranulation of lung mast cells; (b) BC is reduced by mepyramine, an histamine receptor antagonist; (c) LPS did not affect BC in mepyramine-treated guinea pigs; (d) LPS reduced histamine release by OA-stimulated guinea pig lungs in vitro. Moreover, the in vitro OA-induced production of arachidonate metabolites was also reduced by LPS. The decreased formation of TXB2 was not only secondary to a reduced release of histamine, since LPS inhibited TXB2 formation in the presence of mepyramine. Finally, the FMLP-induced BC and mediator release were inhibited by LPS, whereas the platelet activating factor-induced pulmonary responses were not. Thus, the protective effect of LPS is not antigen-specific and does not result from a general desensitization. These studies indicate that a single dose of LPS reduces the antigen-induced BC by reducing histamine release from lung mast cells, although a decreased formation of eicosanoids may contribute to the protective effect of LPS.

Topics: Animals; Antigens; Aspirin; Bronchoconstriction; Cell Degranulation; Endotoxins; Escherichia coli; Guinea Pigs; Hemodynamics; Histamine; Hypersensitivity; Leukopenia; Lipopolysaccharides; Mast Cells; Microscopy, Electron; N-Formylmethionine Leucyl-Phenylalanine; Ovalbumin; Platelet Activating Factor; Pyrilamine; Thromboxane B2

Hyperreactivity and bronchial inflammation resulting from active anaphylactic shock induced by aerosol have been studied in guinea pigs after sensitization by intramuscular injection of large-dose ovalbumin or aerosol ovalbumin. When animals were sensitized by i.m. injection of 30 mg/kg ovalbumin, hyperreactivity to inhalation of histamine was obtained 1-3 h after shock. In bronchoalveolar lavage (BAL) fluid an increase in the number of eosinophils (6-48 h after shock) and neutrophils (6-24 h) was observed. When guinea pigs were sensitized by aerosol route, the hyperreactivity to histamine inhalation appeared 1-6 h after shock. In BAL fluid the number of mononuclear cells dropped (1-3 h) and then increased (24-48 h); the number of neutrophils (6-48 h) and eosinophils (24-48 h) increased. The results observed during these two types of sensitization were compared to those obtained after sensitization by injection of a large dose of ovalbumin mixed with Freund's complete adjuvant.

Topics: Aerosols; Anaphylaxis; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Eosinophils; Guinea Pigs; Hypersensitivity; Injections, Intramuscular; Male; Neutrophils; Ovalbumin

Interleukin 2 (IL2) responsiveness of ovalbumin (OVA)-stimulated lymphocytes from patients with hen-egg allergy was studied. The number of viable cells of 5-day cultured lymphocytes stimulated with OVA was increased by an additional three days incubation with recombinant IL2. This phenomenon was not observed when the lymphocytes of patients allergic to OVA but not to Dermatophagoides farinae extract antigen (Df) were stimulated with Df. Normal lymphocytes stimulated with OVA expressed Tac antigen (low affinity IL2 receptor) but, in contrast to those from the allergic patients, did not absorb nor respond to IL2. The induction of OVA-specific IL2 responsiveness in patient lymphocytes was markedly suppressed on the addition of culture supernatant from OVA-stimulated normal T cells, but Df-specific IL2 responsiveness of the lymphocytes from Df-sensitized patients with bronchial asthma was not suppressed by the same supernatant. The supernatant of lymphocytes from allergic patients did not show such suppressive effect. The patient lymphocytes whose IL2 responsiveness was suppressed with the supernatant from normal lymphocytes still expressed Tac antigen. These observations suggest that the culture supernatant of normal T lymphocytes stimulated with OVA contained an antigen-specific factor suppressing the induction of IL2 responsiveness of OVA-stimulated patient lymphocytes. The production of such a suppressive factor was impaired in the patient, and further, the factor may have inhibited the triggering signal of IL2 receptors having absorbed IL2. The existence of some allogeneic barrier between the factor(s) and patient lymphocytes was suggested, since the supernatant from OVA-stimulated normal T cells did not necessarily suppress the response of all patients tested.

Topics: Absorption; Adult; Cells, Cultured; Child; Child, Preschool; Eggs; Epitopes; Female; Humans; Hypersensitivity; Infant; Interleukin-2; Lymphocytes; Male; Ovalbumin; Receptors, Interleukin-2; Reference Values

The mechanisms underlying the different sensitivity to respiratory anaphylaxis were studied in two inbred strains of guinea pigs with high (IMM/S) or low (IMM/R) sensitivity to ovalbumin (OA) aerosol-induced respiratory anaphylaxis. Guinea pigs were immunized with OA in Freund's complete adjuvant and lymphocyte stimulation tests were done with cells from the tracheobroncheal lymph nodes (TBL) draining the airways, from peripheral blood lymphocytes and from the axillary and inguinal lymph nodes. The TBL of the IMM/R strain had a significantly lower response than those of the IMM/S strain. This selectively low response, which was not antigen specific, could be increased by cyclophosphamide pretreatment. The high TBL response in the IMM/S strain was decreased by OA inhalations to the level of the IMM/R strain. OA inhalations decreased the IgE response in the IMM/S but not the IMM/R strain. Different interpretations of these findings are discussed, with emphasis on the possibility that an inherent suppressor cell population in the TBL are involved.

Topics: Anaphylaxis; Animals; Asthma; Bronchi; Bronchial Provocation Tests; Cyclophosphamide; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Inbreeding; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Ovalbumin; Trachea

The pharmacodynamics of Amlexanox (AA-673), an azoxanthone derivative recently developed as an ocular anti-allergic drug, were studied in normal and anaphylactic rat conjunctiva. Ocular anaphylaxis was induced by topical application of dithiothreitol and antigen challenge (egg albumin). The drug concentration was measured at 5, 30, 60 and 120 min after instillation of AA-673 0.25% ophthalmic solution. Histamine concentrations in the pretreated and untreated anaphylactic conjunctiva were compared. In both groups the drug concentration decreased exponentially with time, showing a marked delay in the anaphylactic group. There was a significant difference in histamine values between the pretreated and untreated groups.

Topics: Aminopyridines; Animals; Chromatography, High Pressure Liquid; Conjunctiva; Dithiothreitol; Histamine; Hypersensitivity; Male; Ovalbumin; Rats; Rats, Inbred Strains

Peripheral blood mononuclear cells of patients allergic to honey bee venom were stimulated with denatured bee venom phospholipase A2, and the antigen-activated T cells were propagated for 4 days by human IL-2 in the presence or absence of recombinant human lipocortin I. Upon antigenic stimulation with the denatured phospholipase A2 and autologous monocytes or by cross-linking of CD3 by anti-CD3 antibody, the activated T cells, which had propagated by IL-2 alone, formed N-glycosylated IgE-binding factors and glycosylation enhancing factor (GEF), while those propagated in the presence of lipocortin formed unglycosylated IgE-binding factors and glycosylation inhibiting factor (GIF). The GEF and GIF formed by the antigen- or anti-CD3-stimulated T cells had affinity for bee venom phospholipase A2 and could be purified by using anti-lipomodulin Sepharose. In the mouse lymphocyte system, the major cell source of GIF is antigen-specific suppressor T cells, and the antigen-binding GIF from the cells suppressed the in vivo antibody response in an antigen (carrier)-specific manner. In view of the findings in the mouse system, the present results may provide an immunological maneuver to generate allergen-specific suppressor T cells, and to obtain allergen-specific suppressor factor from T cell populations in the peripheral blood of allergic patients.

Topics: Annexins; Bee Venoms; Calcium-Binding Proteins; Humans; Hypersensitivity; Interleukin-2; Lymphocyte Activation; Lymphokines; Ovalbumin; Phospholipases; Phospholipases A; Phospholipases A2; Prostatic Secretory Proteins; Suppressor Factors, Immunologic; T-Lymphocytes, Regulatory

The role of platelet-activating factor (PAF) in Ag-induced airway hyperresponsiveness was evaluated in a guinea pig model using the PAF antagonist SDZ 64-412. Repeated OVA challenge by aerosol (twice weekly x 4 wk) of previously sensitized guinea pigs produced striking airway hyperresponsiveness as determined by pulmonary resistance changes to increasing doses of inhaled acetylcholine given 3 days after the last OVA challenge. Each OVA challenge produced significant hypoxia that was unaffected by oral pretreatment with 20 mg/kg SDZ 64-412, 2 h before each challenge (pO2 = 35 +/- 2 mm Hg for OVA alone vs 40 +/- 6 mm Hg for SDZ and OVA groups, respectively). SDZ 64-412 pretreatment abolished the airway hyperresponsiveness resulting from repeated Ag challenge. Morphometric analysis revealed that SDZ 64-412 treatment had no effect on the increased numbers of eosinophils that infiltrated the airways of OVA-challenged guinea pigs. These results suggest that PAF may be a primary mediator of airway hyperresponsiveness, but not acute bronchoconstriction, induced by repeated Ag challenge. This activity of PAF appears independent of eosinophil recruitment to airways.

