ovalbumin and Bronchitis

The detailed pathogenesis of eosinophilic bronchitis (EB) remains unclear. Transglutaminase 2 (TG2) has been implicated in many respiratory diseases including asthma. Herein, we aim to assess preliminarily the relationship of TG2 with EB in the context of the development of an appropriate EB model through ovalbumin (OVA) sensitization and challenge in the C57BL/6 mouse strain. Our data lead us to propose a 50 μg dose of OVA challenge as appropriate to establish an EB model in C57BL/6 mice, whereas a challenge with a 400 μg dose of OVA significantly induced asthma. Compared to controls, TG2 is up-regulated in the airway epithelium of EB mice and EB patients. When TG2 activity was inhibited by cystamine treatment, there were no effects on airway responsiveness; in contrast, the lung pathology score and eosinophil counts in bronchoalveolar lavage fluid were significantly increased whereas the cough frequency was significantly decreased. The expression levels of interleukin (IL)-4, IL-13, IL-6, mast cell protease7 and the transient receptor potential (TRP) ankyrin 1 (TRPA1), TRP vanilloid 1 (TRPV1) were significantly decreased. These data open the possibility of an involvement of TG2 in mediating the increased cough frequency in EB through the regulation of TRPA1 and TRPV1 expression. The establishment of an EB model in C57BL/6 mice opens the way for a genetic investigation of the involvement of TG2 and other molecules in this disease using KO mice, which are often generated in the C57BL/6 genetic background.

Topics: Animals; Asthma; Bronchitis; Cystamine; Cytokines; Disease Models, Animal; Eosinophils; Gene Expression Regulation; GTP-Binding Proteins; Humans; Inflammation; Lung; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Protein Glutamine gamma Glutamyltransferase 2; Transglutaminases; TRPA1 Cation Channel

Matrine has been demonstrated to attenuate allergic airway inflammation. Elevated suppressor of cytokine signaling 3 (SOCS3) was correlated with the severity of asthma. The aim of this study was to investigate the effect of matrine on SOCS3 expression in airway inflammation. In this study, we found that matrine significantly inhibited OVA-induced AHR, inflammatory cell infiltration, goblet cell differentiation, and mucous production in a dose-dependent manner in mice. Matrine also abrogated the level of interleukin (IL)-4 and IL-13, but enhanced interferon (IFN)-γ expression, both in BALF and in lung homogenates. Furthermore, matrine impeded TNF-α-induced the expression of IL-6 and adhesion molecules in airway epithelial cells (BEAS-2B and MLE-12). Additionally, we found that matrine inhibited SOCS3 expression, both in asthmatic mice and TNF-α-stimulated epithelial cells via suppression of the NF-κB signaling pathway by using pcDNA3.1-SOCS3 plasmid, SOCS3 siRNA, or nuclear factor kappa-B (NF-κB) inhibitor PDTC.. Matrine suppresses airway inflammation by downregulating SOCS3 expression via inhibition of NF-κB signaling in airway epithelial cells and asthmatic mice.

Topics: Alkaloids; Animals; Asthma; Bronchitis; Cell Line; Dose-Response Relationship, Drug; Female; Humans; Matrines; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Quinolizines; Signal Transduction; Suppressor of Cytokine Signaling 3 Protein; Tumor Necrosis Factor-alpha

Asthma is a complex highly prevalent airway disease that is a major public health problem for which current treatment options are inadequate. Recently, farnesoid X receptor (FXR) has been shown to exert anti-inflammatory actions in various disease conditions, but there have been no reported investigations of Chenodeoxycholic acid (CDCA), a natural FXR agonist, in allergic airway inflammation. To test the CDCA effectiveness in airway inflammation, ovalbumin (OVA)-induced acute murine asthma model was established. We found that lung tissue express FXR and CDCA administration reduced the severity of the murine allergic airway disease as assessed by pathological and molecular markers associated with the disease. CDCA treatment resulted in fewer infiltrations of cells into the airspace and peribronchial areas, and decreased goblet cell hyperplasia, mucus secretion and serum IgE levels which was increased in mice with OVA-induced allergic asthma. The CDCA treatment further blocked the secretion of TH2 cytokines (IL-4, IL-5 and IL-13) and proinflammatory cytokine TNF-α indicate that the FXR and its agonists may have potential for treating allergic asthma.

Topics: Animals; Asthma; Base Sequence; Bronchitis; Chenodeoxycholic Acid; Cytokines; Disease Models, Animal; DNA Primers; Female; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

We investigated the role of air humidity and allergic sensitization on the acute airway response to inhaled formaldehyde (FA) vapor. Mice were sensitized to the immunogen ovalbumin (OVA) by three intraperitoneal injections followed by two aerosol challenges, giving rise to allergic airway inflammation. Control mice were sham sensitized by saline injections and challenged by saline aerosols. Once sensitized, the mice were housed at high (85-89%) or low (<10%) relative humidity, respectively for 48h prior to a 60-min exposure to either 0.4, 1.8 or about 5ppm FA. Before, during and after exposure, breathing parameters were monitored. These included the specific markers of nose and lung irritations as well as the expiratory flow rate, the latter being a marker of airflow limitation. The sensory irritation response in the upper airways was not affected by allergic inflammation or changes in humidity. At high relative humidity, the OVA-sensitized mice had a decreased expiratory airflow rate compared to the saline control mice after exposure to approximately 5ppm FA. This is in accordance with the observations that asthmatics are more sensitive than non-asthmatics to higher concentrations of airway irritants including FA. In the dry environment, the opposite trend was seen; here, the saline control mice had a significantly decreased expiratory airflow rate compared to OVA-sensitized mice when exposed to 1.8 and 4ppm FA. We speculate that increased mucus production in the OVA-sensitized mice has increased the "scrubber effect" in the nose, consequently protecting the conducting and lower airways.

Topics: Air Pollutants; Animals; Bronchitis; Chickens; Environmental Exposure; Formaldehyde; Humidity; Inhalation Exposure; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Ventilation

The role of high-mobility group box 1 (HMGB1) in chronic allergic asthma is currently unclear. Both airway neutrophilia and eosinophilia and increase in HMGB1 expression in the lungs in our murine model of chronic asthma. Inhibition of HMGB1 expression in lung in ovalbumin (OVA)-immunized mice decreased induced airway inflammation, mucus formation, and collagen deposition in lung tissues. Analysis of the numbers of CD4(+) T helper (Th) cells in the mediastinal lymph nodes and lungs revealed that Th17 showed greater increases than Th2 cells and Th1 cells in OVA-immunized mice; further, the numbers of Th1, Th2, and Th17 cells decreased in anti-HMGB1 antibody (Ab)-treated mice. In OVA-immunized mice, TLR-2 and TLR-4 expression, but not RAGE expression, was activated in the lungs and attenuated after anti-HMGB1 Ab treatment. The results showed that increase in HMGB1 release and expression in the lungs could be an important pathological mechanism underlying chronic allergic asthma and HMGB1 might a potential therapeutic target for chronic allergic asthma.

Topics: Airway Resistance; Animals; Antibodies, Neutralizing; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Chronic Disease; Cytokines; Disease Models, Animal; Female; HMGB1 Protein; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Rabbits; Th1 Cells; Th17 Cells; Th2 Cells; Toll-Like Receptor 2; Toll-Like Receptor 4

To investigate whether eosinophilic bronchitis without airway hyperresponsiveness will develop bronchial asthma in allergic mice.. Mice were sensitized with OVA on days 0, 7, and 14, challenged on days 21 to 23 (1(st) OVA challenge), and re-challenged on days 46 to 48 (2(nd) OVA challenge), intranasally with 10 (the EB group) and 200 (the AS group) μg OVA. Lung resistance (RL) was assessed 24 h after each challenge and on day 45 followed by analysis of leukocyte distribution in the bronchoalveolar lavage (BAL) fluid and histological examination.. Twenty-four hours after the 1(st) OVA challenge, aerosolized methacholine caused a dose-dependent increase in RL in all groups. At doses ≥1.56 mg/mL, RL in the AS group was significantly higher than that of the NS-1 group (P<0.01 or 0.05) and at doses ≥12.5 mg/mL, RL was markedly higher in the AS group than that of the EB group (P<0.01). The percentage of eosinophils in both the EB group and the AS group was markedly higher than that of the control group. Twenty-four hours after the 2(nd) OVA challenge, at doses ≤12.5 mg/mL, there was no significant difference in RL among all groups (P>0.05). At doses ≥12.5 mg/mL, RL in the AS group was significantly higher than that of the control group and EB group (P<0.01 or 0.05). The percentage of eosinophils in the AS group was noticeably higher than that of the EB group(P<0.05). Furthermore, there was apparent infiltration by inflammatory cells, predominantly eosinophils, into the sub-epithelial region of the bronchus and the bronchioles and around the vessels in the EB and AS group.. Re-challenge with low doses of ovalbumin did not increase airway reactivity and failed to induce bronchial asthma in mice with ovalbumin-induced EB.

