ovalbumin and Airway-Obstruction

Topics: Airway Obstruction; Animals; Asthma; Disease Models, Animal; Disease Progression; Eosinophils; Humans; Immunization; Nasal Provocation Tests; Ovalbumin

Asthma is a chronic inflammatory disease of the airways of the lungs. Pomalidomide (POM) a therapy for multiple myeloma has been stated to have an anti-inflammatory effect. The main goal of the present study was to assess its possible effect on airway contraction and inflammation in a rat model of ovalbumin-induced asthma. Different groups of rats received saline or pomalidomide (0.4, 0.8 mg/kg) or dexamethasone (0.6 mg/kg). The asthma was induced by ovalbumin (OVA). Trachea contraction was assayed by organ bath system. Airway histology was assessed using hematoxylin and eosin method. Serum Tumor necrosis factor alpha (TNF-α) level was analyzed by Enzyme-Linked Immunosorbent Assay and Platelet-derived growth factor (PDGFα) Gene expressions were evaluated by Real-time PCR. Pomalidomide prevented ovalbumin-induced airway contraction and histopathological damage. In addition serum, TNF-α level was significantly (p<0.05) decreased in POM treated animals compared to control (asthmatic animals that received POM vehicle). Results indicate that POM prevented the PDGF expression induced by ovalbumin. In conclusion, we found that pomalidomide ameliorated the symptoms, histopathological changes and inflammatory markers induced by ovalbumin in asthmatic rats and these effects might be related to its anti-inflammatory properties.

Topics: Airway Obstruction; Allergens; Animals; Anti-Inflammatory Agents; Asthma; Disease Models, Animal; Humans; Lung; Male; Ovalbumin; Platelet-Derived Growth Factor; Rats; Rats, Wistar; Thalidomide; Tumor Necrosis Factor-alpha

The two life-threatening signs of anaphylactic shock (AS) are severe arterial hypotension and bronchospasm. Guidelines recommend epinephrine as first-line treatment. Arginine vasopressin (AVP) has been proposed as an alternative if epinephrine does not correct arterial hypotension. These two drugs may have beneficial, neutral or deleterious effects on airflow either directly or by modifying factors that regulate vasodilatation and/or edema in the bronchial wall.. To compare the effects of epinephrine and AVP on airflow and airway leakage in a rat model of AS.. Thirty-two ovalbumin-sensitized rats were randomized into four groups: control (CON), AS without treatment (OVA), AS treated with epinephrine (EPI), and AS treated with AVP (AVP). Mean arterial pressure (MAP), respiratory resistance and elastance and microvascular leakage in the airways were measured.. All OVA rats died within 20 minutes following ovalbumin injection. Ovalbumin induced severe arterial hypotension and airway obstruction (221 ± 36 hPa.s.L. Epinephrine was superior to AVP for alleviating the airway response in a rat model of AS. When bronchospasm and severe arterial hypotension are present during AS, epinephrine should be the drug of choice.

Topics: Airway Obstruction; Anaphylaxis; Animals; Arterial Pressure; Bronchial Spasm; Capillary Leak Syndrome; Epinephrine; Hypotension; Neurophysins; Ovalbumin; Protein Precursors; Rats; Respiratory System; Vasopressins

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

Asthma is a highly prevalent chronic inflammatory lung disease characterized by airway hyperresponsiveness to allergens, airway edema, and increased mucus secretion. Such mucus can be liquefied by recombinant human deoxyribonuclease (rhDNase), in which efficacy of rhDNase has been well documented in patients with cystic fibrosis, but little studied in asthma. In the present study, we investigated whether rhDNase intranasal administration improved inflammation and pulmonary function in an experimental model of asthma.. Mice were sensitized by two subcutaneous injections of ovalbumin (OVA), on days 0 and 7, followed by three intranasal challenges with OVA on days 14, 15, and 16. A control group, replacing OVA by DPBS, was included. On days 15 and 16, after 2 hours of OVA challenge, mice received 1 mg/mL of intranasal rhDNase.. We showed that rhDNase decreased significantly the airway resistance and reduced EETs formation and globet cells hyperplasia.. Our results suggest that extracellular DNA in mucus play a role in lower airways obstruction in OVA asthma protocol and that the treatment with rhDNase improved lung function and DNA extracellular traps, with no direct cellular anti-inflammatory effects.

Topics: Administration, Intranasal; Airway Obstruction; Airway Resistance; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Deoxyribonucleases; Disease Models, Animal; DNA; Extracellular Traps; Female; Humans; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Recombinant Proteins

The ethanol extract of KOTMIN13, composed of Inula japonica Flowers, Trichosanthes kirilowii Semen, Peucedanum praeruptorum Radix, and Allium macrostemon Bulbs, was investigated for its anti-asthmatic and anti-allergic activities.. The anti-asthmatic effects of KOTMIN13 were evaluated on ovalbumin (OVA)-induced murine asthma model. Anti-allergic properties of KOTMIN13 in bone-marrow derived mast cells (BMMC) and passive cutaneous anaphylaxis (PCA) in vivo were also examined.. In asthma model, KOTMIN13 effectively suppressed airway hyperresponsiveness induced by aerosolized methacholine when compared to the levels of OVA-induced mice. KOTMIN13 treatment reduced the total leukocytes, eosinophil percentage, and Th2 cytokines in the bronchoalveolar lavage fluids in OVA-induced mice. The increased levels of eotaxin and Th2 cytokines in the lung as well as serum IgE were decreased by KOTMIN13. The histological analysis shows that the increased inflammatory cell infiltration and mucus secretion were also reduced. In addition, the degranulation and leukotriene C4 production were inhibited in BMMC with IC50 values of 3.9 μg/ml and 1.7 μg/ml, respectively. Furthermore, KOTMIN13 treatment attenuated mast-mediated PCA reaction.. These results demonstrate that KOTMIN13 has anti-asthmatic and anti-allergic effects in vivo and in vitro models.

Topics: Airway Obstruction; Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Female; Herbal Medicine; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts

Atopic, obese asthmatics exhibit airway obstruction with variable degrees of eosinophilic airway inflammation. We previously reported that mice obese as a result of a genetic deficiency in either leptin (ob/ob mice) or the long isoform of the leptin receptor (db/db mice) exhibit enhanced airway obstruction in the presence of decreased numbers of bronchoalveolar lavage fluid (BALF) eosinophils compared with lean, wild-type mice following antigen (ovalbumin; OVA) sensitization and challenge. To determine whether the genetic modality of obesity induction influences the development of OVA-induced airway obstruction and OVA-induced pulmonary inflammation, we examined indices of these sequelae in mice obese as a result of a genetic deficiency in carboxypeptidase E, an enzyme that processes prohormones and proneuropeptides involved in satiety and energy expenditure (Cpe(fat) mice). Accordingly, Cpe(fat) and lean, wild-type (C57BL/6) mice were sensitized to OVA and then challenged with either aerosolized PBS or OVA. Compared with genotype-matched, OVA-sensitized and PBS-challenged mice, OVA sensitization and challenge elicited airway obstruction and increased BALF eosinophils, macrophages, neutrophils, IL-4, IL-13, IL-18, and chemerin. However, OVA challenge enhanced airway obstruction and pulmonary inflammation in Cpe(fat) compared with wild-type mice. These results demonstrate that OVA sensitization and challenge enhance airway obstruction in obese mice regardless of the genetic basis of obesity, whereas the degree of OVA-induced pulmonary inflammation is dependent on the genetic modality of obesity induction. These results have important implications for animal models of asthma, as modeling the pulmonary phenotypes for subpopulations of atopic, obese asthmatics critically depends on selecting the appropriate mouse model.