Topics: Acetylcholine; Airway Resistance; Animals; Bronchial Provocation Tests; Bronchial Spasm; Connective Tissue Cells; Eosinophils; Epithelial Cells; Guinea Pigs; Hypersensitivity; Isoquinolines; Ovalbumin; Platelet Activating Factor; Platelet Aggregation; Respiratory System

Although mast cells are sometimes considered to play a minor role in hypersensitivity reactions, we used a histological method to show the modifications in the guinea pig lung mast cell population in the course of such reactions. We sensitized guinea pigs with ovalbumin and studied the effect of the challenge with and without corticosteroid treatment. We observed that the mast cell count was not modified after sensitization but was decreased after challenge. Twenty-four hours after challenge, the number of mast cells returned to the control value, indicating a renewal of the mast cell pool. A second challenge, 1 week after the first, did not provoke the same mast cell degranulation, suggesting a non-responsiveness to aerosol antigen. Betamethasone dipropionate treatment protected mast cells against challenge: in treated guinea pigs, mast cell degranulation was prevented, and we did not observe any change in mast cell count after challenge. The present study was useful to show an effect of corticosteroids on mast cell degranulation in immediate hypersensitivity reactions in vivo.

Topics: Adrenal Cortex Hormones; Animals; Betamethasone; Bronchi; Cell Count; Guinea Pigs; Hypersensitivity; Male; Mast Cells; Ovalbumin

This study examined the electrophysiological responses to antigen and to various stimuli in jejunal mucosa from rats sensitized to egg albumin with alum and pertussis adjuvants. Luminal antigen caused an immediate increase in short-circuit current, a measure of net ion transport, which was one of three different patterns. All were inhibited by the chloride channel blocker diphenyl-2-carboxylate, by chloride-free buffer, and by doxantrazole, a mast cell stabilizer. Depending on the pattern, the histamine-1 antagonist diphenhydramine, the 5-hydroxytryptamine-2 antagonist ketanserin, and the cyclooxygenase inhibitor piroxicam also reduced the responses. A neural component was indicated by inhibition of the responses to luminal antigen by the neurotoxin tetrodotoxin and by neonatal capsaicin treatment, which depletes substance P-containing nerves. In the absence of antigen, histamine and substance P caused increases in short-circuit current; the magnitude of these changes was significantly greater in tissues from sensitized animals than in controls. These data suggest that sensitization itself may result in hypersecretory responses to some inflammatory mediator and neurotransmitter substances.

Topics: Animals; Antigens; Biological Transport; Capsaicin; Diphenhydramine; Electrolytes; Hypersensitivity; Intestinal Mucosa; Ions; Jejunum; Ketanserin; Ovalbumin; Piroxicam; Rats; Rats, Inbred Strains

We have previously demonstrated that Cyclosporin A (CyA), a T lymphocyte-selective immunosuppressive agent, reduced the delayed-phase bronchial eosinophil infiltration after antigen challenge in a guinea pig model of asthma. In the present study, we studied the effects of CyA on antigen-induced late asthmatic response (LAR) and bronchial hyperresponsiveness following LAR. Guinea pigs immunized by repeated exposure to aerosolized ovalbumin (OA) were intravenously given metopirone, a cortisol synthesis inhibitor, 24 hours before and 30 minutes before antigen challenge, and to prevent death from immediate severe bronchoconstriction, chlorpheniramine maleate was also injected. After antigen challenge with high dose of OA, LAR occurred in twelve of fifteen animals (80%) and the bronchial responsiveness to acetylcholine was significantly increased. However, when guinea pigs were treated with CyA from the beginning of immunization period, the development of LAR was completely inhibited, although similar magnitude of immediate bronchoconstriction was observed, and a subsequent increase in bronchial responsiveness was partially but significantly blocked. Since CyA has been shown to suppress activation of guinea pig T lymphocytes and their production of lymphokines, these results suggest that T cell factor(s) may be important for the elicitation of LAR and the antigen-induced bronchial hyperresponsiveness.

Topics: Animals; Asthma; Bronchi; Cyclosporins; Guinea Pigs; Hypersensitivity; Male; Ovalbumin

Arachidonic acid (AA) and ovalbumin (OA) were used to induce contractions of sensitized guinea pig tracheal spiral (indomethacin-pretreated) and lung parenchymal strip preparations. This model was used to examine the properties of three leukotriene (LT) D4 antagonists and a platelet-activating factor (PAF)-acether receptor antagonist. The three LTD4 antagonists, L-649,923, FPL 57231, and LY163443, inhibited AA-induced contractions of indomethacin-pretreated tracheal spirals selectively. The PAF-acether antagonist, L-652,731, did not inhibit AA-induced contractions of either trachea or parenchyma. This confirmed that AA-induced contractions of trachea involved release and activity of LTD4. The LTD4 antagonists and L-652,731 partially inhibited OA-induced contractions of both trachea and parenchyma. When L-649,923 and L-652,731 or FPL 57231 and L-652,731 were combined, an additive inhibitory effect on OA-induced contractions was observed. When LY163443 and L-652,731 were combined, the inhibitory effect was synergistic. This may be due to the additional effect of LY163443 to inhibit phosphodiesterase. Total inhibition of OA-induced contractions was obtainable with relatively low concentrations when a LTD4 and PAF-acether antagonist were combined. These results suggested that LTD4 and PAF-acether may be the two major mediators in our model of allergic bronchospasm. The LTD4 and PAF-acether antagonists had the capacity to decrease baseline tone, even on tissues that were already relaxed with indomethacin, suggesting that LTD4 and PAF-acether may contribute to intrinsic tone in airway smooth muscle.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Indomethacin; Lung; Muscle Contraction; Muscle Relaxation; Muscle, Smooth; Ovalbumin; Platelet Activating Factor; Respiratory System; SRS-A; Trachea

The occurrence of beta-adrenoceptor desensitisation in clinical experience, after long term therapy with specific beta 2-agonists, is still an open question, even if this phenomenon is easily observed in different experimental models. Since the majority of these experiments have been performed in normal animals, we investigated the possible occurrence of beta-adrenoceptor desensitisation in the ovalbumin actively sensitised guinea-pig model of experimental asthma. The isoproterenol concentration-response curves performed in pilocarpine contracted guinea-pig trachea in vitro were shifted to the right by the beta-adrenoceptor desensitisation procedure, which was achieved by the in vitro administration of isoproterenol (10(-5) M x 2 times x 20 min each) both in normal and ovalbumin sensitised tissues. The same desensitisation procedure markedly affected epinephrine-relaxing capacity in both normal and ovalbumin actively sensitised guinea-pig tracheae. However, the ovalbumin sensitised tissue seemed to be more sensitive than normal to the specific beta 2-agonist procaterol; in parallel the beta 2-mediated relaxation was more impaired by isoproterenol-induced beta-adrenoceptor down regulation in ovalbumin sensitised trachea when compared to normal. Similar results have been obtained using salbutamol as the beta 2-adrenoceptor desensitising agent. The changes in beta-adrenoceptor reactivity between normal and ovalbumin sensitised guinea-pig tracheae seemed to depend on the active sensitisation process. No difference in the degree of beta-adrenoceptor down regulation was observed in passively ovalbumin sensitised guinea-pig trachea as compared to normal. These data suggest that, in this model of experimental asthma, beta-adrenoceptor reactivity is in some way modified and that this phenomenon might contribute to the genesis of asthma.

Topics: Animals; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Male; Ovalbumin; Receptors, Adrenergic, beta; Trachea

Breast milk samples were collected from 152 women during the first week after delivery. The levels of IgG and IgA antibodies to beta-lactoglobulin, ovalbumin and gliadin were assessed with an enzyme-linked immunosorbent assay (ELISA). The breast milk antibody levels did not differ significantly between mothers on a strictly cow's milk and egg-free diet, and mothers taking these foods. Moreover, the colostral food antibody levels did not differ significantly between atopic and non-atopic mothers. Neither was there any correlation between the colostral antibody levels and the development of atopic disease in the baby. I conclude that maternal antigen avoidance during late pregnancy does not affect the food antibody levels in colostrum. High levels of food antibodies in a colostrum sample seem not to offer protection against food allergy in the child.