Topics: Administration, Intranasal; Aerosols; Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia

Phospholipases A2 (PLA2) hydrolyzes phospholipids, initiating the production of inflammatory lipid mediators. We have previously shown that in rats, sPLA2 and cPLA2 play opposing roles in the pathophysiology of ovalbumin (OVA)-induced experimental allergic bronchitis (OVA-EAB), an asthma model: Upon disease induction sPLA2 expression and production of the broncho-constricting CysLTs are elevated, whereas cPLA2 expression and the broncho-dilating PGE2 production are suppressed. These were reversed upon disease amelioration by treatment with an sPLA2 inhibitor. However, studies in mice reported the involvement of both sPLA2 and cPLA2 in EAB induction.. To examine the relevance of mouse and rat models to understanding asthma pathophysiology.. OVA-EAB was induced in mice using the same methodology applied in rats. Disease and biochemical markers in mice were compared with those in rats.. As in rats, EAB in mice was associated with increased mRNA of sPLA2, specifically sPLA2gX, in the lungs, and production of the broncho-constricting eicosanoids CysLTs, PGD2 and TBX2 in bronchoalveolar lavage (BAL). In contrast, EAB in mice was associated also with elevated cPLA2 mRNA and PGE2 production. Yet, treatment with an sPLA2 inhibitor ameliorated the EAB concomitantly with reverting the expression of both cPLA2 and sPLA2, and eicosanoid production.. In both mice and rats sPLA2 is pivotal in OVA-induced EAB. Yet, amelioration of asthma markers in mouse models, and human tissues, was observed also upon cPLA2 inhibition. It is plausible that airway conditions, involving multiple cell types and organs, require the combined action of more than one, essential, PLA2s.

Topics: Animals; Arachidonate 5-Lipoxygenase; Arginase; Asthma; Blotting, Western; Bronchitis; Bronchoalveolar Lavage Fluid; Chitinases; Cysteine; Dinoprostone; Disease Models, Animal; Female; Group X Phospholipases A2; Humans; Leukotrienes; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phospholipases A2, Cytosolic; Phospholipases A2, Secretory; Prostaglandin D2; Rats; Receptors, Leukotriene; Reverse Transcriptase Polymerase Chain Reaction; T-Box Domain Proteins

Resident alveolar macrophages (rAMs) residing in the bronchoalveolar lumen of the airways play an important role in limiting excessive inflammatory responses in the respiratory tract. High phagocytic activity along with hyporesponsiveness to inflammatory insults and lack of autonomous IFN-β production are crucial assets in this regulatory function. Using a mouse model of asthma, we analyzed the fate of rAMs both during and after allergic bronchial inflammation. Although nearly indistinguishable phenotypically from naïve rAMs, postinflammation rAMs exhibited a strongly reduced basal phagocytic capacity, accompanied by a markedly increased inflammatory reactivity to Toll-like receptors TLR-3 (poly I:C), TLR-4 [lipopolysaccharide (LPS)], and TLR-7 (imiquimod). Importantly, after inflammation, rAMs exhibited a switch from an IFN-β-defective to an IFN-β-competent phenotype, thus indicating the occurrence of a new, inflammatory-released rAM population in the postallergic lung. Analysis of rAM turnover revealed a rapid disappearance of naïve rAMs after the onset of inflammation. This inflammation-induced rAM turnover is critical for the development of the hyperinflammatory rAM phenotype observed after clearance of bronchial inflammation. These data document a novel mechanism of innate imprinting in which noninfectious bronchial inflammation causes alveolar macrophages to acquire a highly modified innate reactivity. The resulting increase in secretion of inflammatory mediators on TLR stimulation implies a role for this phenomenon of innate imprinting in the increased sensitivity of postallergic lungs to inflammatory insults.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Differentiation; Cells, Cultured; Cytokines; Disease Progression; Female; Inflammation Mediators; Interferon-beta; Macrophages, Alveolar; Mice; Mice, Inbred C57BL; Ovalbumin; Phagocytosis; Phenotype; Toll-Like Receptors

Eosinophilic bronchitis (EB) is a useful tool for studying airway hyperresponsiveness (AHR), but an exact EB animal model is lacking. Our objective was to establish an EB mouse model using ovalbumin (OVA). Mice were divided into asthma, normal saline (NS) control and three model groups. Asthma mice were challenged intranasally with 200 μg OVA. Model groups were challenged with one of the three OVA doses (10, 20 or 100 μg), and NS control mice received normal saline. Changes in lung resistance (R(L)), serum OVA-specific immunoglobulin-E (IgE) and differential inflammatory cell counts in bronchoalveolar lavage fluid (BALF) were determined after exposure to increasing doses of methacholine (MCh). Lung histological sections were examined for inflammatory infiltration. R(L) in the 10-μg OVA-challenged model group was not significantly different compared with the NS group at any MCh concentration but was significantly different compared with the asthma group (P < 0.01). R(L) in the other two model groups was intermediate between the asthma and NS groups. Serum OVA-specific IgE and eosinophils in BALF were increased significantly in all model and asthma groups compared with the NS group, but no significant differences were observed between model and asthma groups. Inflammatory cells were seen around bronchioles and capillaries in model and asthma groups but not the NS group. A mouse model of EB without AHR can be established by 10 μg OVA challenge and provides a useful tool for studying AHR mechanisms.

Topics: Animals; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Histocytochemistry; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia; Respiratory Function Tests

Recent studies suggest that plasminogen activator inhibitor-1 (PAI-1), a major inhibitor of the fibrinolytic system, may promote the development of asthma. To further investigate the significance of PAI-1 in the pathogenesis of asthma and determine the possibility that PAI-1 could be a therapeutic target for asthma, this study was conducted. First, PAI-1 levels in induced sputum (IS) from asthmatic subjects and healthy controls were measured. In asthmatic subjects, IS PAI-1 levels were elevated, compared with that of healthy controls, and were significantly higher in patients with long-duration asthma compared with short-duration asthma. PAI-1 levels were also found to correlate with IS transforming growth factor-β levels. Then, acute and chronic asthma models induced by ovalbumin were established in PAI-1-deficient mice and wild-type mice that received intra-airway administrations of small interfering RNA against PAI-1 (PAI-1-siRNA). We could demonstrate that eosinophilic airway inflammation and airway hyperresponsiveness were reduced in an acute asthma model, and airway remodeling was suppressed in a chronic asthma model in both PAI-1-deficient mice and wild-type mice that received intra-airway administration of PAI-1-siRNA. These results indicate that PAI-1 is strongly involved in the pathogenesis of asthma, and intra-airway administration of PAI-1-siRNA may be able to become a new therapeutic approach for asthma.