Topics: Airway Obstruction; Airway Resistance; Animals; Antigens; Biomarkers; Bronchoalveolar Lavage Fluid; Carboxypeptidase H; Disease Models, Animal; Female; Genotype; Immunoglobulins; Inflammation Mediators; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Obesity; Ovalbumin; Phenotype; Pneumonia; Time Factors

(-)-Δ(9)-Tetrahydrocannabinol has been demonstrated to have beneficial effects in the airways, but its psychoactive effects preclude its therapeutic use for the treatment of airways diseases. In the present study we have investigated the effects of (-)-cannabidiol, a non-psychoactive component of cannabis for its actions on bronchial smooth muscle in vitro and in vivo. Guinea-pig bronchial smooth muscle contractions induced by exogenously applied spasmogens were measured isometrically. In addition, contractile responses of bronchial smooth muscle from ovalbumin-sensitized guinea-pigs were investigated in the absence or presence of (-)-cannabidiol. Furthermore, the effect of (-)-cannabidiol against ovalbumin-induced airway obstruction was investigated in vivo in ovalbumin-sensitized guinea-pigs. (-)-Cannabidiol did not influence the bronchial smooth muscle contraction induced by carbachol, histamine or neurokinin A. In contrast, (-)-cannabidiol inhibited anandamide- and virodhamine-induced responses of isolated bronchi. A fatty acid amide hydrolase inhibitor, phenylmethanesulfonyl fluoride reversed the inhibitory effect of (-)-cannabidiol on anandamide-induced contractions. In addition, (-)-cannabidiol inhibited the contractile response of bronchi obtained from allergic guinea-pigs induced by ovalbumin. In vivo, (-)-cannabidiol reduced ovalbumin-induced airway obstruction. In conclusion, our results suggest that cannabidiol can influence antigen-induced airway smooth muscle tone suggesting that this molecule may have beneficial effects in the treatment of obstructive airway disorders.

Topics: Airway Obstruction; Animals; Arachidonic Acids; Bronchi; Cannabidiol; Cannabinoids; Carbachol; Endocannabinoids; Female; Guinea Pigs; Histamine; In Vitro Techniques; Male; Muscle Contraction; Muscle, Smooth; Neurokinin A; Ovalbumin; Phenylmethylsulfonyl Fluoride; Polyunsaturated Alkamides

Mast cell hyperplasia has been observed in the lungs of mice with experimental asthma, but few reports have studied basophils. Here, we attempted to discriminate and quantify mast cells and basophils in the lungs in a murine asthma model, determine if both cells were increased by multiple antigen challenges and assess the roles of those cells in asthmatic responses.. Sensitized Balb/c mice were intratracheally challenged with ovalbumin four times. Mast cells and basophils in enzymatically digested lung tissue were detected by flow cytometry. An anti-FcεRI monoclonal antibody, MAR-1, was i.p. administered during the multiple challenges.. The numbers of both mast cells (IgE(+) C-kit(+) ) and basophils (IgE(+) C-kit(-) CD49b(+) ) increased in the lungs after three challenges. Treatment with MAR-1 completely abolished the increases; however, a late-phase increase in specific airway resistance (sRaw), and airway eosinophilia and neutrophilia were not affected by the treatment, although the early-phase increase in sRaw was suppressed. MAR-1 reduced antigen-induced airway IL-4 production. Basophils infiltrating the lung clearly produced IL-4 after antigen stimulation in vitro; however, histamine and murine mast cell protease 1 were not increased in the serum after the challenge, indicating that mast cell activation was not evoked.. Both mast cells and basophils infiltrated the lungs by multiple intratracheal antigen challenges in sensitized mice. Neither mast cells nor basophils were involved in late-phase airway obstruction, although early-phase obstruction was mediated by basophils. Targeting basophils in asthma therapy may be useful for an early asthmatic response.

Topics: Airway Obstruction; Airway Resistance; Animals; Antigens; Asthma; Basophils; Disease Models, Animal; Female; Interleukin-4; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Time Factors

Airway obstruction after antigen challenge is not always observed in patients with allergic asthma, even if they develop hyperresponsiveness. A similar event is observed in our guinea pig model of allergic asthma. Our aim was to study this phenomenon.. Sensitized guinea pigs were challenged with ovalbumin (OVA) 3 times every 10 days. Animals were divided into 2 groups: (1) Guinea pigs exhibiting airway obstruction after antigen challenge (R = responders), and (2) guinea pigs lacking airway obstruction response (NR = nonresponders). After the third antigen challenge, antigen-induced airway hyperresponsiveness (AI-AHR), serum OVA-specific immunoglobulins, bronchoalveolar lavage fluid (BALF) inflammatory cells, histamine, cysteinyl leukotrienes and thromboxane A2 (TxA2) BALF levels, and in vitro tracheal contraction induced by contractile mediators and OVA were evaluated.. R group consistently displayed a transient antigen-induced airway obstruction (AI-AO) as well as AI-AHR, high T×A2, histamine, OVA-IgG1, OVA-IgE and OVA-IgA levels, and intense granulocyte infiltration. NR group displayed no AI-AO and no changes in BALF measurements; nevertheless, AI-AHR and elevated OVA-IgG1 and OVA-IgA levels were observed. In all groups, histamine, TxA2 and leukotriene D4 induced a similar contraction. Tracheal OVA-induced contraction was observed only in R group. AI-AHR magnitude showed a direct association with OVA-IgG1 and OVA-IgA levels. The extent of AI-AO correlated directly with OVA-IgE and inversely with OVA-IgA levels.. Our data suggest that TxA2 and histamine participate in AI-AO likely through an IgE mechanism. AI-AHR might occur independently of AI-AO, contractile mediators release, and airway inflammatory cell infiltration, but IgA and IgG1 seem to be involved.

Topics: Airway Obstruction; Animals; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Guinea Pigs; Histamine; Humans; Immunization; Immunoglobulins; Leukotriene D4; Male; Ovalbumin; Respiratory Hypersensitivity; Thromboxane A2

Apigenin, a dietary plant-flavonoid has shown anti-inflammatory and anticancer properties, however the molecular basis of this effect remains to be elucidated. Thus we elucidated to anti-allergic effect of apigenin in ovalbumin (OVA)-induced asthma model mice. The OVA-induced mice showed allergic airway reactions. It included an increase in the number of eosinophils in bronchoalveolar lavage (BAL) fluid, an increase in inflammatory cell infiltration into the lung around blood vessels and airways, airway luminal narrowing, and the development of airway hyper-responsiveness (AHR). The administration of apigenin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, this study may provide evidence that apigenin plays a critical role in the amelioration of the pathogenetic process of asthma in mice. These findings provide new insight into the immunopharmacological role of apigenin in terms of its effects in a murine model of asthma.

Topics: Airway Obstruction; Animals; Apigenin; Asthma; Bronchoalveolar Lavage Fluid; Cell Movement; Cytokines; Enzyme-Linked Immunosorbent Assay; Eosinophil Peroxidase; Eosinophilia; Eosinophils; Female; GATA3 Transcription Factor; Immunization; Immunoglobulin E; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Ovalbumin

Humans vary markedly in their propensity to develop asthma, despite often being exposed to similar environmental stimuli. Similarly, mouse strains vary in susceptibility to airways pathology in experimental asthma. Sensitization and aerosol challenge with ovalbumin (OVA) induces eosinophil accumulation, mucus production and airways obstruction in BALB/c and C57BL/6 mice. In contrast, CBA/Ca mice show relatively little pathology. Allergen-induced production of IL-4, IL-5, IL-10 and IFN-gamma was detected in all three strains, with BALB/c mice generating the highest levels of IL-4, IL-5 and IL-10. Microarray analysis was used to identify genes differentially regulated in lung tissue after OVA challenge. Differentially regulated genes in the lungs of the asthma-susceptible C57BL/6 and BALB/c strains numbered 242 and 145, respectively, whereas only 42 genes were differentially expressed in the resistant CBA/Ca strain. In C57BL/6 mice, transcripts were enriched for adhesion molecules and this was associated with high levels of eosinophil recruitment. Differentially regulated genes in the lungs of only the asthma-susceptible strains numbered 64 and several of these have not previously been associated with asthma. Many of the genes differentially regulated in the susceptible strains were enzymes involved in inflammation. Using network analysis, mRNA for the anti-apoptotic protein survivin was found to be up-regulated in the lungs following allergen challenge. Survivin mRNA and protein were also expressed at high levels in eosinophils recovered by bronchoalveolar lavage from BALB/c and C57BL/6 mice. We propose that rapid apoptosis of lung eosinophils due to low expression of survivin contributes to the limitation of pathology in CBA/Ca mice.