Topics: Animals; Colostrum; Diet; Food Hypersensitivity; Gliadin; Humans; Hypersensitivity; Infant; Infant, Newborn; Lactoglobulins; Milk; Milk, Human; Ovalbumin; Prospective Studies

Local bronchial mucosal hypersensitivity following antigen feeding was studied in the guinea pig. Groups of 6 animals were fed 1% ovalbumin (OA) in tap water or tap water without antigen (control group) for different feeding periods (14, 28, 42, and 56 days). Inhalative provocations with increasing concentrations of OA (0.5-8% OA) were performed at the end of each feeding period followed by body plethysmographic measurement of airway obstruction. Specific bronchial hypersensitivity to inhaled OA was not found in the control group, whereas specific bronchial reactivity to OA, described as reactivity index, was significantly different from the control group after 14 (p less than 0.05), 28 (p less than 0.001) and 42 days (p less than 0.01) of feeding. No difference to the control group was found after 56 days of feeding. Anti-OA IgG total and IgG1 in serum and bronchoalveolar lavage fluid were increased in OA-fed animals reaching maximal concentrations at day 28 of the feeding period. We conclude that oral feeding of a 1% solution of OA can induce a transient state of local hypersensitivity to inhaled antigen in the guinea pig as manifested by bronchoconstriction on OA inhalation and increased concentrations of local and systemic specific antibodies. This period of local hypersensitivity is followed by tolerance.

Topics: Aerosols; Animals; Bronchi; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Female; Guinea Pigs; Hypersensitivity; Immunoglobulin G; Ovalbumin; Plethysmography; Time Factors

The effect of glucocorticosteroid (GCS) treatment on ovalbumine-induced IgE-mediated immediate and late allergic response was studied in sensitized guinea pigs. The results show that the GCS budesonide (BUD) inhibits the allergen-induced IgE-mediated immediate and late bronchial obstruction. The effect on the early reaction is correlated to the inhibition of leukotrienes and histamine release. The importance of mediator release inhibition for the antianaphylactic effect of GCS is discussed. In examining the effect on the late reaction, it was found that BUD had to be present during the early reaction but did not inhibit the early reaction. Furthermore, the effect on the late reaction was correlated to the inhibition of vascular leakage but not to the infiltration of inflammatory cells as examined in bronchoalveolar lavage. The results indicate that some triggering factors important for the development of the late reaction are released during the early reaction. Inhibition of the release of that factor or the activation of inflammatory cells by that factor might be the mechanism behind the antiinflammatory activities of GCS.

Topics: Anaphylaxis; Animals; Antigens; Bronchi; Bronchial Provocation Tests; Budesonide; Cromolyn Sodium; Dimercaprol; Guinea Pigs; Histamine Release; Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulin E; Leukotriene B4; Lung; Ovalbumin; Peptide Hydrolases; Pregnenediones; SRS-A

The effect of sulfur dioxide (SO2) exposure on local bronchial sensitization to inhaled antigen was studied in the guinea pig. Exposure to SO2 (0.1 to 16.6 ppm) was performed in a 20 L exposure chamber for 8 hours on 5 consecutive days, while temperature, moisture, and concentration of SO2 were monitored and kept constant. SO2 concentrations were measured hourly by Schiff's reaction. On the last 3 days, SO2 exposure was followed by inhalation of nebulized ovalbumin (OA) for 45 minutes. One week later, specific bronchial provocation with inhaled OA (0.1%) followed by plethysmographic measurements of airway obstruction were performed every 2 days during a 2-week period. Specific antibodies against OA were measured in serum and bronchoalveolar fluid by a direct enzyme immunoassay. The SO2-exposed group (N = 17) demonstrated 67% to 100% positive bronchial reactions to inhaled OA, depending on the concentration of SO2, whereas the control group without previous SO2 exposure (N = 14) demonstrated bronchial reactions in only one animal (7%: p less than 0.05). The degree of bronchial obstruction was significantly higher in the exposed group, compared to the control group, for all SO2 concentrations (p less than 0.05). OA-specific antibodies in serum and bronchoalveolar fluid increased in SO2-exposed groups significantly, compared to the control group (p less than 0.05). It is concluded from these results that exposure to SO2 in low and medium concentrations can facilitate local allergic sensitization in the guinea pig.

Topics: Aerosols; Air Pollutants; Animals; Antibodies; Bronchi; Bronchial Provocation Tests; Dose-Response Relationship, Drug; Guinea Pigs; Hypersensitivity; Immunoglobulin G; Ovalbumin; Respiratory Function Tests; Sulfur Dioxide

554 L, a synthetic histamine depressant, was first made in September 1956. In spite of the age of this compound, it seemed to us that it may be of interest to examine it for possible experimental general and localised bronchial antianaphylactic activity. In the first experiments, the results showed an anti-anaphylactic effect in guinea pigs that had been sensitised to cow milk. The second study proved that there was an anti-anaphylactic effect locally on the bronchii by an aerosol of 554 L or Hypostamine on guinea pigs that were sensitive to OVA. These results may produce development of this old compound in delayed release and aerosol presentations.

Topics: Animals; Bronchial Provocation Tests; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Isoquinolines; Milk; Ovalbumin

Inhaled ozone was found to exert an enhancing effect for allergic lung sensitization when mice contracted an aerosolized allergen. The animals were exposed to ozone concentrations of 0.24, 0.16, 0.13, and 0.10 ppm. After 4 days of continuous ozone exposure, the mice had allergen contact from an aerosolized solution of ovalbumin. The animals were then maintained in ambient air for several days before the cycle of ozone and aerosolized allergen was repeated over four allergen contact cycles. Mice were rested in ambient air for a week after the last allergen contact, and they were then tested for allergic sensitization by the intravenous injection of 2 mg of ovalbumin to induce anaphylactic shock in allergic individuals. The control groups of mice were maintained in ambient air throughout the experiment, but they experienced identical allergen contact with the ozone-exposed mice. The phenomenon of allergic enhancement from ozone inhalation was detected at 0.24, 0.16, and 0.13 ppm of ozone. The enhancing effect disappeared at 0.10 ppm of ozone. The study indicated a potential for increasing the number of allergically sensitized individuals when various allergens are inhaled during periods of high ozone exposure with the consequent adverse changes on respiratory membranes. The significance to human health of the allergic enhancement phenomenon by ozone needs investigation.

Topics: Aerosols; Allergens; Anaphylaxis; Animals; Antigens; Female; Hypersensitivity; Lung Diseases; Ovalbumin; Ozone; Rats

Respiratory histaminergic and cholinergic receptor function was investigated in isolated tracheal spirals of guinea pigs receiving different diets. Comparison was made between saline treated (controls) and Haemophilus influenzae treated animals in non sensitized conditions, the latter being a model for bronchial hyperreactivity, and in sensitized conditions, being a model for allergen induced bronchial hypersensitivity. The different semi-synthetic diets (35 energy% fat), varying in linoleic acid content (5.85, 11.25 and 22.05 en% fat) and one diet with low linoleic acid (3.55 en%) in which linolenic acid was added additionally (5.30 en%), exerted profound effects on tracheal reactivity to histamine. In sensitized animals the maximal induced histamine contraction was significantly diminished in the dietary group receiving 5.85 en% linoleic acid as compared with the other dietary groups (35% decrease in the H. influenzae-treated, 20-30% decrease in saline treated animals). Results in non-sensitized animals were similar, though less pronounced. No effect on food intake or growth of the animals could be demonstrated during the six week experimental periods. The carbachol induced contraction of the tracheal spirals of sensitized animals receiving low linoleic acid was also significantly decreased as compared to the other dietary groups (30% for saline treated and 20-30% for H. influenzae-treated animals). No difference in carbachol responsiveness could be detected between the different dietary groups under non-sensitized conditions. The results are discussed in view of the current concepts for bronchial hyperreactivity, especially in relation to eicosanoid involvement.

Topics: alpha-Linolenic Acid; Animals; Bronchial Diseases; Carbachol; Dietary Fats; Fatty Acids, Unsaturated; Guinea Pigs; Haemophilus influenzae; Histamine; Hypersensitivity; Linoleic Acid; Linoleic Acids; Linolenic Acids; Male; Muscle Contraction; Ovalbumin; Receptors, Cholinergic; Receptors, Histamine; Trachea

The article deals with the possibility of using staphylococcal protein A conjugated with horse-radish peroxidase for the detection of specific IgG-antibodies to ovalbumin in mice by the indirect and competitive EIA techniques. Studies on specifying the parameters of the EIA system for the detection of specific IgG-antibodies are in progress.