Topics: Airway Remodeling; Animals; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Drug Evaluation, Preclinical; Female; Hepatocyte Growth Factor; Humans; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Targeted Therapy; Ovalbumin; Plasminogen Activator Inhibitor 1; Pulmonary Eosinophilia; RNA Interference; RNA, Small Interfering; Sputum

Neonatal stress induces permanent physiological changes that may influence the immune system. Early-life stress increases asthma disease severity in children. We investigated the effects of early-life stress on allergic airway inflammation using a murine model of asthma coupled to maternal separation as an early-life stress stimulus.. Maternally separated (MS) and unseparated control (CON) mice were sensitized with ovalbumin (OVA) beginning at day 31 after birth.. Challenging mice with OVA increased airway hyperresponsiveness (AHR) and the number of inflammatory cells recovered in the bronchoalveolar lavage (BAL), compared to saline-challenged mice. Challenging MS mice with OVA resulted in less total inflammatory cells, eosinophils, interferon-gamma, and interleukin-4 in BAL compared to CON mice. However, MS mice challenged with OVA exhibited AHR similar to CON mice challenged with OVA. In contrast, an enhanced stress protocol (MS+) involving removal of pups from their home cages following the removal of the dam resulted in inflammatory cell accumulation and cytokine levels in the BAL similar to CON mice and higher than MS mice.. These findings indicate that the effect of early-life psychological factors on the development of airway inflammatory diseases such as asthma is very complex and depends on the quality of the psychological stress stimulus.

Topics: Aging; Animals; Animals, Newborn; Asthma; Bronchitis; Cytokines; Disease Models, Animal; Eosinophils; Maternal Deprivation; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory System; Stress, Psychological; Time

Natural killer T (NK T) cells have been shown to play an essential role in the development of allergen-induced airway hyperresponsiveness (AHR) and/or airway inflammation in mouse models of acute asthma. Recently, NK T cells have been reported to be required for the development of AHR in a virus induced chronic asthma model. We investigated whether NK T cells were required for the development of allergen-induced AHR, airway inflammation and airway remodelling in a mouse model of chronic asthma. CD1d-/- mice that lack NK T cells were used for the experiments. In the chronic model, AHR, eosinophilic inflammation, remodelling characteristics including mucus metaplasia, subepithelial fibrosis and increased mass of the airway smooth muscle, T helper type 2 (Th2) immune response and immunoglobulin (Ig)E production were equally increased in both CD1d-/- mice and wild-type mice. However, in the acute model, AHR, eosinophilic inflammation, Th2 immune response and IgE production were significantly decreased in the CD1d-/- mice compared to wild-type. CD1d-dependent NK T cells may not be required for the development of allergen-induced AHR, eosinophilic airway inflammation and airway remodelling in chronic asthma model, although they play a role in the development of AHR and eosinophilic inflammation in acute asthma model.

Topics: Acute Disease; Airway Remodeling; Airway Resistance; Allergens; Animals; Antigens, CD1d; Asthma; Bronchial Hyperreactivity; Bronchitis; Chronic Disease; Disease Models, Animal; Female; Fibrosis; Immunoglobulin E; Male; Metaplasia; Mice; Mice, Inbred BALB C; Mice, Knockout; Muscle, Smooth; Natural Killer T-Cells; Ovalbumin; Pulmonary Eosinophilia; Th2 Cells

Sublingual immunotherapy (SLIT) is effective in the treatment of a variety of allergic diseases, including bronchial asthma and rhinitis. However, how this local therapy exerts such wide effects is unclear. In this study, we comparatively examined the effect of SLIT on antigen-induced airway inflammation in bronchoalveolar and nasal cavities in mice.. Mice were treated sublingually with ovalbumin (OVA) once a day for 2 weeks. Subsequently, they were immunized with the corresponding antigen. Following intratracheal and nasal challenge with OVA, infiltration of inflammatory cells into the bronchoalveolar and nasal cavities was investigated in these mice.. Massive infiltration of eosinophils as well as neutrophils into the bronchoalveolar cavity was induced by intratracheal OVA challenge. Eosinophils accumulated in the nasal cavity, but the number of neutrophils did not significantly change in response to nasal antigen challenge. These antigen-induced airway inflammatory responses, including the increases in the numbers of eosinophils and/or neutrophils in the bronchoalveolar and nasal cavities, were clearly suppressed by SLIT.. This animal model displaying differential inflammatory responses in the bronchoalveolar and nasal cavities may be useful to elucidate the efficacy and mechanisms of SLIT against various allergic diseases.

Topics: Administration, Sublingual; Animals; Antigens; Bronchitis; Bronchoalveolar Lavage Fluid; Desensitization, Immunologic; Eosinophils; Female; Mice; Mice, Inbred BALB C; Nasal Lavage Fluid; Neutrophils; Ovalbumin; Rhinitis; Vaccination

Statins are widely used to treat hyperlipidaemia. Their immunosuppressive effect has recently been confirmed in various immune mediated disease models. However, relatively few studies have been conducted on allergic inflammation, so the precise mechanisms of their actions against allergies have not been fully clarified. On the other hand, the role of interleukin (IL)17 in immune responses has been recently highlighted, but whether statins affect IL17 production has not been well studied. The effect of pravastatin on allergic airway inflammation in a mouse model was examined to elucidate the mechanism of action, focusing on its effect on IL17 production.. BALB/c mice were immunised with ovalbumin (OVA) and then challenged with OVA aerosol. Pravastatin was delivered by intraperitoneal injection during either sensitisation or the challenge.. When delivered during systemic sensitisation, pravastatin suppressed OVA induced proliferation and production of Th2 type cytokines such as IL5 in spleen cells ex vivo and in vitro. IL17 production was also suppressed. Furthermore, pavastatin delivered during the inhalation of OVA attenuated eosinophilic airway inflammation, OVA specific IgE production in serum and OVA induced IL17 production in the thoracic lymph node. We also found that pravastatin attenuated the antigen presenting capacity of CD11c(+) cells obtained from the OVA challenged lung.. Pravastatin suppresses the systemic sensitisation to allergen with downregulation of IL17 production. It also suppresses an ongoing immune response in the airway partly by suppressing antigen presentation in the lung. Therefore, statins could be a novel therapeutic option for treatment of asthma.

Topics: Animals; Antigens; Bronchi; Bronchitis; Bronchoalveolar Lavage Fluid; Down-Regulation; Eosinophils; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Immunity, Cellular; Immunoglobulins; Interleukin-17; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Pravastatin; Respiratory Hypersensitivity; Spleen

Allergic airway inflammation (AI) is commonly associated with enhanced exhaled nitric oxide (ENO) in both humans and mice. Since mouse models are being used to understand various mechanisms of asthma, a noninvasive, simple, and reproducible method to determine ENO in mice is required for serial nonterminal assessment that can be used independent of environmental situations in which the ambient air contains substantial amounts of NO as a contaminant. The aim of this study was to noninvasively measure ENO in individual mice and to test its utility as a marker of AI in different models of allergic AI. We modified the existing ENO measuring methods by incorporating flushing and washout steps that allowed simple but reliable measurements under highly variable ambient NO conditions (1-100 ppb). This method was used to serially follow ENO in acute and chronic models of allergic AI in mice. ENO was reproducibly measured by this modified method and was positively correlated to AI in both acute and chronic models of asthma but was not independently related to airway remodeling. Resolution of AI and other related parameters in dexamethasone-treated mice resulted in reduction of ENO, further confirming this association. Restriction of allergen challenge to pulmonary but not nasal airways was associated with a smaller increase in ENO compared with allergen challenge to both. Hence, ENO can now be reliably measured in mice independent of ambient NO levels and is a valid biomarker for AI. However, nasal and pulmonary airways are likely to be independent sources of ENO, and any results must be interpreted as such.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents; Asthma; Biomarkers; Breath Tests; Bronchi; Bronchitis; Chronic Disease; Dexamethasone; Disease Models, Animal; Male; Mice; Mice, Inbred C57BL; Nitric Oxide; Ovalbumin