Topics: Airway Obstruction; Allergens; Animals; Apoptosis; Asthma; Cell Adhesion; Cytokines; Disease Models, Animal; Eosinophils; Gene Expression Profiling; Gene Expression Regulation; Immunization; Inhibitor of Apoptosis Proteins; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Transgenic; Microarray Analysis; Microtubule-Associated Proteins; Mucus; Ovalbumin; Repressor Proteins; Species Specificity; Survivin

Helper T (Th) cells are deeply involved in the pathophysiology of bronchial asthma, such as eosinophilic inflammation, bronchial hyperresponsiveness and remodeling. However, it is still unclear how Th cells contribute to airflow limitation, another cardinal feature of bronchial asthma.. Unprimed BALB/c mice were transferred with ovalbumin (OVA)-reactive Th clones. Pulmonary function was monitored using a Buxco BioSystem Plethysmograph before and after OVA challenge.. When Th-transferred mice were challenged with OVA, enhanced pause (P(enh)), an indicator of airflow limitation was significantly increased 6 and 24 h after challenge, while no response was observed 30 min after challenge. Neither bovine serum albumin, an irrelevant antigen, challenge on Th-transferred mice nor OVA challenge on Th-non-transferred mice caused airway responses.. Th cells conferred antigen-induced airflow limitation to unprimed mice.

Topics: Airway Obstruction; Animals; Antigens; Asthma; Bronchial Hyperreactivity; Clone Cells; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Cell Antigen Receptor Specificity; T-Lymphocytes, Helper-Inducer; Time Factors

The alpha-2 adrenergic agonists clonidine and dexmedetomidine are used as an antihypertensive and a sedative, respectively. The aim of this study was to determine the effects of these agonists on ovalbumin-sensitized airway tone in guinea pigs.. The animals were divided into two groups: control and sensitized. The sensitized group received ovalbumin intraperitoneally and was boosted by exposure to aerosolized ovalbumin. The effects of the alpha-2 agonists were investigated by measuring (1) total lung resistance and (2) smooth muscle tension using a tracheal ring preparation.. In the control group, acetylcholine significantly increased total lung resistance in a dose-dependent manner. In the sensitized animals, total lung resistance was significantly higher (by 95%) at 6 mug kg-1 acetylcholine than that in the control group. Both clonidine and dexmedetomidine had a slight but significant inhibitory effect on the response curve of lung resistance at higher concentrations of carbachol, a potent muscarinic receptor agonist. Similar to the data obtained in the control group, both clonidine and dexmedetomidine significantly decreased total lung resistance and the inhibitory effects of these alpha-2 agonists on lung resistance were significantly distinguishable. Similar direct inhibitory effects of the alpha-2 agonists on carbachol-induced muscle contraction were observed in both the control and sensitized groups, the inhibitory effects in the sensitized group being significantly greater.. Both clonidine and dexmedetomidine can relax the airway even in the hyper-reactive state.

Topics: Acetylcholine; Adrenergic alpha-Agonists; Airway Obstruction; Animals; Bronchial Hyperreactivity; Carbachol; Clonidine; Dexmedetomidine; Guinea Pigs; Hypnotics and Sedatives; Lung; Male; Muscle, Smooth; Ovalbumin; Specific Pathogen-Free Organisms; Trachea

Previously, we found pulmonary gas trapping to be a rapid, simple and objective measure of methacholine-induced airway obstruction in naïve mice. In this study we extended that finding by using methacholine-induced pulmonary gas trapping to differentiate airway responses of ovalbumin-sensitized, ovalbumin-exposed (Positive Control) and ovalbumin-sensitized, sodium chloride-exposed (Negative Control) mice. Additionally, pulmonary gas trapping and enhanced pause were compared following methacholine exposure in sensitized and nonsensitized mice. Finally, we examined by nose-only inhalation the ability of the glucocorticosteroid budesonide and the peroxisome proliferator-activated receptor-gamma agonist ciglitazone to modify methacholine-induced airway responses in ovalbumin-sensitized mice. Positive Controls exhibited a 7.8-fold increase in sensitivity and a 2.4-fold enhancement in the maximal airway obstruction to methacholine versus Negative Controls. Following methacholine, individual Positive and Negative Control mouse enhanced pause values overlapped in 9 of 9 studies, whereas individual Positive and Negative Control mouse excised lung gas volume values overlapped in only 1 of 9 studies, and log[excised lung gas volume] correlated (P=0.023) with in vivo log[enhanced pause] in nonsensitized mice. Finally, budesonide (100.0 or 1000.0 microg/kg) reduced methacholine-mediated airway responses and eosinophils and neutrophils, whereas ciglitazone (1000.0 microg/kg) had no effect on methacholine-induced pulmonary gas trapping, but reduced eosinophils. In conclusion, pulmonary gas trapping is a more reproducible measure of methacholine-mediated airway responses in ovalbumin-sensitized mice than enhanced pause. Also, excised lung gas volume changes can be used to monitor drug interventions like budesonide. Finally, this study highlights the importance of running a positive comparator when examining novel treatments like ciglitazone.

Topics: Administration, Inhalation; Airway Obstruction; Animals; Anti-Asthmatic Agents; Asthma; Bronchial Hyperreactivity; Bronchodilator Agents; Budesonide; Disease Models, Animal; Dose-Response Relationship, Drug; Eosinophils; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; PPAR gamma; Thiazolidinediones

KP-496 is a novel dual antagonist for cysteinyl leukotriene receptor 1 (CysLT(1)) and thromboxane A(2) (TXA(2)) receptor (TP). The aim of this study was to evaluate the pharmacological profile of inhaled KP-496 and its effects on airway obstruction.. Antagonist activities of inhaled KP-496 were investigated using bronchoconstriction induced in guinea pigs by LTD(4) or U46619, a stable TXA(2) mimetic. Guinea pigs sensitized with injections of ovalbumin were used to assess the effects of inhaled KP-496 on bronchoconstriction induced by antigen (i.v.). Another set of guinea pigs were sensitized and challenged with ovalbumin by inhalation and the effects of inhaled KP-496 on immediate and late airway responses and airway hyperresponsiveness were investigated.. KP-496 significantly inhibited LTD(4)- and U46619-induced bronchoconstriction in a dose-dependent manner. The inhibitory effects of KP-496 (1%) were comparable to those of montelukast (a CysLT(1) antagonist, p.o., 0.3 mg kg(-1)) or seratrodast (a TP antagonist, p.o., 3 mg kg(-1)). KP-496 (1%) and oral co-administration of montelukast (10 mg kg(-1)) and seratrodast (20 mg kg(-1)) significantly inhibited antigen-induced bronchoconstriction, whereas administration of montelukast or seratrodast separately did not inhibit antigen-induced bronchoconstriction. KP-496 exhibited dose-dependent and significant inhibitory effects on the immediate and late airway responses and airway hyperresponsiveness following antigen challenge.. KP-496 exerts effects in guinea pigs which could be beneficial in asthma. These effects of KP-496 were greater than those of a CysLT(1) antagonist or a TP antagonist, in preventing antigen-induced airway obstruction.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Acetates; Administration, Inhalation; Administration, Oral; Airway Obstruction; Animals; Anti-Asthmatic Agents; Benzoates; Benzoquinones; Bronchoconstriction; Cyclopropanes; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Therapy, Combination; Guinea Pigs; Heptanoic Acids; Leukotriene Antagonists; Leukotriene D4; Lung; Male; Membrane Proteins; Ovalbumin; Prostaglandin Antagonists; Quinolines; Receptors, Leukotriene; Receptors, Thromboxane A2, Prostaglandin H2; Respiratory Hypersensitivity; Sulfides; Thiazoles; Time Factors