Topics: Animals; Antibody Specificity; Hypersensitivity; Immunization; Immunoenzyme Techniques; Immunoglobulin G; Mice; Mice, Inbred CBA; Ovalbumin; Polystyrenes; Staphylococcal Protein A

The effects of enzymic cleavage and perturbing the conformation of the allergenic and antigenic determinants of hens egg white albumin (OA) were examined. Hens egg white extract of a total protein concentration 8.43 g/l was prepared. Isoelectric focusing in sodium dodecyl sulfate and polyacrylamide gel peptide maps for the crude egg white extract showed 26 spots visualized by staining with Coomassie blue. The OA was purified using a TSK-2000 gel filtration chromatography column. The specific allergenic reactivity of the purified OA as measured by RAST inhibition and direct RAST was relatively high: 3 micrograms gave an inhibition of approximately 10%. The cleavage of OA with cyanogen bromide resulted in 4 fractions, all capable of binding specific IgE with the first peak showing the highest inhibition. Thermal denaturation of OA had no direct effect on the antigenic reactivity. RAST inhibition values for the denatured protein were similar to those of the native protein. Carboxymethylation of OA gave a product with only 20% of the inhibition reactivity. Further treatment with trypsin did not abolish the allergenic and antigenic reactivities as shown by RAST inhibition and by deflection of OA line in rocket line immunoelectrophoresis. On the other hand, limited pepsin hydrolysis destroyed the antigenic structure of the molecule. The reactivity of OA is thus relatively stable and could easily be retained making it possible to identify the allergenic determinants of enzymic hydrolysates used for elucidating the antigenic structure of the molecule.

Topics: Allergens; Animals; Antibody Specificity; Cyanogen Bromide; Epitopes; Humans; Hypersensitivity; Immunoelectrophoresis; Immunoglobulin E; Isoelectric Point; Molecular Weight; Ovalbumin; Pepsin A; Peptide Fragments; Protein Denaturation; Radioallergosorbent Test; Trypsin

We previously showed that rats sensitized to egg albumin (EA) respond in vivo intraluminal antigen-challenge with decreased net absorption of water and electrolytes and depletion of mucosal histamine. However, administration of anti-histamines did not prevent the transport abnormalities. The present in vitro studies examined the effect of histamine to alter net ion transport and the ability of diphenhydramine (DPH) and cimetidine (CIM) to block the responses to both histamine and antigen. Control rat jejunum was mounted in Ussing chambers and histamine was added to the serosal side either in the absence or presence of DPH or CIM. In control tissues histamine caused a transient increase in short-circuit current (Isc) in a dose-dependent manner between 10(-5) and 10(-4) M which was blocked by 10(-5) M DPH but was unaffected by CIM in concentrations up to 10(-4) M. There was no response to EA. Jejunum from sensitized rats exposed to EA demonstrated a biphasic Isc response: a rapid transient rise followed by a somewhat less elevated but sustained component. In tissues pre-treated with DPH the initial peak was unaffected but the sustained component was reduced. Our results indicate that H1-receptors mediated the effects of histamine in rat jejunal mucosa but that during intestinal anaphylaxis histamine is responsible for only a portion of the antigen-induced transport abnormalities. Our data also suggest that IgE-mediated reactions in the intestine may involve an interaction between mast cell mediators and enteric nerves.

Topics: Animals; Cimetidine; Diphenhydramine; Histamine; Hypersensitivity; In Vitro Techniques; Intestinal Mucosa; Jejunum; Membrane Potentials; Ovalbumin; Rats; Tetrodotoxin

The ability of azelastine to inhibit IgE-mediated allergic histamine release from the peritoneal mast cells of actively sensitized rats was investigated and compared with selected antiallergic agents. Azelastine added simultaneously with the allergic stimuli (ovalbumin, OA, 10 micrograms/ml + phosphatidylserine, PS, 10 micrograms/ml) or preincubated with cells for 10 min prior to antigen challenge produced similar concentration-dependent inhibition of allergic histamine release. The IC50s (microM) following 10-min preincubation were as follows: azelastine = 4.8; astemizole = 86.3; ketotifen = 112.2; diphenhydramine = 133 and theophylline = 2040.3. At IC50 level azelastine was about 18, 23, 28 and 425 times as effective as astemizole, ketotifen (newer histamine H1-receptor antagonists), diphenhydramine (a traditional H1-receptor antagonist), and theophylline (a phosphodiesterase inhibitor), respectively. Sodium cromoglycate in a concentration range or 1-1000 microM (0 or 10-min preincubation) failed to exert any inhibitory effect. These data showed that among six drugs tested azelastine is the most potent inhibitor of allergic histamine release from rat peritoneal mast cells.

Topics: Animals; Astemizole; Benzimidazoles; Diphenhydramine; Histamine Release; Hypersensitivity; Immunoglobulin E; Ketotifen; Male; Mast Cells; Ovalbumin; Phosphatidylserines; Phthalazines; Pyridazines; Rats; Rats, Inbred Strains; Theophylline

Type-I hypersensitivity reactions were induced in guinea pigs by repeated topical/conjunctival application of fluoresceinyl ovalbumin (FL-OA). The ocular reactivity in early responding animals was maximal between 16 and 25 days and decreased exponentially thereafter. Desensitized eyes responded minimally to compound 48/80 but maximally to histamine. Unilateral sensitization and challenge with FL-OA produced desensitization of the immunized eye, but the reactivity of the contralateral eye persisted. A significant reduction in conjunctival stained mast cells was found in repeatedly challenged eyes. The desensitization therefore was due to a loss of reactive mast cells. Systemic infection with Ascaris suum, after repeated topical challenge with FL-OA had led to desensitization, produced a reappearance of type-I hypersensitivity reactions toward both FL-OA and ascarid antigens.

Topics: Administration, Topical; Animals; Ascariasis; Conjunctival Diseases; Desensitization, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Guinea Pigs; Histamine; Hypersensitivity; Immunization; Mast Cells; Ovalbumin; p-Methoxy-N-methylphenethylamine

We have studied the effects of Ketotifen [Ke] (10(-4)M and 10(-6)M) on two in vitro models of bronchoconstriction: actively sensitized guinea-pig trachea (GPT), and passively sensitized human bronchial muscle (HBM). Experiments were performed on matched pairs of tissues. Cumulative dose response curves [CDRC] for histamine and acetyl choline were constructed, and repeated after pre-incubation with Ke or saline control. The effect of Ke on maximal antigen induced contractions was also studied. Contraction of GPT by histamine and acetyl choline was inhibited by Ke 10(-4)M, though this effect was not apparent at high doses of acetyl choline. Ke 10(-6)M had a weaker inhibitory effect on histamine and acetyl choline induced responses. Contraction of GPT by antigen was unaffected by Ketotifen. In the HBM model, Ke 10(-4)M inhibited acetyl choline and antigen induced responses. Ketotifen, 10(-6)M had an inhibitory effect on acetyl choline induced contractions, though this was small, and not seen at higher agonist doses. Contraction of HBM by antigen was unaffected by Ke 10(-6)M. We were unable to obtain reproducible CD RC's to histamine with HBM. The weaker or absent effects of Ke 10(-6)M, a level close to that obtained in clinical practice, may explain some of the poor results of clinical trials, and suggest that efficacy may be improved by the use of higher doses.

Topics: Acetylcholine; Animals; Antigens; Bronchi; Guinea Pigs; Histamine; Humans; Hypersensitivity; Ketotifen; Male; Muscle Contraction; Muscle, Smooth; Ovalbumin; Trachea

A series of 26 compounds belonging to the chemical class of (1,2,4)triazolo(4,3-a)-quinoxaline-1,4-diones have been investigated for their antiallergic activities in 3 in vitro models of anaphylaxis. Effects of these and other known antiallergic agents on cyclic nucleotide phosphodiesterases (cNUC-PDE) from purified rat mast cells have also been investigated. 18 compounds were potent (I50 less than or equal to 45 microM) inhibitors of antigen-induced release of histamine (AIR) from rat mast cells (RMC), 3 compounds inhibited anti-IgE-induced release of histamine from human basophils (I50 less than or equal to 25 microM) and none of the compounds significantly affected AIR from guinea pig lung slices. 13 of the compounds were more potent than theophylline as inhibitors of cyclic AMP-PDE and/or cyclic GMP-PDE from RMC. Parallel concentration-response curves for the inhibition of cyclic AMP-PDE and cyclic GMP-PDE indicated that these compounds probably interact with enzyme in the same manner. Paired regression analysis of the I50 values for inhibition of AIR and cNUC-PDE from RMC by these compounds revealed no statistically significant correlation between the inhibition of AIR and inhibition of cyclic AMP-PDE or cyclic GMP-PDE. We conclude: (1) some of these compounds are potent inhibitors of immunologic release of histamine from RMC with an in vitro activity profile similar to that of DSCG, and (2) inhibition of cyclic AMP or cyclic GMP hydrolysis by cNUD-PDE by these compounds, DSCG, and 6 known antiallergic agents is not the biochemical mechanism by which they inhibit AIR from RMC.