Exposure to allergens or air pollutants often leads to asthma exacerbations associated with aggravation of airway inflammation. Although, repeated allergen challenge often induces chronic allergic airway inflammation (CAAI) and airway remodelling, yet, the effects of brief exposure to air pollutants such as SO(2) on development of CAAI and airway remodelling remain to be clarified.. The aim of the experiment was to investigate the effects of acute neutrophilic airway inflammation induced by brief exposure to SO(2) on development of CAAI and subepithelial fibrosis (SEF) in a murine model of asthma.. Acute airway inflammation was induced by brief exposure to 50 p.p.m. SO(2) (1 h/d, 3 days). CAAI and SEF in BALB/c mice were induced by repeated challenge with ovalbumin (OVA) for 5 or 9 weeks with or without prior exposure to SO(2). Bronchoalveolar lavage fluid (BALF) eosinophilia as index of CAAI, BALF endothelin-1 (ET-1) and TGF-beta1 levels, morphometric evaluation of fibrotic area beneath subbasement membrane and lung hydroxyproline content (Hyp) as indexes of SEF were monitored.. Exposure to SO(2) led to acute neutrophilic inflammation and epithelial sloughing with profound elevation of BALF ET-1. Repeated OVA challenge resulted in CAAI and SEF along with elevation of Hyp, increase of fibrotic area beneath subbasement membrane and elevation of BALF TGF-beta1. Preceding SO(2) exposure exaggerated BALF eosinophilia, facilitated and enhanced SEF with more significant elevation of BALF ET-1 and TGF-beta1 levels compared with OVA-challenged mice without prior exposure to SO(2). The increase of Hyp was positively correlated with elevation of BALF TGF-beta1 during CAAI (r=0.842, P<0.01).. This data demonstrated that SEF developed in parallel with severity and time course of CAAI following repeated OVA challenge. SO(2)-induced acute epithelial injury and neutrophilic inflammation could enhance CAAI and promote SEF, probably through overexpression of ET-1 and TGF-beta1.

Topics: Air Pollutants; Allergens; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Chronic Disease; Endothelin-1; Female; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Pulmonary Fibrosis; Respiratory Mucosa; Sulfur Dioxide; Transforming Growth Factor beta

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

Deep inspirations (DI) have bronchodilatory and bronchoprotective effects in healthy human subjects, but these effects appear to be absent in asthmatic lungs. We have characterized the effects of DI on lung mechanics during mechanical ventilation in healthy mice and in a murine model of acute and chronic airway inflammation.. Balb/c mice were sensitized to ovalbumin (OVA) and exposed to nebulized OVA for 1 week or 12 weeks. Control mice were challenged with PBS. Mice were randomly selected to receive DI, which were given twice during the minute before assessment of lung mechanics.. DI protected against bronchoconstriction of central airways in healthy mice and in mice with acute airway inflammation, but not when OVA-induced chronic inflammation was present. DI reduced lung resistance induced by methacholine from 3.8 +/- 0.3 to 2.8 +/- 0.1 cmH2O.s.mL-1 in healthy mice and 5.1 +/- 0.3 to 3.5 +/- 0.3 cmH2O.s.mL-1 in acute airway inflammation (both P < 0.001). In healthy mice, DI reduced the maximum decrease in lung compliance from 15.9 +/- 1.5% to 5.6 +/- 0.6% (P < 0.0001). This protective effect was even more pronounced in mice with chronic inflammation where DI attenuated maximum decrease in compliance from 44.1 +/- 6.6% to 14.3 +/- 1.3% (P < 0.001). DI largely prevented increased peripheral tissue damping (G) and tissue elastance (H) in both healthy (G and H both P < 0.0001) and chronic allergen-treated animals (G and H both P < 0.0001).. We have tested a mouse model of potential value for defining mechanisms and sites of action of DI in healthy and asthmatic human subjects. Our current results point to potent protective effects of DI on peripheral parts of chronically inflamed murine lungs and that the presence of DI may blunt airway hyperreactivity.

Topics: Acute Disease; Airway Resistance; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Bronchoconstrictor Agents; Chronic Disease; Elasticity; Immunization; Inhalation; Lung; Lung Compliance; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Mechanics; Respiratory System

Active smoking increases asthma severity and is related to diminished treatment efficacy. Animal models in which inhalation of both allergen and mainstream cigarette smoke are combined can help us to understand the complex interaction between both agents. We have recently shown that, in allergic mice, the airway inflammation can be cleared by repeated allergen challenge, resulting in the establishment of a state of inhalational tolerance.. In this study, we assessed in vivo the impact of cigarette smoke on the efficacy and time course of this form of tolerance induction. We exposed sensitized mice to concurrent mainstream cigarette smoke and allergen (Ovalbumin- OVA) and measured the airway inflammation at different time points.. We first confirmed that aerosolized OVA administered for a prolonged time period (4-8 weeks) resulted in the establishment of tolerance. Concurrent OVA and smoke exposure for 2 weeks showed that tobacco smoke enhanced the Th-2 driven airway inflammation in the acute phase. In addition, the induction of the tolerance by repeated inhalational OVA challenge was delayed significantly by the tobacco smoke, since 4 weeks of concurrent exposure resulted in a more persistent eosinophilic airway inflammation, paralleled by a more mature dendritic cell phenotype. However, smoke exposure could not prevent the establishment of tolerance after 8 weeks of antigen exposure as shown by both histopathology (disappearance of the Th-2 driven inflammation) and by in vivo functional experiments. In these tolerized mice, some of the inflammatory responses to the smoke were even attenuated.. Cigarette smoke enhances acute allergic inflammation and delays, but does not abrogate the development of tolerance due to prolonged challenge with inhaled antigen in experimental asthma.

Topics: Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Chemokines; Cytokines; Flow Cytometry; Humans; Immune Tolerance; Immunoglobulin E; Lung; Male; Mice; Mice, Inbred C57BL; Muramidase; Nicotiana; Ovalbumin; Smoke; Th2 Cells; Time Factors

Recently we have shown that sound stress enhances allergic airway inflammation in a combined murine model. In the current study we investigated mediating factors and early kinetics of stress exacerbated allergic airway inflammation. Stress significantly increased allergen induced airway inflammation as identified by leukocyte numbers in BAL fluids. Eotaxin levels from stressed mice were significantly higher 24 h after stress. No differences were found for vascular or cellular adhesion molecule expression or cytokine levels. Our data indicate that the effect of stress on allergic airway inflammation might be mediated by the chemoattractant eotaxin, while Th2 cytokines and expression of adhesion molecules seem not to be differently regulated in stressed and non-stressed mice.

Topics: Allergens; Animals; Blood Cells; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Adhesion Molecules; Cell Count; Chemokine CCL11; Chemokines, CC; Corticosterone; Eosinophils; Interleukin-4; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Stress, Physiological

TNF is thought to contribute to airway hyperreactivity (AHR) and airway inflammation in asthma. However, studies with TNF-deficient or TNF receptor-deficient mice have not produced a clear picture of the role of TNF in the AHR associated with allergic inflammation in the mouse.. We used a genetic approach to investigate the contributions of TNF to antigen-induced AHR and airway inflammation in mice on the C57BL/6 background.. We analyzed features of airway allergic inflammation, including antigen-induced AHR, in C57BL/6 wild-type and TNF(-/-) mice, using 2 different methods for sensitizing the mice to ovalbumin (OVA).. In mice sensitized to OVA administered with the adjuvant aluminum hydroxide (alum), which develop IgE-independent and mast cell-independent allergic inflammation and AHR, we found no significant differences in OVA-induced AHR in C57BL/6-TNF(-/-) versus wild-type mice. By contrast, in mice sensitized to OVA without alum, which develop allergic inflammation that is significantly mast cell-dependent, C57BL/6-TNF(-/-) mice exhibited significant reductions versus wild-type mice in OVA-induced AHR to methacholine; numbers of lymphocytes, neutrophils, and eosinophils in bronchoalveolar lavage fluid; levels of myeloperoxidase, eosinophil peroxidase, and the cytokines IL-4, IL-5, and IL-17 in lung tissue; and histologic evidence of pulmonary inflammation.. In pulmonary allergic inflammation induced in mice immunized with OVA without alum, TNF significantly contributes to several features of the response, including antigen-induced inflammation and AHR.. Our findings in mice support the hypothesis that TNF can promote the allergic inflammation and AHR associated with asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Cytokines; Disease Models, Animal; Immunoglobulin E; Lung; Mast Cells; Mice; Mice, Mutant Strains; Ovalbumin; Receptors, Tumor Necrosis Factor, Type I; Receptors, Tumor Necrosis Factor, Type II; Respiratory Hypersensitivity; Tumor Necrosis Factor-alpha