Interleukin 13 (IL-13) is considered to be a key driver of the development of airway allergic inflammation and remodeling leading to airway hyperresponsiveness (AHR). How precisely IL-13 leads to the development of airway inflammation, AHR, and mucus production is not fully understood. In order to identify key mediators downstream of IL-13, we administered adenovirus IL-13 to specifically induce IL-13-dependent inflammation in the lungs of mice. This approach was shown to induce cardinal features of lung disease, specifically airway inflammation, elevated cytokines, AHR, and mucus secretion. Notably, the model is resistant to corticosteroid treatment and is characterized by marked neutrophilia, two hallmarks of more severe forms of asthma. To identify IL-13-dependent mediators, we performed a limited-scale two-dimensional SDS-PAGE proteomic analysis and identified proteins significantly modulated in this model. Intriguingly, several identified proteins were unique to this model, whereas others correlated with those modulated in a mouse ovalbumin-induced pulmonary inflammation model. We corroborated this approach by illustrating that proteomic analysis can identify known pathways/mediators downstream of IL-13. Thus, we have characterized a murine adenovirus IL-13 lung model that recapitulates specific disease traits observed in human asthma, and have exploited this model to identify effectors downstream of IL-13. Collectively, these findings will enable a broader appreciation of IL-13 and its impact on disease pathways in the lung.

Topics: Adenoviridae; Adenoviridae Infections; Airway Obstruction; Animals; Cell Culture Techniques; Cell Division; Disease Models, Animal; Interleukin-13; Male; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Respiratory Function Tests; Respiratory Mucosa; Reverse Transcriptase Polymerase Chain Reaction

The Forkhead Box f1 (Foxf1) transcriptional factor (previously known as HFH-8 or Freac-1) is expressed in endothelial and smooth muscle cells in the embryonic and adult lung. To assess effects of Foxf1 during lung injury, we used CCl(4) and butylated hydroxytoluene (BHT) injury models. Foxf1(+/-) mice developed severe airway obstruction and bronchial edema, associated with increased numbers of pulmonary mast cells and increased mast cell degranulation after injury. Pulmonary inflammation in Foxf1(+/-) mice was associated with diminished expression of Foxf1, increased mast cell tryptase, and increased expression of CXCL12, the latter being essential for mast cell migration and chemotaxis. After ovalbumin (OVA) sensitization and OVA challenge, increased lung inflammation, airway hyperresponsiveness to methacholine, and elevated expression of CXCL12 were observed in Foxf1(+/-) mice. During lung development, Foxf1(+/-) embryos displayed a marked increase in pulmonary mast cells before birth, and this was associated with increased CXCL12 levels in the lung. Expression of a doxycycline-inducible Foxf1 dominant-negative transgene in primary cultures of lung endothelial cells increased CXCL12 expression in vitro. Foxf1 haploinsufficiency caused pulmonary mastocytosis and enhanced pulmonary inflammation after chemically induced or allergen-mediated lung injury, indicating an important role for Foxf1 in the pathogenesis of pulmonary inflammatory responses.

Topics: Airway Obstruction; Animals; Butylated Hydroxytoluene; Carbon Tetrachloride; Cell Degranulation; Cells, Cultured; Chemokine CXCL12; Chemotaxis; Endothelial Cells; Endothelium, Vascular; Forkhead Transcription Factors; Lung; Mast Cells; Mastocytosis; Mice; Mice, Transgenic; Ovalbumin; Pneumonia; Tryptases

This study evaluated the bronchodilating activity of the beta(2)-agonist carmoterol and the muscarinic M(3)-antagonist tiotropium, given intratracheally alone or in combination in anaesthetized artificially ventilated normal and actively sensitized guinea-pigs. Carmoterol (0.3-100pmol) and tiotropium (10-1000pmol) were superfused (0.01ml/min) for 5min before challenges with acetylcholine (20mug/kg i.v.), histamine (10mug/kg i.v.) or ovalbumin (5mg/kg i.v.). Both compounds given alone were markedly active against all the challenges. Tiotropium resulted more effective towards cholinergic challenge and carmoterol was very potent against histamine and ovalbumin-induced reaction, being effective already at 1pmol. In the presence of tiotropium, the bronchodilating activity of carmoterol was significantly augmented. The ED(50) value of carmoterol on the acetylcholine challenge was reduced by about 10 and 28 times (0.1 and 0.3pmol of tiotropium), that on the histamine one by 4.5 and 13 times (1 and 3pmol of tiotropium) and that on the ovalbumin-induced one by 8 and 25 times (10 and 30pmol of tiotropium). A positive interaction was also evident when other parameters were evaluated. The histamine-induced release of thromboxane B(2) was markedly reduced (56%, P<0.001) by combining completely ineffective doses of the two drugs (0.3 and 3pmol for carmoterol and tiotropium, respectively). In ovalbumin-challenged animals the time to death, amounting in control animals to 7.2+/-0.9min, was dose-dependently prolonged up to achieve complete protection from death with combination of 1 and 30pmol of carmoterol and tiotropium, respectively. The favorable interaction between carmoterol and tiotropium can represent a good option in the control of bronchopulmonary diseases marked by an increase of airway resistances.

Topics: Acetylcholine; Adrenergic beta-2 Receptor Agonists; Airway Obstruction; Airway Resistance; Amphetamines; Animals; Bronchoconstriction; Bronchodilator Agents; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Therapy, Combination; Guinea Pigs; Histamine; Hydroxyquinolines; Injections, Intraperitoneal; Injections, Intravenous; Injections, Subcutaneous; Male; Ovalbumin; Quinolones; Scopolamine Derivatives; Survival Analysis; Thromboxane B2; Tiotropium Bromide; Treatment Outcome

Inhaled adenosine induces airway obstruction in asthmatic but not healthy subjects, a phenomenon that is also observed in various animal species when they are immunised to a relevant antigen, but which does not occur in naïve animals. The purpose of this study was to investigate the mechanisms of airway responsiveness to adenosine receptor agonists in anaesthetised allergic guinea pigs. Inhaled adenosine 5'-monophosphate (AMP), the A1-selective adenosine receptor agonist N6-cyclopentyladenosine (CPA) and ovalbumin all caused airway obstruction in allergic guinea pigs, but not naïve animals, as assessed by changes in total lung resistance. In contrast, the A(2a)-selective (CGS 21680; 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxoamido adenosine) and A3-selective (IB-MECA; 1-deoxy-1-[6-[[3-iodophenyl)-methyl]amino]-9H-purin-9-yl]-N-methyl-beta-D-ribofuranuronamide) adenosine receptor agonists failed to elicit airway obstruction in passively sensitised guinea pigs. Airway obstruction induced by AMP or CPA was not inhibited by the H1 receptor antagonist, mepyramine (1 mg kg(-1)) in passively sensitised guinea-pigs. In contrast, airway obstruction to ovalbumin was significantly inhibited by this antagonist. Airway obstruction induced by AMP and CPA was significantly inhibited in sensitised animals chronically treated with capsaicin. In contrast, airway obstruction to ovalbumin was not inhibited by this treatment. Airway obstruction induced by AMP, CPA and ovalbumin was significantly inhibited following bilateral vagotomy or pharmacological treatment with atropine (2 mg kg(-1)). Airway obstruction to CPA was inhibited by the adenosine A1 receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX: 0.1-1 mg kg(-1)). In contrast, airway obstruction to ovalbumin was not inhibited by this treatment. These observations provide evidence indicating that AMP and CPA may induce airway obstruction in sensitised guinea pigs by a mechanism unrelated to histamine release from mast cells, but is mediated via an adenosine A1-receptor-dependent mechanism. The inhibition of AMP- and CPA-induced airway obstruction by atropine, capsaicin and bilateral vagotomy suggests a neuronal-dependent mechanism with the particular involvement of capsaicin-sensitive nerves.