Topics: 2',3'-Cyclic-Nucleotide Phosphodiesterases; 3',5'-Cyclic-AMP Phosphodiesterases; Animals; Antibodies, Anti-Idiotypic; Antigens; Basophils; Cromolyn Sodium; Depression, Chemical; Guinea Pigs; Histamine H1 Antagonists; Histamine Release; Humans; Hypersensitivity; Immunoglobulin E; Kinetics; Lung; Male; Mast Cells; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Rats; Rats, Inbred Strains; Triazoles

Guinea-pigs sensitised to ovalbumen excrete the antigen in their urine in a therapeutic concentration which prevents anaphylactic death after injection of a challenge dose of the ovalbumen. Sublingual administration of the correct dose of urine from allergic patients also provides therapeutic control of their allergic symptoms. The effective dose is determined by bio-assay. The Neutralisation dose is recognised by disappearance of buccal sensation to the urine. Readministration of salivary, nasal, and sweat secretions from allergic patients onto the conjunctiva also controls allergic symptoms. These procedures provide effective physiological self-defence therapies against allergic challenge in humans.

Topics: Adolescent; Adult; Animals; Antigens; Child; Child, Preschool; Desensitization, Immunologic; Female; Guinea Pigs; Humans; Hypersensitivity; Male; Middle Aged; Nasal Mucosa; Ovalbumin; Saliva; Sweat; Urine

Guinea-pigs were sensitized to ovalbumin by an intraperitoneal injection, followed 3 weeks later by daily aerosol exposure for 4 weeks. Noradrenaline-induced contractions of peripheral lung strips from sensitized and unsensitized guinea-pigs were compared. There were no differences in sensitized or unsensitized strips with respect to maximal tension generated, concentration of noradrenaline for threshold or maximal responses or EC50. These results suggest that any changes in the population of alpha-adrenoceptors consequent upon immunological sensitization did not influence contractile responses to noradrenaline.

Topics: Animals; Antigens; Guinea Pigs; Hypersensitivity; Lung; Male; Muscle Contraction; Muscle, Smooth; Norepinephrine; Ovalbumin

Egg albumin decreased the cyclic AMP content and the protein kinase ratio of sensitized guinea-pig tracheal smooth muscle. The effect preceded the contraction of the tracheal preparation. Antigen challenge increased cyclic AMP phosphodiesterase activity without changing the Km value. In tracheal preparations of desensitized guinea-pigs, the mechanical and metabolic effects of egg albumin were markedly reduced. These results suggest that the effects on the cyclic AMP system in response to immunological challenge might represent an important mechanism modulating the contractile response of guinea-pig tracheal smooth muscle.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Airway Resistance; Animals; Antigens; Cyclic AMP; Female; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Male; Muscle Contraction; Muscle, Smooth; Ovalbumin; Protein Kinases; Respiratory Physiological Phenomena; Respiratory System; Time Factors; Trachea

A dual isolated organ technique comprised of a guinea-pig lung parenchymal strip and a guinea-pig ileum was used to determine if slow reacting substance of anaphylaxis (SRS-A) is released from parenchyma during contractions evoked by antigen (ovalbumin) or by ionophore (A23187). An immunologically sensitized parenchyma served as the primary target organ for ovalbumin and either a sensitized or unsensitized parenchyma was the target tissue for A23187; an unsensitized ileum functioned as the assay organ. In the presence of pyrilamine and indomethacin, ovalbumin or A23187 produced contractions of the parenchyma and concomitantly caused release of SRS-A from the lung strip which was indicated by a contraction of the ileum. The ileal response was antagonized by FPL 55712, whereas the parenchyma contractions were unaffected. Additional experiments were conducted in which parenchyma was contracted with histamine. At the height of the histamine contraction, the bathing fluid surrounding the parenchyma was removed and assayed on a pyrilamine-treated ileum. SRS-A was not detected, indicating that SRS-A release from parenchyma is not a function of tissue contraction per se, but is related to the antigen- and ionophore-induced contractions. To explain the lack of effect of FPL 55712 on parenchymal contractions to antigen or ionophore, we compared the degree of antagonism produced by FPL 55712 on SRS-A contraction of parenchyma and ileum. These experiments indicated the possibility that at least two different classes of SRS-A receptors exist and that those in the ileum and lung differ.

Topics: Animals; Calcimycin; Chromones; Guinea Pigs; Histamine; Hypersensitivity; In Vitro Techniques; Lung; Male; Muscle Contraction; Muscle, Smooth; Ovalbumin; SRS-A; Time Factors

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Guinea Pigs; Hypersensitivity; In Vitro Techniques; Indomethacin; Kinetics; Lung; Male; Microsomes; Ovalbumin; Prostaglandin-Endoperoxide Synthases; Thromboxane B2; Time Factors

Leukotriene (SRS) appears to contribute to the prolonged phase of the tracheal contraction induced by egg albumin, indomethacin and tartrazine as this response was effectively antagonised by FPL 55712. Egg albumin, indomethacin and tartrazine reduced the cAMP level of guinea pig trachea pretreated with adrenergic, cholinergic, histaminergic and 5-hydroxytryptaminergic antagonists. Egg albumin, but not the other agents, caused a transient increase of the cGMP content. The metabolic and contractile effects were blocked by FPL 55712. It is suggested that the cAMP reduction caused by egg albumin, indomethacin and tartrazine might promote contractions ot the tracheal muscle.

Topics: Animals; Chromones; Female; Guinea Pigs; Hypersensitivity; Indomethacin; Male; Muscle Contraction; Muscle, Smooth; Nucleotides, Cyclic; Ovalbumin; SRS-A; Tartrazine; Trachea

The effect of a water-soluble fraction (CEF) that was prepared from an extract of Corynebacterium equi on primary reaginic antibody formation was studied in Balb/c mice. Mice were immunized with a hapten carrier (DNP-OVA) and received intraperitoneal injections of CEF 7 and 2 days prior to, or 2 and 7 days after the immunization. PCA titers of both antihapten (DNP) and anticarrier (OVA) antibodies of IgE class were reduced significantly by the CEF treatment. Evidence was presented in adoptive transfer experiments that the number of IgE-producing cells in the CEF-treated mice was lower than that of controls. Suppression of IgG1 anti-DNP antibody formation was also achieved by the CEF treatment. Formation of IgG1 anti-OVA antibodies, however, was not suppressed significantly by the treatment. The suppressive activities of CEF were shown to be dose-dependent, but timing of CEF administration did not appear critical.

Topics: Aerobiosis; Animals; Antibody Formation; Antibody-Producing Cells; Carrier Proteins; Chemical Fractionation; Corynebacterium; Dinitrobenzenes; Epitopes; Haptens; Hypersensitivity; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Reagins; Time Factors

Topics: Airway Resistance; Animals; Cyclooxygenase Inhibitors; Dose-Response Relationship, Drug; Female; Guinea Pigs; Histamine; Hypersensitivity; Lipoxygenase Inhibitors; Ovalbumin; Respiratory System

Enhancement of allergic tracheal constriction and contraction of the trachea by AA in the presence of cyclooxygenase inhibitors may reflect diversion of AA metabolism from the cyclooxygenase pathway.

Topics: Airway Resistance; Animals; Cyclooxygenase Inhibitors; Guinea Pigs; Hypersensitivity; Indomethacin; Lipoxygenase Inhibitors; Male; Meclofenamic Acid; ortho-Aminobenzoates; Ovalbumin; Prostaglandins E; Pyrazoles; Trachea

Topics: Animals; Antibody Specificity; Hookworm Infections; Hypersensitivity; Immunization, Passive; Immunoglobulin E; Mast Cells; Nippostrongylus; Ovalbumin; Rats; Receptors, Immunologic; Skin Tests

Topics: Animals; Atropine; Guinea Pigs; Histamine; Hypersensitivity; Isoproterenol; Lung; Male; Ovalbumin; Phenylephrine; Practolol; Prostaglandins; Sotalol; SRS-A; Thromboxane A2; Thromboxanes

Passive cutaneous anaphylaxis (PCA) mediated in rats by IgE-like antibodies against egg albumin or the benzylpenicilloyl determinant was inhibited in a dose-dependent manner by intravenous treatment with Y-12,141; the ED50 was 0.09--0.2 mg/kg. The inhibitory effect of Y-12,141 ON PCA was about 5 times as potent as that of disodium cromoglycate (DSCG). Oral treatment with Y-12,141 resulted in the inhibition of PCA, showing an ED50 of 2.5 mg/kg. This action of Y-12,141 on PCA was considered to be due to the inhibition of the release of allergic mediatros from mast cells in a manner similar to DSCG. The results suggest that Y-12,141 may have an anti-allergic activity.