Sublingual immunotherapy is a noninvasive and efficacious treatment of type I respiratory allergies. A murine model of sublingual immunotherapy is needed to understand better the immune mechanisms involved in successful immunotherapy and to assess second-generation candidate vaccines.. Herein, we developed a therapeutic murine model of sublingual immunotherapy in which we document the value of mucoadhesive formulations to enhance treatment efficacy.. BALB/c mice were sublingually treated with soluble or formulated ovalbumin before or after sensitization with ovalbumin. Airways hyperresponsiveness and lung inflammation were assessed by whole-body plethysmography and lung histology, respectively. Humoral and cellular immune responses were monitored by ELISA and ELISPOT techniques.. Prophylactic sublingual administration of ovalbumin completely prevents airways hyperresponsiveness as well as IL-5 secretion and IgE induction. Therapeutic administration of ovalbumin as a solution via either the sublingual or oral route has a limited efficacy. In contrast, sublingual application of ovalbumin formulated with maltodextrin to enhance mucosal adhesion results in a major reduction of established airways hyperresponsiveness, lung inflammation, and IL-5 production in splenocytes. This mucoadhesive formulation significantly enhances ovalbumin-specific T-cell proliferation in cervical but not mesenteric lymph nodes, and IgA production in the lungs.. A mucoadhesive maltodextrin formulation of ovalbumin enhances priming of the local mucosal immune system and tolerance induction via the sublingual route.. Mucoadhesive formulations offer the opportunity to improve dramatically sublingual immunotherapy in human beings, most particularly by simplifying immunization schemes.

Topics: Administration, Sublingual; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Cell Proliferation; Female; Immunization; Immunoglobulin A; Immunoglobulin E; Immunotherapy; Interleukin-10; Interleukin-5; Lung; Lymph Nodes; Mice; Mice, Inbred BALB C; Mouth Mucosa; Ovalbumin; Respiratory Mucosa; Spleen; T-Lymphocytes; Tissue Adhesives; Treatment Outcome

There has been increasing evidence suggesting the involvement of angiotensin II (Ang II) and type 1 Ang II receptors (AT1) in the pathogenesis of bronchial asthma. However, whether such an involvement would promote or suppress the pathophysiology of asthma is controversial.. The aim of this study was to investigate the role of AT1 in the development of allergic airway inflammation.. Agtr1a+/+ [wild-type C57BL/6 mice (WT)] and Agtr1a-/- mice [AT1a knockout mice (AT1aKO)] with a genetic background of C57BL/6 were systemically sensitized to ovalbumin (OVA), followed by OVA inhalation. OVA-specific IgE in serum obtained just before the inhalation was measured. Bronchoalveolar lavage (BAL) fluid and lung tissues were obtained at various time-points. Cell numbers and differentiation, and cytokine contents in BAL fluids were determined. Peribronchial accumulation of eosinophils and mucus inclusions in the bronchial epithelium were evaluated in lung tissues stained histochemically. Cell numbers and differentiation in BAL fluids of the mice were also determined after lipopolysaccharide (LPS) inhalation.. The levels of OVA-specific IgE in AT1aKO were significantly higher than those in WT. The numbers of total cell, eosinophils and lymphocytes in BAL fluids 7 days after OVA inhalation in AT1aKO were significantly higher than those in WT. Airway inflammation in bronchial tissues in terms of eosinophil accumulation and mucus hypersecretion in AT1aKO was also stronger than in WT. The contents of IL-4, IL-5 and IL-13, but not IFN-gamma, in BAL fluids of AT1aKO were significantly higher than those of WT. In contrast, neutrophil accumulation in BAL fluids after LPS inhalation was significantly higher in WT than in AT1aKO.. AT1a might be involved in the negative regulation of the development of allergic airway inflammation through polarizing the T-helper (Th) balance towards Th1 predominance. Therefore, it would be of clinical importance to investigate the effects of long-term administration of AT1 blockers on the Th1/Th2 balance in hypertensive patients with bronchial asthma.

Topics: Animals; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Count; Cell Differentiation; Cytokines; Disease Models, Animal; Immunoglobulin E; Lipopolysaccharides; Lung; Lymphocytes; Macrophages, Alveolar; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Mucus; Neutrophils; Ovalbumin; Receptor, Angiotensin, Type 1

To investigate the effect of Earthworm decoction on the airway inflammation of experimental bronchial asthma in guinea pigs and inquire into the mechanism in the decoction.. Forty-eight guinea pigs were randomly divided into six groups: the control group, the model group, the dexamethasone group, the Xiaoqinglong decoction group, the earthworm decoction large dosage group and the Earthworm decoction low dosage group, 8 guinea pigs in each group. Except the control group, the other groups were sensitized with ovalbumin (OVA) by a combination of intraperitional injection and repeated intranasal challenges to establish the guinea pigs asthma model. However, in the control group, normal saline was used. The morphological changes of bronchial tube, the lung tectology and the inflammation germ cell quantity of eosinophils (Eos), lymphocytes (Ly), neutrophils (Neu) and total blood cells in the blood and bronchoalveolar lavaga fluid (BALF) were examinated in each group respectively.. The levels of Eos, Ly, Neu and total cell quantity in the blood and BALF after the earthworm decoction treatment in the large dosage group were significantly lower than those in the model group (P <0.01), and in the low dosage group were lower too (P <0.05). The Earthworm decoction large dosage could obviously improve the bronchial tube epidermis damage, the mucous membrane gland proliferation and hydrops, asthma pathology change and basilar membrane accumulation. Eos apoptosis was obsered in the bronchoalveolar, blood and BALF. The Earthworm decoction small dosage had a similar effect but slightly to the large dosage.. The Earthworm decoction can lighten the airway inflammation in asthmatic guinea pigs, its mechanism is related with the inhibition of Eos infiltration, acceleration of Eos apoptosis and improvement of the bronchial tube and the lung tectology changes. The effect of the decoction is dose-dependent.

Topics: Animals; Apoptosis; Asthma; Bronchi; Bronchitis; Bronchoalveolar Lavage Fluid; Dose-Response Relationship, Drug; Drug Combinations; Drugs, Chinese Herbal; Eosinophils; Guinea Pigs; Leukocyte Count; Materia Medica; Neutrophils; Oligochaeta; Ovalbumin; Plants, Medicinal; Random Allocation

Recent epidemiological studies have shown that growing up on a traditional farm provides protection from the development of allergic disorders such as hay fever and allergic asthma. We present experimental evidence that substances providing protection from the development of allergic diseases can be extracted from dust collected in stables of animal farms.. Stable dust was collected from 30 randomly selected farms located in rural regions of the Alps (Austria, Germany and Switzerland). The dust was homogenised with glass beads and extracted with physiological sodium chloride solution. This extract was used to modulate immune response in a well established mouse model of allergic asthma.. Treatment of mice by inhalation of stable dust extract during sensitisation to ovalbumin inhibited the development of airway hyperresponsiveness and airway eosinophilia upon challenge, as well as the production of interleukin 5 by splenocytes and of antigen specific IgG(1) and IgE. Dust extract also suppressed the generation of human dendritic cells in vitro. The biological activity of the dust extract was not exclusively mediated by lipopolysaccharide.. Stable dust from animal farms contains strong immune modulating substances. These substances can interfere with the development of both cellular and humoral immunity against allergens, thus suppressing allergen sensitisation, airway inflammation, and airway hyperresponsiveness in a murine model of allergic asthma.