Topics: Adenosine; Adenosine A1 Receptor Agonists; Adenosine A1 Receptor Antagonists; Adenosine A2 Receptor Agonists; Adenosine A2 Receptor Antagonists; Airway Obstruction; Airway Resistance; Anesthesia; Animals; Bronchoalveolar Lavage Fluid; Capsaicin; Cyclic AMP; Guinea Pigs; Histamine H1 Antagonists; Lung; Male; Neural Pathways; Ovalbumin; Pyrilamine; Respiratory Function Tests; Respiratory Hypersensitivity; Vagotomy; Xanthines

The pathophysiological characteristics of bronchial asthma consist of chronic inflammation of airways, airway hyperresponsiveness, and bronchoconstriction. Studies have shown that T helper type 2 (Th2) cytokines produced by both T cells and mast cells in the airway contribute substantially to the initiation of inflammation in both experimental and human bronchial asthma. GATA-3 is a transcription factor essential to the production of Th2 cytokines by T lymphocytes. To clarify the role of GATA-3-expressing T cells in the pathophysiology of bronchial asthma, we utilized transgenic (Tg) mice carrying the GATA-3 gene and the ovalbumin (OVA)-specific T cell receptor gene (GATA-3-Tg/OVA-Tg). Mice were intranasally administrated OVA without systemic immunization. Airway responses were analyzed with noninvasive and invasive whole body plethysmographs. GATA-3-Tg/OVA-Tg mice exhibited significantly higher IL-13 and IL-4 protein expression in the airway. Although there were no differences in the types of infiltrating cells between GATA-3-Tg/OVA-Tg and GATA-3-non-Tg/OVA-Tg mice and no significant increase in IgE level in either group compared with nontreated mice, the response after ACh inhalation was significantly elevated in GATA-3-Tg/OVA-Tg on the seventh day of intranasal treatment with OVA. This hyperresponsiveness was inhibited by 5-lipoxygenase inhibitor and IL-13 neutralization, suggesting that airway responses were induced through IL-13 and leukotriene pathway. In conclusion, airway hyperresponsiveness, a characteristic of bronchial asthma, is regulated at the level of GATA-3 transcription by T lymphocytes in vivo.

Topics: Airway Obstruction; Animals; Arachidonate 5-Lipoxygenase; Bronchoalveolar Lavage Fluid; Disease Models, Animal; DNA Primers; GATA3 Transcription Factor; Lymphocyte Activation; Mice; Mice, Transgenic; Ovalbumin; Receptors, Antigen, T-Cell; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Th2 Cells

To investigate mechanisms underlying the late asthmatic response, we developed a murine model using repetitive intratracheal antigen challenge. BALB/c mice sensitized by i.p. injection with ovalbumin+alum were challenged with ovalbumin intratracheally 4 times. The 1st challenge induced early airway obstruction peaking at 30 min but without a late response; however, the 4th challenge caused not only early but also late airway obstruction at 2-8 h. Eosinophils, and CD4+ and CD8+ T lymphocytes were increased in the airway before the 4th but not before the 1st-3rd challenges. The numbers of IgE+/CD117+ (mast) cells were also increased in the lung before the 4th challenge. Levels of Th2 cytokines were also increased in the airway. Daily administration of dexamethasone during the challenge period suppressed all these inflammatory events. Thus, this experimental late asthmatic response is associated with Th2 cytokine production from inflammatory cells recruited as a consequence of the 1st-3rd challenges.

Topics: Airway Obstruction; Airway Resistance; Alum Compounds; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokines, CC; Cytokines; Dexamethasone; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Immunoglobulin E; Immunoglobulin G; Inflammation; Injections, Intraperitoneal; Leukocytes; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-kit; Time Factors

Disodium cromoglycate (DSCG) is known to inhibit both immediate and late asthmatic responses (IAR and LAR). However, its effect on mucus hypersecretion is unknown. Using a murine model of asthma, we aimed to determine whether mucus secretion increased during IAR and LAR. We also studied the potency of DSCG in inhibiting mucus secretion and on airway eosinophilia.. Mice were subjected to initial intraperitoneal sensitization and airway challenge to ovalbumin (OVA) and then provoked by additional exposure to OVA. Some mice were pretreated with aerosolized DSCG (20 mg/ml) 1 h before the provocation with OVA. After serial measurements of enhanced pause (Penh), an indicator of airflow obstruction, serum samples and bronchoalveolar lavage fluids (BALF) were collected. Then, the lungs were excised and a morphometric analysis for mucus hypersecretion was performed.. A biphasic increase in Penh (IAR and LAR) was observed in sensitized animals after provocation with OVA. Airway eosinophilia was observed during both responses. Intraluminal mucus significantly increased during LAR, but not during IAR. DSCG significantly attenuated both IAR and LAR, and significantly inhibited the increase in intraluminal mucus during LAR, but had no effect on eosinophilia in BALF.. Our results suggest that airway hypersecretion may be involved as a component of airflow obstruction during LAR, and that this is unlikely during IAR. DSCG may be effective in reducing excessive airway mucus caused by exposure to allergens.

Topics: Airway Obstruction; Animals; Anti-Asthmatic Agents; Antigens; Asthma; Bronchoalveolar Lavage Fluid; Cromolyn Sodium; Disease Models, Animal; Eosinophilia; Eosinophils; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin

Airway hyperresponsiveness, airway eosinophilia and increased IgE levels in serum are the important characteristic features of asthma. We evaluated the potential of para-Bromophenacyl bromide (PBPB), a known phospholipase A(2) inhibitor, on allergen-induced airway hyperresponsiveness in a mouse model. We sensitized and challenged mice with ovalbumin (OVA) to develop airway hyperresponsiveness as assessed by airway constriction and airway hyperreactivity (AHR) to methacholine (MCh) induced by allergen. The mice were orally treated with PBPB (0.1, 1 and 10 mg/kg) during or after OVA-sensitization and OVA-challenge to evaluate its protective or reversal effect on airway constriction and AHR to MCh. Determination of OVA-induced airway constriction and AHR to MCh were performed by measuring specific airway conductance (SGaw) using non-invasive dual-chamber whole body-plethysmography. We observed that PBPB (1 mg/kg) significantly reduced OVA-induced airway constriction and AHR to MCh (p<0.01). PBPB (1 mg/kg) treatment significantly inhibited PLA(2) activity in the BAL fluid. Cytokine analysis of the BAL fluid revealed that PBPB caused an increase in interferon-gamma (IFN-gamma) (p<0.02) and a decrease in interleukin-4 (IL-4) (p<0.05) and interleukin-5 (IL-5) (p<0.05) levels. The OVA-specific serum IgE levels (p<0.01) and the BAL eosinophils (p<0.001) were also reduced significantly. Thus, PBPB is capable of modulating allergen induced cytokine levels and serum IgE levels, and alleviating allergen induced airway hyperresponsiveness and eosinophils in mice. These data suggest that PBPB could be useful in the development of novel agents for the treatment of allergen induced airway hyperresponsiveness.