Topics: Animals; Azoles; Chromones; Cromolyn Sodium; Dose-Response Relationship, Immunologic; Female; Histamine Antagonists; Histamine Release; Hypersensitivity; Immunoglobulin E; Male; Mice; Mice, Inbred ICR; Ovalbumin; Passive Cutaneous Anaphylaxis; Penicillin G; Rats; Tetrazoles

Administration of stable conjugates prepared by coupling protein antigens such as ovalbumin or antigen E of ragweed extract to the synthetic random copolymer of D-glutamic acid and D-lysine (D-GL) is effective in inducing a state of long-lasting, antigen-specific immunological tolerance in experimental animals. A striking aspect of the tolerance induced by protein-D-GL conjugates is the remarkable selectivity of the tolerance for antibody responses of the IgE class. Protein-D-GL conjugates of either type were capable of inducing such tolerance both in unsensitized and in previously sensitized animals when administered in appropriate doses. Comparable doses of unconjugated proteins were likewise capable of suppressing IgE antibody production, although the duration of suppression in these cases was significantly less than that observed with protein-D-GL conjugates. If such conjugates act in man as they do in experimental animals, they could be of great value as therapeutic agents in selectively diminishing IgE antibody production while sparing antibody production in the IgG class.

Topics: Animals; Antibody Formation; Antigens; Hypersensitivity; Immunoglobulin E; Immunotherapy; Mice; Mice, Inbred Strains; Ovalbumin; Plants; Radioimmunoassay

Topics: Animals; Antibody Formation; Eye; Hypersensitivity; Ovalbumin; Rabbits; Uveitis

Six non-steroidal anti-inflammatory drugs, metiazinic and niflumic acids, iburofen, indometacin, ketoprofen and phenylbutazone, were tested on two anaphylactic skin reactions to ovalbumin: the passive reaction in the rat and active anaphylaxis in the guinea pig. In the rat, only niflumic and metiazinic acids reduced very slightly the reaction. In the guinea pig, ibuprofen, phenylbutazone, metiazinic acid and ketoprofen had good activity. The results are discussed.

Topics: Animals; Anti-Inflammatory Agents; Guinea Pigs; Hypersensitivity; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Skin Diseases

Topics: Anaphylaxis; Animals; Drug Evaluation; Guinea Pigs; Histamine H1 Antagonists; Hypersensitivity; Male; Mast Cells; Ovalbumin; Piperazines

Topics: Animals; Antigens; Binding, Competitive; Guinea Pigs; Histamine Release; Hypersensitivity; Isoproterenol; Lung; Ovalbumin; Rats; Receptors, Adrenergic; Receptors, Adrenergic, beta; Skin; Skin Physiological Phenomena

Topics: Complement C3; Cystic Fibrosis; Electrophoresis, Agar Gel; Fluorescent Antibody Technique; Humans; Hypersensitivity; Immunoglobulin A, Secretory; Immunoglobulin E; Ovalbumin; Precipitins; Saliva; Serum Albumin, Bovine; Sputum

Topics: Administration, Intranasal; Aerosols; Animals; Antibodies; Antigens; Bronchi; Bronchography; Cattle; Constriction, Pathologic; Disease Models, Animal; Dose-Response Relationship, Immunologic; Guinea Pigs; Hypersensitivity; Lung; Lung Compliance; Lung Volume Measurements; Ovalbumin; Pyrilamine; Respiration; Serum Albumin, Bovine; Tracheotomy

Topics: Animals; Antibody Specificity; Cell Line; Hot Temperature; Hybrid Cells; Hypersensitivity; Immunoglobulin E; Male; Mice; Myeloma Proteins; Ovalbumin; Protein Denaturation; Rats; Skin Tests; Spleen

PRD-92-Ea [5,5-Dimethyl-11-oxo-5H, 11H-(2) benzopyrano (4,3-g) (1) benzopyran-9-carboxylic acid ethanolamine], was an active antiallergic compound in rat and monkey experimental models of immediate hypersensitivity. It inhibited, in a dose-dependent manner, the rat PCA reaction after both intravenous and oral administration. It also inhibited the degranulation of rat peritoneal mast cells after antigenic challenge. PRD-92-Ea was also active in preventing bronchoconstriction in Ascaris-sensitive Rhesus monkeys after intravenous, topical and oral administration. Using chopped monkey tissues, it was found that PRD-92-Ea prevented histamine release from the respiratory mast cells, but not from the cutaneous mast cells. No reason for this dichotomy of effect is known. PRD-92-Ea showed antagonistic activity against the allergic mediators released from mast cells. In order of decreasing potency it was active against SRS-A (monkey lung), PGF2alpha, PGE2, serotonin, bradykinin and histamine. Apart from its antiallergic effects PRD-92-Ea had no other significant pharmacological activity.

Topics: Animals; Asthma; Benzopyrans; Cromolyn Sodium; Disease Models, Animal; Haplorhini; Histamine Release; Hypersensitivity; Isoproterenol; Macaca mulatta; Male; Mast Cells; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats

A number of 2-cyanoindan-1,3-diones and 3-cyano-4-hydroxycoumarins have been prepared and assessed for potential antiallergy activity as measured by their ability to inhibit passive cutaneous anaphylaxis in the rat, mediated by rat serum containing antigen specific IgE. The structural requirements for activity were similar not only for both series of compounds but also for the analogous 2-nitroindan-1,3-diones and 4-hydroxy-3-nitrocoumarins previously reported. The most active compounds were 2-cyano-5,6-diethylindan-1,3-dione (4e) and 3-cyano-6,7-diethyl-4-hydroxycoumarin (11h).

Topics: 4-Hydroxycoumarins; Animals; Cromolyn Sodium; Hypersensitivity; Immunoglobulin E; Indans; Indenes; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Structure-Activity Relationship

In this article a study on the influence on bone-marrow eosinophilia in the mouse of a single injection of various concentrations of egg-white and incomplete Freund's adjuvant, together and separately, is reported. The combination of egg-white (1 per cent) and adjuvant gave a peak value of 158.3% eosinophil leukocytes on the 14th day after the injection; separately, neither one led to divergence from the results in untreated mice. The combination gave the greatest effect at 1.0 per cent and 0.1 per cent egg-white, a moderate effect with 10 per cent and 0.01 per cent, and negligible effects with 0.001 per cent. The interpretation of the results is discussed, with special attention to the relationship with the atopy syndrome in man.

Topics: Animals; Bone Marrow; Bone Marrow Cells; Bone Marrow Examination; Eosinophilia; Eosinophils; Freund's Adjuvant; Hypersensitivity; Leukocyte Count; Mice; Ovalbumin; Probability

Eighty-eight asthmatic children aged six to 16 years received monovalent influenza A/New Jersey/76 virus vaccines. Forty-one of these children were given skin tests for allergy to eggs and vaccines, and 57 were given pulmonary function tests before and after immunization. Only four children reacted to the vaccines in the skin tests, with three of these children reacting to only one of four test preparations. Only two of the four children showed a correlation between reactivity to vaccine and allergy to egg antigens. No significant changes in pulmonary function were demonstrated.

Topics: Adolescent; Asthma; Child; Egg White; Forced Expiratory Volume; Humans; Hypersensitivity; Influenza A virus; Influenza Vaccines; Maximal Midexpiratory Flow Rate; New Jersey; Ovalbumin; Respiratory Function Tests; Skin Tests

Topics: Animals; Blood Pressure; Female; Hypersensitivity; Hypersplenism; Hypertension, Portal; Injections, Intramuscular; Liver; Male; Organ Size; Ovalbumin; Portal System; Rabbits; Spleen

Systemic treatment with a heterologous anti-T cell serum of guinea pigs immunized with EA in IFA markedly suppressed CBH reactivity to specific antigen and T cell mitogens, as judged by gross reactivity, histology, and skin histamine. The antiserum produced a marked drop in circulating lymphocytes, mainly at the expense of T cells, as indicated by the ability of surviving lymphocytes to rosette with rabbit RBC. It was postulated that the suppression of CBH reactivity is due to the depletion of T cells, which would have released a factor chemotactic for basophils. The data therefore provide further evidence that cutaneous reactions rich in basophils are primarily dependent on a population of T cells.