Topics: Allergens; Animals; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Cytokines; Dendritic Cells; Dust; Female; Flow Cytometry; Immunization; Immunoglobulins; Inhalation Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Spleen

Eosinophils are known to be the important effector cells in asthmatic airway inflammation. The purpose of this study was to investigate the effects of nobiletin, a polymethoxyflavonoid, on eosinophilic airway inflammation of asthmatic rats, and explore its possible mechanisms. Animals were actively sensitized by subcutaneous injection of ovalbumin (OVA). The inflammation in lung tissues of asthmatic rats was observed by hematoxylin and eosin (HE) staining. The eosinophils in blood and BALF were separated by Percoll density gradient centrifugation and counted under microscope. The level of Eotaxin was detected by enzyme-linked immunosorbent assay (ELISA). In addition, the apoptosis of eosinophils was labeled by TdT-mediated dUTP nick end labeling (TUNEL) technique, the semi-quantitative detection for Fas mRNA expression of eosinophils was performed by reverse transcription-polymerase chain reaction (RT-PCR). The airway inflammation of asthmatic rats pretreated with nobiletin was obviously alleviated. Nobiletin (1.5 and 5.0 mg/kg given intraperitoneally) significantly reduced OVA-induced increases in eosinophils, remarkably lowered the level of Eotaxin in blood and broncho-alveolar lavage fluid (BALF) of asthmatic rats. On the other hand, semi-quantitative RT-PCR analysis for Fas of eosinophils from OVA aerosol-challenged sensitized rats showed that Fas mRNA expression of eosinophils was obviously enhanced by nobiletin. Meanwhile, the apoptosis index of cultured eosinophils was significantly elevated after treatment with different doses of nobiletin. These results indicated that nobiletin could inhibit the eosinophilic airway inflammation. Lowering the levels of Eotaxin, relieving airway infiltration of eosinophils and promoting apoptosis of eosinophils by enhancing expression of Fas mRNA may be important mechanisms for nobiletin to antagonize eosinophilic airway inflammation of asthmatic rats.

Topics: Animals; Anti-Asthmatic Agents; Antioxidants; Apoptosis; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Chemokines, CC; Disease Models, Animal; Dose-Response Relationship, Drug; Eosinophils; fas Receptor; Flavones; Injections, Intraperitoneal; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; RNA, Messenger

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

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

T helper (Th) cells play major roles in orchestrating asthmatic airway inflammation, but the molecular mechanisms controlling Th-cell recruitment to the airways remain incompletely defined. Innate immunity contributes importantly to the recruitment of effector T cells into sites of inflammation. To understand better the role of innate immune signals in the development of airway inflammation, we used a murine model in which lipopolysaccharide (LPS) contaminating the antigen is thought to trigger Toll-like receptor 4 (TLR4). To investigate the importance of the TLR4-signaling pathway in induction of lung inflammation, we compared recruitment of adoptively transferred ovalbumin-specific Th1 and Th2 cells in wild-type and TLR4 mutant (TLR4m) mice after intranasal or aerosol challenge. Intranasal challenge of TLR4m mice with ovalbumin resulted in decreased recruitment of Th1 and Th2 cells compared with that of wild-type mice. The numbers of Th1 and Th2 cells recruited to the airways of TLR4m mice were less profoundly reduced after aerosol ovalbumin challenge. Comparing the effects of altering the dose of ovalbumin with that of LPS suggested that both contribute to the magnitude of the response in wild-type mice. Our findings demonstrate the importance of both antigen and endotoxin acting in a synergistic manner in the development of airway inflammation.

Topics: Administration, Inhalation; Administration, Intranasal; Adoptive Transfer; Animals; Antigens; Asthma; Bronchitis; Cell Proliferation; Dose-Response Relationship, Drug; Drug Synergism; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Mice, Transgenic; Neutrophils; Ovalbumin; Signal Transduction; Th1 Cells; Th2 Cells; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Overactivation of nuclear factor kappaB (NF-kappaB) orchestrates airway eosinophilia, but does not dampen airway hyperresponsiveness in asthma. NF-kappaB repression by arsenic trioxide (As2O3) contributes to apoptosis of eosinophils (EOS) in airways. Here we provide evidence that As2O3 abrogates allergen (OVA)-induced airway eosinophilia by modulating the expression of IkappaBalpha, an NF-kappaB inhibitory protein, and decreases the airway hyperresponsiveness.. Using a murine model of asthma, the airway hyperresponsiveness was conducted by barometric whole-body plethysmography. Airway eosinophilia, OVA-specific IgE in serum, and chemokine eotaxin and RANTES (regulated upon activation, normal T cell expressed and secreted) in bronchoalveolar lavage fluid were measured by lung histology, Diff-Quick staining, and ELISA. Chemokine-induced EOS chemotactic activity was evaluated using EOS chemotaxis assay. Electrophoretic mobility shift assay and Western blot analysis were performed to assess pulmonary NF-kappaB activation and IkappaBalpha expression, respectively.. As2O3 attenuated the allergen-induced serum IgE, chemokine expression of eotaxin and RANTES, and the EOS recruitment in bronchoalveolar lavage fluid, which is associated with an increased IkappaBalpha expression as well as a decreased NF-kappaB activation. Also, As2O3 suppressed the chemotaxis of EOS dose-dependently in vitro. Additionally, As2O3 significantly ameliorated the allergen-driven airway hyperresponsiveness, the cardinal feature underlying asthma.. These findings demonstrate an essential role of NF-kappaB in airway eosinophilia, and illustrate a potential dissociation between airway inflammation and hyperresponsiveness. As2O3 likely exerts its broad anti-inflammatory effects by suppression of NF-kappaB activation through augmentation of IkappaBalpha expression in asthma.

Topics: Allergens; Animals; Arsenic Trioxide; Arsenicals; Asthma; Bronchitis; Female; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Oxides; Pulmonary Eosinophilia; Respiratory Hypersensitivity

T(H)2-mediated allergic asthma is characterized by eosinophilia, mucus overproduction, and airway hyperresponsiveness (AHR). Although it is clear that T(H)2 cells and their cytokines play an important role in AHR, the roles of T(H)1 cells and neutrophils in AHR are controversial.. We sought to determine the roles of T(H)1 cells and neutrophils in AHR.. Ovalbumin-specific CD4(+) T cells were purified from DO11.10 mice, differentiated into T(H)1 cells, and injected into naive BALB/c, IL-4RalphaKO, or IL-8RKO mice. After ovalbumin antigen challenge, cytokine mRNA levels in lung samples, as well as inflammatory cell types and numbers in bronchoalveolar lavage fluid (BALF), were determined. AHR was assessed by measuring resistance in tracheostomized mice and enhanced pause in freely moving mice.. T(H)1 cells induced AHR as robust as T(H)2 cells. They also induced lung inflammation dominated by neutrophils. Neither AHR nor inflammation were reduced when T(H)1 cells were transferred into IL-4RalphaKO mice. When IL-8RKO mice were used as recipients of T(H)1 cells, neutrophilia was greatly reduced, but the AHR was as strong as that seen in wild-type mice. On the other hand, dexamethasone treatment had no effect on neutrophilia but has significantly reduced AHR. Reduction in AHR was accompanied by a reduction in the numbers of lymphocytes and macrophages in BALF.. T(H)1 cells can induce strong AHR independent of IL-4 and IL-13. The AHR is associated with the presence of lymphocytes and macrophages, but not neutrophils, in BALF. Our results point to a pathway whereby T(H)1 cells mediate AHR independent of neutrophilic inflammation.

Topics: Adoptive Transfer; Animals; Bronchial Hyperreactivity; Bronchitis; Cytokines; Dexamethasone; Glucocorticoids; Interleukin-13; Mice; Mice, Inbred BALB C; Mice, Knockout; Mucus; Neutrophils; Ovalbumin; Receptors, Interleukin-8A; RNA, Messenger; Severity of Illness Index; Th1 Cells; Th2 Cells

We examined the reversibility of several changes in the lungs and airways of mice immediately after exposure to ovalbumin aerosol and after a period of recovery breathing clean air.. Mice were exposed for 1, 2, 4, 6, 8, or 10 weeks, with recovery in clean air for 1-3 weeks.. Airway collagen content, exhaled NO, airway mucous cell hyperplasia, and lung lavage inflammatory cell content increased upon exposure to ovalbumin aerosol. All parameters except airway fibrosis decreased partially or completely to control values with recovery in clean air.. Airway mucous cell hypertrophy and hyperplasia appear to be completely reversible after recovery in clean air, while exhaled NO and airway inflammation appear to be mostly reversible, except for persistence of lymphocytes in the lung lavage fluid. Airway fibrosis appears to be reversible when mice are exposed to ovalbumin aerosol for periods of up to 4 weeks of exposure, but becomes irreversible after 6 or more weeks of exposure.