Topics: Acetophenones; Administration, Inhalation; Aerosols; Airway Obstruction; Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Drug Evaluation, Preclinical; Eosinophils; Immunization; Immunoglobulin E; India; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Phospholipases A

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

The reversible airway hyper-reactivity (AHR) of asthma is modeled by sensitizing and challenging mice with aerosolized ovalbumin. However, the C57BL/6 murine strain does not display the large increase in circulating IgG and IgE antibodies found in human atopy and asthma. We found that commercial ovalbumin was contaminated with lipopolysaccharide (LPS) in amounts sufficient to fully activate endothelial cells in an in vitro assay of the first step of inflammation. Desensitization of TLR4 by LPS pretreatment suppressed the inflammatory effect of ovalbumin. The presence of LPS was occult, because it does not require serum presentation and, like the LPS of Salmonella minnesota, was not suppressed by polymyxin B. Purified ovalbumin did not activate endothelial cells in vitro; however, endotoxin-free ovalbumin was far more effective than commercial material in stimulating IgE production and respiratory dysfunction in a C57BL/6 murine model of AHR. Moreover, endotoxin-free ovalbumin induced lung inflammation with alveolar enlargement and destruction in a histologic pattern that differed from the changes caused by commercial, endotoxin-contaminated ovalbumin. Reconstitution of purified ovalbumin with S. minnesota LPS decreased lung inflammation, decreased changes in lung function, and suppressed anti-ovalbumin antibody production. We conclude endotoxin contaminates ovalbumin preparations and that endotoxin co-administration with the ovalbumin antigen creates a state of tolerance in a murine model of AHR. Co-exposure to endotoxin and antigen occurs in humans through organic dusts, so murine models of AHR may reflect the clinical situation, but models based on commercial ovalbumin do not accurately reflect the effect of protein antigen alone on animal physiology.

Topics: Airway Obstruction; Animals; Asthma; Disease Models, Animal; Drug Contamination; Endothelium, Vascular; Humans; Immune Tolerance; Immunity; Immunoglobulins; Lipopolysaccharides; Mice; Ovalbumin; Pneumonia; Umbilical Veins

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

Activation of nontransmembrane protein tyrosine kinases, such as Lyn and Syk, has been shown to be the earliest detectable signaling response to Fc receptor (Fc epsilon RI) cross-linking on mast cells leading to mast cell degranulation. The present study examined the effects of piceatannol (3,4,3',5'-tetrahydroxy-trans-stilbene, 10-100 microM), a Syk-selective tyrosine kinase inhibitor, on ovalbumin-induced anaphylactic contraction of isolated guinea pig bronchi and release of histamine and peptidoleukotrienes from chopped lung preparations. Pretreatment with piceatannol slightly suppressed ovalbumin-induced peak anaphylactic bronchial contraction but markedly (P<0.05) facilitated relaxation of the anaphylactically contracted bronchi. Piceatannol did not inhibit direct histamine-, leukotriene D(4)- or KCl-induced bronchial contraction, nor revert an existing anaphylactic bronchial contraction. Piceatannol, at 30 microM and above, significantly (P<0.05) prevented ovalbumin-induced release of both histamine and peptidoleukotrienes from lung fragments. Piceatannol did not inhibit exogenous arachidonic acid-induced release of peptidoleukotrienes from lung fragments. Our data show for the first time that inhibition of Syk tyrosine kinase can attenuate anaphylactic bronchial contraction in vitro, probably via inhibition of mast cell degranulation.

Topics: Airway Obstruction; Anaphylaxis; Animals; Bronchoconstriction; Enzyme Inhibitors; Enzyme Precursors; Enzyme-Linked Immunosorbent Assay; Guinea Pigs; Histamine Release; In Vitro Techniques; Injections, Intraperitoneal; Intracellular Signaling Peptides and Proteins; Leukotrienes; Lung; Male; Ovalbumin; Protein-Tyrosine Kinases; Stilbenes; Syk Kinase

The role of allergens in asthmatic inflammation is not clearly understood. To elucidate the mechanism of cockroach allergen (CRa)-induced airway disease, we studied three groups of Hartley guinea-pigs sensitized to control, ovalbumin (OA) or CRa. Parameters measured were anaphylactic antibodies by allergy skin test (AST), PCA assay and Western blot, changes in specific airway resistance (SRaw), analysis of bronchoalveolar lavage (BAL) and contracture responses of tracheal muscle (TSM) to non-specific and specific stimuli, in vitro. Both OA and CRa animals showed a similar allergic sensitization (AST and PCA), while Western blot identified several reaginic bands in CRa group compared to a single band in OA group. SRaw illustrated that CRa induce dual-asthmatic responses (4/6) in the CRa group, whereas OA induce only an early asthmatic response (3/6) in the OA group (P<0.01). The average total leukocytes in BALF of the CRa were 27.0x10(6), mostly neutrophils and eosinophils, while those of the OA showed 3.5x10(6), mostly eosinophils, respectively (P<0.0001). TSM responses to non-specific stimuli were similar in both groups (P>0.1), while the antigen-specific TSM contractions were more brisk in the OA group than those of CRa group (P<0.001). Thus, the study indicates that both CRa and OA sensitize guinea-pigs, yet CRa induces more severe and persistent late-phase inflammation than OA. This appears to be related to an influx of neutrophils rather than anaphylactic bronchospasm.

Topics: Airway Obstruction; Allergens; Anaphylaxis; Animals; Asthma; Bronchial Spasm; Bronchoalveolar Lavage Fluid; Cockroaches; Guinea Pigs; Immunization; In Vitro Techniques; Male; Muscle Contraction; Neutrophils; Ovalbumin; Passive Cutaneous Anaphylaxis; Respiratory Muscles; Trachea

Mouse models have been established mirroring key features of human bronchial asthma including airway hyperresponsiveness (AHR). Acute airway obstruction in response to an allergen challenge, however, remains to be demonstrated in these models.. A mouse model of allergic lung inflammation was employed to analyze the development of specific (allergen-induced) and nonspecific (methacholine-induced) airway obstruction.. Mice were sensitized to ovalbumin (OVA) and challenged with OVA aerosol twice each week during four weeks. Changes in lung functions were determined by noninvasive head-out body plethysmography. The development of acute airway obstruction after OVA challenge and AHR after methacholine aerosol application were assessed by a decrease in the mid-expiratory flow rate (EF(50)).. Two airway challenges were sufficient to induce AHR (5.7 vs. 15 mg/ml methacholine). Further OVA challenges reduced the baseline EF(50) from 1.85 to 1.20 ml/s (4th week) and induced acute airway obstruction. The OVA-induced obstruction was maximal in the 4th week (EF(50) = 0.91 ml/s).. The development of acute airway obstruction in allergen-sensitized mice was demonstrated by means of head-out body plethysmography. In our model, AHR was observed before the development of airway obstruction.

Topics: Airway Obstruction; Airway Resistance; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Immunoglobulin E; Immunoglobulin G; Maximal Expiratory Flow Rate; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Plethysmography, Whole Body; Tidal Volume

The contributions of histamine, cysteinyl leukotrienes (CysLTs) and thromboxane A2 (TXA2) to the asthmatic responses and the magnitudes of blood and lung eosinophilia at acute and chronic stages of our asthmatic model were comparatively determined. Guinea pigs were alternately sensitized/challenged by inhalation with ovalbumin+Al(OH)3 and ovalbumin, once every 2 weeks. Effects of mepyramine, pranlukast (a CysLT antagonist) and seratrodast (a TXA2 antagonist) on the early (EAR) and/or the late asthmatic response (LAR) were assessed at the second and fourth antigen challenges. The second challenge caused EAR but not LAR. Although the EAR was decreased at the fourth challenge, a substantial LAR was seen. Both mepyramine and seratrodast inhibited the EAR at the second challenge by approximately 50%. However, at the fourth challenge, these drugs did not inhibit the EAR. The LAR at the fourth challenge was attenuated by pranlukast and seratrodast by 45% and 40%, respectively. Both the blood and lung eosinophilia were modestly and markedly induced 5 h after the second and fourth challenges, respectively. These results strongly suggest that repetition of antigen challenge induces quantitative alterations of chemical mediators participating in the asthmatic responses and a change of the body state under which eosinophils exhibit enhanced migratory activities.