Topics: Animals; Antilymphocyte Serum; Basophils; Concanavalin A; Female; Guinea Pigs; Histamine Release; Hypersensitivity; Immune Adherence Reaction; Lectins; Leukocyte Count; Lipopolysaccharides; Mitogens; Ovalbumin; Skin Tests; T-Lymphocytes

Cats, once thought to possess little immunological responsiveness, have recently been shown to manifest a wide range of immunologic reactions. Sensitization of cats with ovalbumin rendered the animals capable, after a latency period of 40 to 60 days, of anaphylactic shock upon intravenous challenge. Physiologically the response is characterized by bronchoconstriction as evidenced by an increase in airways resistance. This response was ameliorated by intravenous administration of either aminophylline or isoproterenol. Other reactions elicited upon challenge were dyspnea, cyanosis, and in some instances death. The parameters of the allergic response corresponded with those reported in previous studies of other experimental models of allergic bronchoconstriction.

Topics: Airway Resistance; Aminophylline; Anesthesia; Animals; Bronchi; Cats; Female; Hypersensitivity; Isoproterenol; Male; Ovalbumin; Time Factors

Topics: Animals; Arachidonic Acids; Glutathione; Guinea Pigs; Hypersensitivity; Lung; Mixed Function Oxygenases; Ovalbumin; Prostaglandin-Endoperoxide Synthases; Prostaglandins

The syntheses of trans-3-(4-oxo-4H-1-benzypyran-3)acrylic acid and a number of analogs shown to be highly active in antiallergic bioassays are described. These compounds are of possible value in the treatment of asthma. The structural requirements for biological activity are discussed with reference to the type of the substituents on the chromone ring or positions of linkage of the acrylic acid on the pyrone ring.

Topics: Acrylates; Anaphylaxis; Animals; Bacillus; Benzopyrans; Dose-Response Relationship, Drug; Hypersensitivity; Immune Sera; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Stereoisomerism; Structure-Activity Relationship

Alterations in pulmonary conductance, dynamic compliance, respiratory frequency, minute volume, mean arterial pressure, pulse rate, relaxation volume-to-dry weight ratio, and wet-to-dry weight ratio resulting from antigen infusion in sensitized guinea pigs was examined with and without atropine treatment. In untreated animals 3 min after antigen infusion there were significant decreases in dynamic compliance and pulmonary conductance with an increase in relaxation volume-to-dry weight ratio while other parameters were not altered. In atropine-treated animals antigen infusion resulted in a decreased dynamic compliance and an increased relaxation volume-to-dry weight ratio but no significant change in pulmonary conductance. This suggests that the alterations in large and central airway tone resulting from antigen infusion are mediated predominantly by secondary cholinergic mechanisms while peripheral airway effects are mainly noncholinergic.

Topics: Animals; Antigens; Asthma; Atropine; Guinea Pigs; Hypersensitivity; Infusions, Parenteral; Lung; Lung Compliance; Male; Organ Size; Ovalbumin; Oxygen Consumption; Parasympathetic Nervous System; Pulmonary Ventilation; Respiration

Topics: Animals; Antibody Formation; Bordetella pertussis; Hypersensitivity; Ovalbumin; Passive Cutaneous Anaphylaxis; Quinolines; Rats; Structure-Activity Relationship

Topics: Anaphylaxis; Animals; Antigens, Bacterial; Bordetella pertussis; Cromolyn Sodium; Dose-Response Relationship, Drug; Eye; Guinea Pigs; Histamine Release; Hot Temperature; Hypersensitivity; Immune Sera; Immunization; Immunoglobulin E; Isoantibodies; Lung; Male; Mercaptoethanol; Ovalbumin; Passive Cutaneous Anaphylaxis

Topics: Animals; Antigen-Antibody Reactions; Histamine H1 Antagonists; Histamine Release; Hypersensitivity; Indenes; Male; Nitro Compounds; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats

Topics: Adjuvants, Immunologic; Animals; Antibodies; Body Temperature; Fluorescent Antibody Technique; Hydrocarbons; Hypersensitivity; Immunodiffusion; Liver; Lymph Nodes; Ovalbumin; Rats; Serotonin; Spleen

Topics: Acetylcholine; Antibody Formation; Asthma; Bradykinin; Bronchi; Carotid Sinus; Histamine; Hypersensitivity; Ovalbumin; Pilocarpine; Pressoreceptors; Serotonin; Spirometry; Vagotomy; Vagus Nerve

Topics: Animals; Antigen-Antibody Reactions; Antigens; Basophils; Chickens; Erythrocytes; Female; Formaldehyde; Guinea Pigs; Hypersensitivity; Immunity, Cellular; Immunization; Macrophages; Ovalbumin; Sheep; Skin; Skin Tests

Topics: Air; Anaphylaxis; Animals; Aorta; Arachis; Blood Pressure; Dextrans; Drug Hypersensitivity; Emulsions; Female; Guinea Pigs; Histamine Release; Hypersensitivity; Iron-Dextran Complex; Lung; Male; Muscle Contraction; Oils; Ovalbumin; Perfusion; Polystyrenes; Pressure; Prostaglandins; Pulmonary Artery; Rabbits; Rats; Stimulation, Chemical; Suspensions; Time Factors

Quantitation of soluble residual host protein in chicken embryo-derived vaccines was performed rapidly, economically, and accurately by radial immunodiffusion.

Topics: Animals; Chick Embryo; Humans; Hypersensitivity; Immune Sera; Immunodiffusion; Immunoelectrophoresis; Influenza Vaccines; Measles Vaccine; Methods; Ovalbumin; Proteins; Rabbits; Smallpox Vaccine; Viral Vaccines; Yellow Fever

Topics: Animals; Antibody Formation; Antigens; Binding Sites; Coloring Agents; Female; Hookworm Infections; Hypersensitivity; Immunization, Passive; Immunoglobulin E; Injections, Intradermal; Injections, Intraperitoneal; Kinetics; Mast Cells; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Skin Tests

Topics: Animals; Antigens; Beta-Globulins; Columbidae; Cross Reactions; Egg Yolk; Feces; Female; gamma-Globulins; Humans; Hypersensitivity; Immunoelectrophoresis; Ovalbumin; Plasma; Pneumonia; Proteins; Rabbits; Serum Albumin

Topics: Animals; Antigen-Antibody Reactions; Haplorhini; Hypersensitivity; Ovalbumin; Periodontal Diseases

Topics: Acid Phosphatase; Amyloidosis; Animals; Antigens; Caseins; Hypersensitivity; Male; Mice; Ovalbumin; Serum Albumin

Topics: Animals; Cholinesterases; Glutathione; Hypersensitivity; Keratitis; Ovalbumin; Rabbits

Topics: Anaphylaxis; Animals; Cat Diseases; Cats; Fibrinogen; Freund's Adjuvant; gamma-Globulins; Hypersensitivity; Ovalbumin; Serum Albumin, Bovine

Topics: Adenosine Triphosphate; Aerosols; Aldehyde-Lyases; Anaphylaxis; Animals; Antigens; Glycolysis; Guinea Pigs; Hexosephosphates; Hypersensitivity; L-Lactate Dehydrogenase; Lung; Mitochondria; Ovalbumin; Time Factors

Topics: Animals; Antilymphocyte Serum; Horses; Hypersensitivity; Injections; Injections, Intramuscular; Injections, Intravenous; Ovalbumin; Rabbits; Uveitis; Vitreous Body

Topics: Animals; Antigen-Antibody Reactions; Guinea Pigs; Heart Rate; Hypersensitivity; In Vitro Techniques; Lymph Nodes; Mesentery; Ovalbumin; Splenectomy

Topics: Animals; Bronchi; Epithelium; Guinea Pigs; Hypersensitivity; Lung; Nitrous Oxide; Organ Size; Ovalbumin; Respiratory Hypersensitivity; Skin Tests

Topics: Animals; Antibody Formation; Antigens; Ascaris; Bordetella pertussis; Cellulose; Chromatography; Guinea Pigs; Hemocyanins; Histamine Release; Hypersensitivity; Immune Sera; Immunoelectrophoresis; Immunoglobulin E; Injections, Subcutaneous; Ovalbumin; Passive Cutaneous Anaphylaxis; Proteins; Rats; Skin Tests; Tissue Extracts; Vaccines

Topics: Anaphylaxis; Animals; Bile Acids and Salts; Bradykinin; Carboxy-Lyases; Esophagus; Guinea Pigs; Histamine; Histamine H1 Antagonists; Hypersensitivity; In Vitro Techniques; Muscle Contraction; Ovalbumin; Quinolines; Serotonin

Topics: Adjuvants, Immunologic; Animals; Antibody Formation; Antigens; Cornea; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Hypersensitivity; Immunity; Injections; Lymph Nodes; Male; Methods; Mycobacterium; Ovalbumin; Skin Tests