Topics: Administration, Inhalation; Animals; Bronchial Diseases; Bronchitis; Collagen; Drug Administration Schedule; Exhalation; Female; Fibrosis; Hyperplasia; Hypertrophy; Male; Mice; Mice, Inbred BALB C; Nitric Oxide; Ovalbumin; Pneumonia; Pulmonary Fibrosis; Respiratory Mucosa

Leukotriene B4 (LTB4) is a rapidly synthesized, early leukocyte chemoattractant that signals via its cell surface receptor, leukotriene B4 receptor 1 (BLT1), to attract and activate leukocytes during inflammation. A role for the LTB4-BLT1 pathway in allergen-induced airway hyperresponsiveness and inflammation is not well defined.. To define the role of the LTB4 receptor (BLT1) in the development of airway inflammation and altered airway function.. BLT1-deficient (BLT1 -/-) mice and wild-type mice were sensitized to ovalbumin by intraperitoneal injection and then challenged with ovalbumin via the airways. Airway responsiveness to inhaled methacholine, bronchoalveolar lavage fluid cell composition and cytokine levels, and lung inflammation and goblet cell hyperplasia were assessed.. Compared with wild-type mice, BLT1 -/- mice developed significantly lower airway responsiveness to inhaled methacholine, lower goblet cell hyperplasia in the airways, and decreased interleukin (IL)-13 production both in vivo, in the bronchoalveolar lavage fluid, and in vitro, after antigen stimulation of lung cells in culture. Intracellular cytokine staining of lung cells revealed that bronchoalveolar lavage IL-13 levels and numbers of IL-13(+)/CD4+ and IL-13(+)/CD8+ T cells were also reduced in BLT1 -/- mice. Reconstitution of sensitized and challenged BLT1 -/- mice with allergen-sensitized BLT1 +/+ T cells fully restored the development of airway hyperresponsiveness. In contrast, transfer of naive T cells failed to do so.. These data suggest that BLT1 expression on primed T cells is required for the full development of airway hyperresponsiveness, which appears to be associated with IL-13 production in these cells.

Topics: Allergens; Animals; Antibody Formation; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; CD3 Complex; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cytokines; Female; Interleukin-13; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Receptors, Leukotriene B4; T-Lymphocytes

The pathophysiology of asthma involves the action of inflammatory/allergic lipid mediators formed following membrane phospholipid hydrolysis by phospholipase A(2) (PLA(2)). Cysteinyl leukotrienes are considered potent inducers of bronchoconstriction and airway remodelling. Ovalbumin (OVA) induced bronchoconstriction in rats is associated with increased secretory PLA(2) (sPLA(2)) activation and cysteinyl leukotriene production, together with suppression of cytosolic PLA(2) and prostaglandin E(2). These processes are reversed when the animals are pretreated systemically with an extracellular cell impermeable sPLA(2) inhibitor which also suppresses the early allergic reaction to OVA challenge. In this study we examine the capacity of the sPLA(2) inhibitor to ameliorate inflammatory and allergic manifestations (early and late bronchoconstriction) of OVA induced allergic bronchitis in rats when the inhibitor was administered by inhalation to confine it to the airways.. Rats sensitised with OVA were treated with the sPLA(2) inhibitor hyaluronic acid-linked phosphatidyl ethanolamine (HyPE). The rats were divided into four groups (n = 10 per group): (1) naïve controls (no sensitisation/no treatment); (2) positive controls (sensitisation + challenge with OVA inhalation and subcutaneous injection of 1 ml saline before each challenge; (3) sensitisation + challenge with OVA and HyPE inhalation before every challenge; and (4) sensitisation + challenge with OVA and treatment with subcutaneous dexamethasone (300 mug) before each challenge as a conventional reference. Another group received no treatment with HyPE during the sensitisation process but only before or after challenge of already sensitised rats. Pulmonary function was assessed and changes in the histology of the airways, levels of cysteinyl leukotrienes in BAL fluid, and the production of nitric oxide (No) and tumour necrosis factor alpha (TNFalpha) by BAL macrophages were determined.. Inhalation of HyPE markedly suppressed OVA induced early and late asthmatic reactions as expressed by bronchoconstriction, airway remodelling (histology), cysteinyl leukotriene level in BAL fluid, and production of TNFalpha and NO by BAL macrophages. OVA induced bronchoconstriction in sensitised non-pretreated rats was also inhibited by inhalation of HyPE either before or after the challenge.. These findings confirm the pivotal role of sPLA(2) in the pathophysiology of both the immediate allergic response and the inflammatory asthmatic process. Control of airway sPLA(2) may be a new therapeutic approach to the treatment of asthma.

Topics: Administration, Inhalation; Animals; Biomarkers; Blood Proteins; Bronchitis; Ovalbumin; Rats; Respiratory Hypersensitivity

Exposure to diesel exhaust particles (DEP) during the sensitization process has been shown to increase antigen-specific IgE production and aggravate allergic airway inflammation in human and animal models. In this study, we evaluated the effect of short-term DEP exposure on ovalbumin (OVA)-mediated responses using a post-sensitization model. Brown Norway rats were first exposed to filtered air or DEP (20.6 +/- 2.7 mg/m3) for 4 h/day for five consecutive days. One day after the final air or DEP exposure (day 1), rats were sensitized with aerosolized OVA (40.5 +/- 6.3 mg/m3), and then again on days 8 and 15, challenged with OVA on day 29, and sacrificed on days 9 or 30, 24 h after the second OVA exposure or the final OVA challenge, respectively. Control animals received aerosolized saline instead of OVA. DEP were shown to elicit an adjuvant effect on the production of antigen-specific IgE and IgG on day 30. At both time points, no significant airway inflammatory responses and lung injury were found for DEP exposure alone. However, the OVA-induced inflammatory cell infiltration, acellular lactate dehydrogenase activity and albumin content in bronchoalveolar lavage (BAL) fluid, and numbers of T cells and their CD4+ and CD8+ subsets in lung-draining lymph nodes were markedly reduced by DEP on day 30 compared with the air-plus-OVA exposure group. The OVA-induced nitric oxide (NO) in the BAL fluid and production of NO, interleukin (IL)-10, and IL-12 by alveolar macrophages (AM) were also significantly lowered by DEP on day 30 as well as day 9. DEP or OVA alone decreased intracellular glutathione (GSH) in AM and lymphocytes on days 9 and 30. The combined DEP and OVA exposure resulted in further depletion of GSH in both cell types. These results show that short-term DEP exposure prior to sensitization had a delayed effect on enhancement of the sensitization in terms of allergen-specific IgE and IgG production, but caused an attenuation of the allergen-induced airway inflammatory responses.

Topics: Adjuvants, Immunologic; Air Pollutants; Allergens; Animals; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Glutathione; Immunoglobulin E; Inhalation Exposure; L-Lactate Dehydrogenase; Lymph Nodes; Macrophages, Alveolar; Male; Nitric Oxide; Ovalbumin; Rats; Rats, Inbred BN; T-Lymphocyte Subsets; Vehicle Emissions

To determine the expression of nerve growth factor (NGF), tyrosine kinase receptor A (trkA), and pan-neurotrophin receptor (p75) in the lung tissues in asthmatic rats, and to explore their effects on the airway inflammation.. Thirty-two SD rats were randomly divided into 4 groups: the control, asthma, NGF and anti-NGF groups. The asthmatic model was established by the inhalation and injection of ovalbumin. The total cell count and differential cell count in the bronchoalveolar lavage fluid (BALF) were performed. The pathologic changes in the lung tissues of the 4 groups was detected by HE staining. The NGF mRNA expression in the lung tissues of the asthma and control groups was determined by reverse transcription-polymerase chain reaction (RT-PCR). The changes of trkA and p75 mRNA expressions in the lung tissues in the 4 groups were also investigated by RT-PCR.. Compared with the control group, the BALF total cell, the BALF eosinophils (Eos), and the BALF lymphocytes (Lyms) significantly increased (All P <0. 001) in the asthma group; and the lung tissues of the asthma group had more infiltrating inflammatory cells. Not only the expression of NGF mRNA, but also its receptors trkA and p75 mRNA in the lung tissues were significantly higher in the asthma group than those in the control group (All P < 0.01). Positive correlation was found between the expression of NGF mRNA and the BALF total cell, the BALF Lyms in the asthma group. Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the NGF group significantly increased (All P < 0.01), and the lungs of the NGF group had apparent inflammatory changes. The expre-ssions of p75 and trkA mRNA were enhanced significantly (All P < 0.05). Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the anti-NGF group significantly decreased (All P < 0.001), and the lungs of the anti-NGF group showed alleviative inflammatory changes. The expre-ssions of p75 and trkA mRNA significantly decreased (All P < 0.01).. In lungs of asthmatic rats, the elevated expression of NGF mRNA is closely related to the airway inflammation. NGF can upregulate the expressions of p75 and trkA mRNA in asthmatic rats, and then may promote their role in the airway neuronal inflammation in asthma.