Topics: Administration, Inhalation; Airway Obstruction; Aluminum Hydroxide; Animals; Antigens; Asthma; Benzoquinones; Chromones; Eosinophilia; Guinea Pigs; Heptanoic Acids; Histamine H1 Antagonists; Leukotriene Antagonists; Lung; Male; Ovalbumin; Prostaglandin Antagonists; Pyrilamine; Thromboxane A2

High-throughput file screening against inhibition of human lung PDE4 led to the discovery of 3-ethyl-1-(4-fluorophenyl)-6-phenyl-7-oxo-4, 5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine (11) as a novel PDE4 inhibitor. Subsequent SAR development, using an eosinophil PDE assay, led to analogues up to 50-fold more potent than 11 with IC50 values of 0.03-1.6 microM. One such compound, CP-220,629 (22) (IC50 = 0.44 microM), was efficacious in the guinea pig aerosolized antigen induced airway obstruction assay (ED50 2.0 mg/kg, po) and demonstrated a significant reduction in eosinophil (55%), neutrophil (65%), and IL-1beta (82%) responses to antigen challenge in atopic monkeys (10 mg/kg, po).

Topics: Airway Obstruction; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Bronchoalveolar Lavage Fluid; Cell Count; Cell Line; Cyclic AMP; Cytokines; Dihydropyridines; Drug Evaluation, Preclinical; Eosinophils; Guinea Pigs; Humans; In Vitro Techniques; Isoenzymes; Macaca fascicularis; Molecular Conformation; Neutrophils; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Pyrazoles; Pyrrolidinones; Rolipram; Structure-Activity Relationship

The excitatory innervation of the airway smooth muscle is primarily cholinergic in nature. However, the potential neural mechanism(s) underlying airway hyperresponsiveness, one of the hallmarks of asthma, is not fully understood. In this study, cholinergic hyperresponsive Flinders sensitive line (FSL) rats and their control counterparts, Flinders resistant line (FRL) rats, were repeatedly challenged with different doses of nebulized methacholine (0, 4, 16, 64, and 256 mg/ml) for 5 min. Airway responsiveness was assessed in spontaneously breathing, unrestrained animals by means of whole body plethysmography. Increased airway responsiveness of FSL rats was evidenced as a more pronounced increase in Penh value (enhanced pause, an index of bronchoconstriction) across different concentrations of methacholine. In subsequent experiments, FSL and FRL rats were sensitized to ovalbumin and challenged with nebulized antigen. Our results indicate that the genetically transmitted cholinergic hyperresponsiveness of the FSL rat is paralleled by an increased susceptibility to allergen-induced bronchoconstriction and inflammation of the airways. This study provides further evidence that neural factors can play an important role in determining airway responsiveness and thus may be relevant for the expression of asthma. In addition, the FSL rat may be a useful model for studies of airway hyperresponsiveness.

Topics: Airway Obstruction; Animals; Asthma; Autonomic Nervous System Diseases; Bronchoalveolar Lavage Fluid; Cell Count; Female; Lung; Male; Methacholine Chloride; Ovalbumin; Parasympathetic Nervous System; Parasympathomimetics; Rats; Rats, Sprague-Dawley; Respiratory Mechanics

Airway reactivity to bronchoconstrictor mediators changes with age. We studied the effects of maturation on airway responses provoked by allergen challenge [ovalbumin (OA) 0.05, 0.15 and 0.5 mg/kg i.v] in passively sensitized immature (190 +/- 2 g: 2 weeks old) and adult guinea pigs (566 +/- 16 g: 13 weeks old). In both groups of animals, we measured both lung resistance (RL) to monitor airflow obstruction and extravasation of Evans blue dye, to quantify airway plasma exudation. Immature guinea pigs required a larger dose of OA at the challenge to induce a significant increase in RL and extravasation of Evans blue dye compared with adult guinea pigs. In addition, immature animals responded less to the lower doses of OA than adults. Pretreatment with pyrilamine, an antihistamine, suppressed the increase in RL in immature animals, whereas the remaining component of the increase in RL was seen in adult animals. Intravenous allergen challenge causes less airflow obstruction and airway plasma exudation in immature than in adult guinea pigs. Allergen-induced airflow obstruction is mediated mainly via histamine in immature animals, whereas this may not be the case in adult animals.

Topics: Age Factors; Airway Obstruction; Airway Resistance; Allergens; Animals; Asthma; Exudates and Transudates; Guinea Pigs; Histamine; Histamine H1 Antagonists; Humans; Immunization, Passive; Male; Ovalbumin; Pyrilamine; Respiratory System

In passively sensitized guinea pigs, show infusion of an amount of ovalbumin insufficient to evoke airway obstruction induces hyperreactivity of the airways. A wide range of changed responsivity was observed for different test spasmogens, with leukotriene C4 > histamine > prostaglandin F2 alpha > bradykinin > leukotriene E4 > serotonin > acetylcholine. Injection of ovalbumin as a bolus produced pronounced airway obstruction without hyperreactivity. Airway obstruction due to vascular engorgement (dextran infusion) or edema (histamine infusion) did not result in hyperreactivity. Infusion of PAF induced pronounced airway obstruction together with hyperreactivity, but with a rank order of histamine > leukotriene C4 > serotonin > bradykinin > leukotriene E4 > acetylcholine. It can be concluded that allergic airway hyperreactivity in the guinea pig is spasmogen-selective and largely independent of airway obstruction. These observations question the presumption of non-selective hyperreactivity in allergic asthma and cast doubt upon the proposal that airway hyperreactivity is secondary to airway obstruction.

Topics: Airway Obstruction; Animals; Bronchial Hyperreactivity; Bronchial Spasm; Guinea Pigs; Histamine; Immunoglobulin E; Male; Ovalbumin

In guinea pigs passively sensitized to ovalbumin, sustained (6 days) subcutaneous infusion of (+/-)salbutamol (1 mg/kg/day) induced significant airway obstruction and heightened responsivity to airway spasmogens. Of these animals, a substantial proportion (78/235) were too responsive to injected spasmogens to permit infusion of ovalbumin or died following infusion of ovalbumin; yet there were few deaths (2/166) amongst the sensitized animals not exposed to (+/-)salbutamol. In comparison to the animals not exposed to (+/-)salbutamol, infusion of ovalbumin led to exaggerated responsivity of the airways to leukotriene C4, leukotriene E4, histamine, serotonin and acetylcholine, but not to prostaglandin F2 alpha or bradykinin. The capacity of sustained exposure to high doses of (+/-)salbutamol to induce airway hyperreactivity to allergic mediators may account for an association between asthma death and regular, excessive use of sympathomimetics.

Topics: Airway Obstruction; Albuterol; Animals; Bronchial Hyperreactivity; Guinea Pigs; Male; Ovalbumin; Respiratory Function Tests

Hartley or Dunkin-Hartley guinea pigs (n = 136) were actively sensitized to ovalbumin or Ascaris suum protein by five different regimes. Specific airway resistance (sR(AW)) was measured in conscious animals by a plethysmographic procedure before, immediately following and at various intervals (up to 96 h) after aerosolized antigen or vehicle challenge. Each sensitization and challenge regime produced an immediate allergic response with positive responses (defined as a 2-fold increase in sR(AW)) in 78-100% of animals. Recovery from the immediate response followed by late-phase responses was observed in only two out of a group of four animals. The results failed to substantiate literature reports of a high incidence of late responses in the guinea pig at 4-8, 17-24 or 72 h after allergen challenge.