Topics: Allergens; Amides; Aminopeptidases; Animals; Blood; Chromatography, Gel; Dust; Feathers; Glucose; Glycoproteins; Glycyrrhiza; Humans; Hypersensitivity; Kallidin; Leucine; Lysine; Naphthalenes; Ovalbumin; Plants, Medicinal; Pollen; Ribose; Skin Tests

Topics: Animals; Antigens; Arthritis; Freund's Adjuvant; Hypersensitivity; Ovalbumin; Rats; Skin Tests

Topics: Animals; Antigen-Antibody Reactions; Antigens; Ascitic Fluid; Blood; Cattle; Dialysis; Female; Histamine Release; Horses; Humans; Hypersensitivity; Immune Sera; Macromolecular Substances; Mast Cells; Ovalbumin; Rabbits; Rats

Topics: Anaphylaxis; Animals; Antigens; Biological Assay; Female; Guinea Pigs; Histamine; Histamine Release; Hypersensitivity; Ileum; Immune Sera; Liver; Lung; Male; Muscle Contraction; Ovalbumin; Rabbits; Stimulation, Chemical; Time Factors

Topics: Animals; Ear, Inner; Guinea Pigs; Hypersensitivity; Otitis Media; Ovalbumin; Proteus

Topics: Animals; Conjunctivitis; Eosinophilia; Hypersensitivity; In Vitro Techniques; Male; Ovalbumin; Rabbits

Topics: Animals; Animals, Newborn; Antigen-Antibody Reactions; Antigens; Antilymphocyte Serum; Complement Fixation Tests; Hypersensitivity; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immune Sera; Leukocyte Count; Lymphocytes; Lymphoid Tissue; Mice; Ovalbumin; Research; Skin Tests; Thymectomy; Tuberculin Test

Topics: Adolescent; Adult; Albumins; Animals; Antibodies; Antigen-Antibody Reactions; Cardiovascular Diseases; Cattle; Child; Collagen Diseases; Communicable Diseases; Diabetes Mellitus; Diabetes Mellitus, Type 1; Female; gamma-Globulins; Gastrointestinal Diseases; Hematologic Diseases; Humans; Hypersensitivity; Infant; Infant, Newborn; Insulin; Iodine Isotopes; Lactalbumin; Maternal-Fetal Exchange; Milk; Nervous System Diseases; Ovalbumin; Pregnancy; Respiratory Tract Diseases; Serum Albumin; Serum Albumin, Bovine; Skin Tests; Viral Vaccines

Topics: Animals; Antibody Formation; Antigen-Antibody Reactions; Antigens; Bone Marrow Cells; Cattle; gamma-Globulins; Guinea Pigs; Hypersensitivity; Hypersensitivity, Delayed; Immunosuppressive Agents; Ovalbumin; Radiation Effects; Research; Serum Albumin; Serum Albumin, Bovine; Skin Transplantation; Spleen; Transplantation Immunology

Topics: Animals; Antigen-Antibody Reactions; Guinea Pigs; Hypersensitivity; Injections; Injections, Intravenous; Ovalbumin; Passive Cutaneous Anaphylaxis; Research

Topics: Adipose Tissue; Allergy and Immunology; Animals; Beta-Globulins; Guinea Pigs; Hypersensitivity; Hypersensitivity, Delayed; Macrophages; Muscles; Necrosis; Ovalbumin; Pathology; Research; Skin Tests

Topics: Adjuvants, Immunologic; Amino Acids; Amino Sugars; Antigen-Antibody Reactions; Biological Assay; Chemical Phenomena; Chemistry; Cornea; Encephalomyelitis; Glycolipids; Granuloma; Hypersensitivity; Mycobacterium; Mycobacterium bovis; Mycobacterium tuberculosis; Ovalbumin; Precipitin Tests; Research; Waxes

Topics: gamma-Globulins; Hypersensitivity; Hypersensitivity, Delayed; Immunity; Mechlorethamine; Mercaptopurine; Ovalbumin; Rabbits; Research; Skin Tests

Topics: Anaphylaxis; Animals; Capillaries; Diphenhydramine; Electrons; Ferritins; Guinea Pigs; Hypersensitivity; Immune System Phenomena; Lung; Microscopy; Microscopy, Electron; Ovalbumin; Pharmacology; Research; Thrombosis

Topics: Cats; Coloring Agents; Eye Diseases; Humans; Hypersensitivity; Microscopy; Ovalbumin; Pathology; Research; Retinal Vessels; Staining and Labeling; Vitreous Body

Topics: Anaphylaxis; Dextrans; Genetics; Hypersensitivity; Immune System Diseases; Ovalbumin; Rats; Research; Toxicology

Topics: Animals; Guinea Pigs; Hypersensitivity; Mice; Nitrobenzenes; Ovalbumin; Research; Serum Albumin; Skin Tests; Tuberculin Test

Topics: 4-Aminobenzoic Acid; Aminobenzoates; Animals; Antibodies; Antibody Formation; Antigens; BCG Vaccine; gamma-Globulins; Guinea Pigs; Hypersensitivity; Hypersensitivity, Delayed; Mycobacterium bovis; Ovalbumin; Proteins; Research; Skin Tests; Trichloroacetic Acid; Vaccination

Topics: Allografts; Animals; Diphtheria Toxoid; Guinea Pigs; Hypersensitivity; Hypersensitivity, Delayed; In Vitro Techniques; Ovalbumin; Research; Skin Transplantation; Transplantation Immunology; Transplantation, Homologous; Tuberculin Test

Topics: Anaphylaxis; Dextrans; Egg White; Hypersensitivity; Inflammation; Ovalbumin; Pharmacology; Rats; Research; Toxicology; Trypsin Inhibitors; Vascular Diseases

Some characteristics of inhibition of cell migration induced in tissue culture by the addition of specific antigen were studied. The following characteristics were found to be shared by this type of cellular hypersensitivity and delayed cutaneous sensitivity: 1. Specificity for the carrier moiety of haptene protein conjugates. The picryl protein conjugate used to sensitize guinea pigs inhibited migration of monocytic cells from these animals. Other picrylated proteins produced little inhibition. 2. Enhancement by mycobacterial adjuvants. Incorporation of tubercle bacilli with picrylated proteins in adjuvant-antigen emulsions stimulated the development of this cellular hypersensitivity to antigen. 3. Independence of circulating antibody. In contrast to cellular hypersensitivity, serum antibody (a) reacted with any of a number of picrylated proteins, (b) developed well in the absence of mycobacterial adjuvant, and (c) persisted in unchanged titer for 5 weeks in animals sensitized with saline solutions of antigen. During this time cellular hypersensitivity decreased remarkably. The in vitro system described provides a direct method to measure cell-antigen interaction and permits study of an aspect of the immune response not mediated by humoral antibody. The relation of cellular hypersensitivity to antibody formation and delayed hypersensitivity is discussed.

Topics: Adjuvants, Immunologic; Animals; Antibodies; Antibody Formation; Antigens; Cell Movement; Chemical Precipitation; Guinea Pigs; Haptens; Hypersensitivity; Hypersensitivity, Delayed; Immunization; In Vitro Techniques; Lung; Ovalbumin; Picrates; Proteins; Research; Skin; Spleen; Tissue Culture Techniques; Tissue Extracts; Vaccination

Topics: Adsorption; Animals; Antibodies; Chemistry Techniques, Analytical; Fluorescent Antibody Technique; gamma-Globulins; Globulins; Hypersensitivity; Ovalbumin; Precipitins; Rabbits; Reagins; Research; Silk; Skin; Textiles

Topics: Animals; Guinea Pigs; Hypersensitivity; Injections; Meniere Disease; Ovalbumin; Pharmacology; Research; Sodium Chloride; Tympanic Membrane

Topics: Adjuvants, Immunologic; Animals; Antigens; Freund's Adjuvant; Guinea Pigs; Hypersensitivity; Hypersensitivity, Delayed; Injections, Intramuscular; Injections, Intravenous; Ovalbumin; Research; Skin Tests

Topics: Animals; Guinea Pigs; Histamine; Hypersensitivity; Injections; Injections, Subcutaneous; Ovalbumin; Pharmacology; Research; Respiratory System; Toxicology

Topics: Administration, Cutaneous; Animals; Antigen-Antibody Reactions; Dermatitis, Atopic; Hypersensitivity; Hypersensitivity, Immediate; Mast Cells; Mice; Ovalbumin; Research

Topics: Antigen-Antibody Reactions; Colitis; Fluorescence; Fluorescent Antibody Technique; Hypersensitivity; Microscopy; Microscopy, Fluorescence; Ovalbumin; Rabbits; Research; Toxicology

Topics: Abscess; Candida; Hypersensitivity; Kidney Diseases; Liver Abscess; Lung Abscess; Ovalbumin; Pathology; Rats; Research; Sepsis; Staphylococcal Infections