Topics: Aerosols; Airway Obstruction; Animals; Antigens; Ascaris; Guinea Pigs; Male; Ovalbumin; Plethysmography; Respiratory Function Tests; Respiratory Hypersensitivity

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

We have reported the development of a guinea pig animal model for pulmonary hypersensitivity to inhaled chemicals in which respirations of animals are monitored continuously for 24 hr permitting detection of immediate- (IAR) and late-onset (LAR) airway reactions. Additionally implanted temperature transmitters allow determination of accompanying febrile responses. The current study investigated the relationship between severity of IAR and occurrence of LAR in individual animals. To quantify severity, a grading system was devised which took into account time to response, increase in breathing rate, and occurrence of airway constriction (ACR). Guinea pigs were sensitized by ip injection with 1 mg ovalbumin (OA). On Day 14, inhalation challenge with 12 mg/m3 OA resulted in severe IAR in all animals. No LAR were detected. Subsequent weekly inhalation challenges with OA resulted in less severe IARs and occasionally in absence of response. Febrile reactions were not detected. On one occasion a LAR was observed. It occurred in the animal demonstrating the most severe IARs and having the highest titer of specific homocytotropic antibody. These results are consistent with the mechanism of IAR involving release of spasmogenic mediators from mast cells as a result of antigen crosslinking of surface antibody. The mechanism of the LAR is addressed in the accompanying paper in which LARs are more consistently produced in OA-sensitized guinea pigs.

Topics: Administration, Inhalation; Aerosols; Airway Obstruction; Animals; Body Temperature; Disease Models, Animal; Guinea Pigs; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immunization; Immunoglobulin E; Immunoglobulin G; Male; Monitoring, Immunologic; Ovalbumin; Plethysmography; Respiratory Hypersensitivity

The guinea pig model of pulmonary hypersensitivity, developed in this laboratory, was used to study the relationship of fever with late-onset airway responses (LAR). The preceding paper M. H. Karol, J. A. Hillebrand, and P. S. Thorne. (1989). Toxicol. Appl. Pharmacol. 100, 234-246 described production of immediate-onset responses (IAR) to ovalbumin (OA) with infrequent production of LAR (Karol et al., 1989). In the current study, LAR was produced on each of two occasions in the same animals using a sensitization regimen consisting of ip injection with 1 mg OA followed by exposure to 63 mg/m3 OA aerosol on Day 8. On Days 15 and 29, pyrilamine maleate was administered prior to inhalation challenges to block H1 receptors and to allow exposure for 20 min to 63 mg/m3 OA without histamine shock and fatal anaphylactic reactions. Each of the seven guinea pigs demonstrated IAR; four of seven additionally underwent LAR on both occasions. Late-onset responses were maximal at 4-5 hr postchallenge. Febrile responses of approximately 1 degree C (maximum 1.6 degrees C) accompanied the LAR in six of seven cases. Histopathology performed at the height of response revealed peribronchiolar eosinophilia. Pulmonary function determination indicated airflow disturbance during exhalation. These results are consistent with LAR being an airway response which is accompanied by fever and demonstrate the applicability of the guinea pig animal model to the study of mechanisms underlying IAR and LAR hypersensitivities.

Topics: Administration, Inhalation; Aerosols; Airway Obstruction; Animals; Body Temperature; Disease Models, Animal; Fever; Guinea Pigs; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immunization; Lung; Lung Volume Measurements; Male; Ovalbumin; Respiration; Respiratory Hypersensitivity

In order to assess the usefulness of methods for noninvasive study of respiratory mechanics in intubated patients better, we studied 16 anesthetized paralyzed mechanically ventilated guinea pigs previously sensitized with ovalbumin. Respiratory system elastance and resistance were determined in control and acute antigen exposed (AAE) animals by the end-inflation occlusion method (EIOM), the single-breath method (SBM), and the interrupter technique (IT). In control group, total respiratory system resistance and elastance were constant over tidal volume range and the three methods provided similar results. In AAE group elastance was also constant throughout tidal volume and measured by SBM and IT was, respectively, 93.4 and 57.9% higher than in control group. The relationships between resistive pressure and expiratory flow became curvilinear with an upward convexity. IT underestimated both elastance and resistance measurements in AAE group. Using EIOM the determination of the homogeneous and uneven components of respiratory resistance was possible in control animals, whereas in AAE group resistance was entirely represented by its uneven component.

Topics: Administration, Inhalation; Airway Obstruction; Animals; Female; Guinea Pigs; Lung Volume Measurements; Ovalbumin; Respiration; Tidal Volume

We describe the effects of the antiallergic drug cromolyn sodium and the beta 2-selective adrenoceptor agonist albuterol against early and late phase changes in specific airways conductance (sGaw) and leukocyte infiltration into the airways after allergen challenge of nonanesthetized guinea pigs. Inhalation of ovalbumin by sensitized guinea pigs induced three phases of airways obstruction: an early asthmatic response (EAR) peaking at 2 h, a late response (LAR) peaking at 17 h, and a further late response (LLAR) being observed at 72 h. The LAR was accompanied by a 13-fold rise in neutrophils and a four-fold rise in eosinophils recovered by bronchoalveolar lavage (BAL) at 17 h. By 72 h, the BAL content of neutrophils had returned to near normal, whereas eosinophil numbers had risen to 6.7-fold above baseline. Inhalation of an aerosolized solution of cromolyn, 10 mg/ml, 15 min before challenge inhibited both the EAR and LAR and the influx of neutrophils into the airways at 17 h but had no effect on eosinophil accumulation. Inhalation of cromolyn at 6 h, i.e., after the completion of the EAR, inhibited the LAR, the LLAR, and the rise in eosinophils at 72 h but did not reduce the influx of neutrophils at 17 h. Administration of cromolyn at both 15 min before and 6 h after challenge inhibited all changes in sGaw and reduced the accumulation of neutrophils at 17 h and the influx of eosinophils at 72 h. In contrast, inhalation of albuterol, 0.1 mg/ml, 15 min before allergen provocation blocked the EAR and the rise in BAL neutrophils at 17 h but did not inhibit the LAR. Inhalation of albuterol at 6 h partially reversed the LAR but had no effect on either the LLAR or cellular changes. Given at both times, albuterol inhibited the EAR and neutrophil accumulation at 17 h and partially reversed the LAR but produced no other effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Airway Obstruction; Airway Resistance; Albuterol; Allergens; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cromolyn Sodium; Guinea Pigs; Leukocytes; Male; Ovalbumin; Time Factors

BL-5255 exhibited potent activity in several models of antigen-induced immediate hypersensitivity reactions in rats and guinea pigs. The compound was effective whether administered by oral or parenteral routes and in passively and actively sensitized animals. It appeared to be readily absorbed when given orally. Localized skin and bronchoconstriction reactions in rats were inhibited by the compound by oral doses at 0.014 and 1 mg/kg, respectively. BL-5255 was protective against both IgE- and IgG-mediated reactions in the rat and guinea pig. Its effectiveness versus the systemic anaphylaxis reaction in the guinea pig appears to be due to BL-5255's ability to inhibit both the IgE and IgG1 components of the reaction.

Topics: Administration, Oral; Airway Obstruction; Animals; Cromolyn Sodium; Dose-Response Relationship, Drug; Female; Guinea Pigs; Hypersensitivity, Immediate; Immunoglobulin E; Injections, Intravenous; Kinetics; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Pyrimidinones; Rats; Rats, Inbred Strains