Topics: Animals; Asthma; Bronchitis; Lung; Male; Nerve Growth Factor; Ovalbumin; Random Allocation; Rats; Rats, Sprague-Dawley; Receptor, trkA; Receptors, Nerve Growth Factor; RNA, Messenger

MX-68 is a newly synthesized antifolate, which is a derivative of methotrexate (MTX). In this paper, the effect of MX-68 on allergic airway responses in mice and guinea-pigs was studied. In the first experiment, antigen-induced airway inflammation and airway hyperresponsiveness (AHR) to acetylcholine in mice were examined and compared with the effects of classical antifolate methotrexate and prednisolone. Mice were sensitized with ovalbumin as an antigen and challenged with ovalbumin inhalation three times. After the last inhalation, AHR and airway inflammation were observed. An increase in Th2 cytokines (IL-4 and IL-5) and a decrease in a Th1 cytokine (IFN-gamma) in the bronchoalveolar lavage fluid (BALF), as well as an elevation of the immunoglobulin level in serum, were observed in sensitized mice. Oral administration of MX-68 had no effect on changes of body weight, but prednisolone reduced body weight during the experiment. The antigen-induced AHR and increases in the number of eosinophils and lymphocytes in BALF were significantly inhibited by MX-68. MX-68 interfered with the elevation of IL-4 and IL-5 levels in BALF, but had no effect on the decrease in IFN-gamma. Moreover, MX-68 significantly inhibited the elevation of serum IgE and IgG levels. In the guinea-pig model for bronchial asthma, biphasic increases in airway resistance (immediate asthmatic response, IAR, and late asthmatic response, LAR), as well as accumulated inflammatory cells in BALF, were observed after repeated antigen challenge. These asthmatic responses and inflammatory signs were significantly decreased by administration of MX-68. These results suggest that MX-68 obviously has an anti-inflammatory effect in an animal model of asthma and would be useful clinically for the treatment of bronchial asthma.

Topics: 2-Aminoadipic Acid; Acetylcholine; Administration, Inhalation; Allergens; Animals; Body Weight; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Dogs; Dose-Response Relationship, Drug; Guinea Pigs; Immunoglobulins; Injections, Intraperitoneal; Interferon-gamma; Interleukin-4; Interleukin-5; Male; Methotrexate; Mice; Mice, Inbred BALB C; Ovalbumin; Prednisolone

Aerosolized monoclonal antibodies (mAbs) specific for T-cell receptors (TCR) were used to manipulate T-cell function in airways of ovalbumin (OVA)-sensitized and -challenged mice with airway hyperresponsiveness (AHR). The inhaled mAbs were found to be effective at low doses, had little or no systemic effect and specifically abrogated both effector and regulatory functions of the targeted T cells. Specific mAbs targeting alphabeta T cells suppressed and those targeting gammadelta T cells enhanced AHR. Moreover, specific mAbs directed against subsets of gammadelta T cells varied in their effect on AHR. Using this approach of targeting either alphabeta or gammadelta T cells reduced airway eosinophila, although the effect of mAbs specific for alphabeta T cells was stronger. The use of aerosolized anti-TCR mAbs may offer an effective approach for the treatment of airway inflammation and AHR.

Topics: Administration, Inhalation; Airway Resistance; Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody Specificity; Bronchial Hyperreactivity; Bronchitis; Disease Models, Animal; Dose-Response Relationship, Drug; Flow Cytometry; Lung; Lymphocyte Count; Lymphocyte Depletion; Methacholine Chloride; Mice; Mice, Inbred C57BL; Ovalbumin; Receptors, Antigen, T-Cell; Receptors, Antigen, T-Cell, gamma-delta; Respiratory Hypersensitivity; T-Lymphocyte Subsets

Allergic asthma is a complex syndrome characterized by airway obstruction, airway inflammation and airway hyper-responsiveness (AHR). Using a mouse model of allergen-induced AHR, we previously demonstrated that CD8-deficient mice develop significantly lower AHR, eosinophilic inflammation and interleukin (IL)-13 levels in bronchoalveolar lavage fluid compared with wild-type mice. These responses were restored by adoptive transfer of antigen-primed CD8(+) T cells. Previously, two distinct populations of antigen-experienced CD8(+) T cells, termed effector (T(EFF)) and central memory (T(CM)) cells, have been described. After adoptive transfer into CD8-deficient mice, T(EFF), but not T(CM), cells restored AHR, eosinophilic inflammation and IL-13 levels. T(EFF), but not T(CM), cells accumulated in the lungs, and intracellular cytokine staining showed that the transferred T(EFF) cells were a source of IL-13. These data suggest an important role for effector CD8(+) T cells in the development of AHR and airway inflammation, which may be associated with their Tc2-type cytokine production and their capacity to migrate into the lung.

Topics: Adoptive Transfer; Allergens; Alum Compounds; Analysis of Variance; Animals; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; CD8 Antigens; CD8-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Interleukin-13; Lung; Methacholine Chloride; Mice; Mice, Mutant Strains; Ovalbumin; T-Lymphocyte Subsets

Oxidative stress plays an important role in the pathogenesis of bronchial asthma. An excess production of reactive oxygen species (ROS) and defective endogenous antioxidant defense mechanisms may be present in asthma. Reduced glutathione (GSH) is one of the most important reducing agents against oxidant free radicals. A reducing agent, L-2-oxothiazolidine-4-carboxylic acid (OTC), a prodrug of cysteine, increases intracellular GSH. We have used a mouse model for asthma to determine effects of OTC on allergen-induced bronchial inflammation and airway hyper-responsiveness. The administration of OTC reduced bronchial inflammation and airway hyper-responsiveness. ROS generation in bronchoalveolar lavage fluids was increased by ovalbumin (OVA) inhalation, but this increase was diminished by administration of OTC. The increased IL-4, IL-5, IL-13, and eosinophil cationic protein levels in lungs after OVA inhalation were significantly reduced by the administration of OTC. In addition, the increased expression of ICAM-1, VCAM-1, RANTES, and eotaxin in lungs after OVA inhalation was significantly reduced by the administration of OTC. We also showed that the increased NF-kappaB levels in nuclear protein extracts of lung tissues at 72 h after OVA inhalation were decreased by the administration of OTC. These findings suggest that OTC may reduce airway inflammation and hyper-responsiveness through regulation of NF-kappaB activity.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Cell Adhesion Molecules; Chemokines; Disease Models, Animal; Eosinophil Cationic Protein; Interleukins; Lung; Mice; NF-kappa B; Ovalbumin; Prodrugs; Pyrrolidonecarboxylic Acid; Reactive Oxygen Species; Thiazoles; Thiazolidines

To study the possible role of bone marrow-derived hematopoietic cells expressing CD(34)(+) and IL-5 receptor messenger RNA (IL-5R mRNA(+)) in asthmatic airway inflammation.. Balb/c mice were sensitized and challenged by ovalbumin (OVA) to establish the asthmatic model. The control mice were sensitized and exposed to sterile saline. The mice were killed at different time points after challenged by OVA and sterile saline, and bronchoalveolar lavage (BALF), peripheral blood (PB) and bone marrow (BM) were prepared. Eosinophils (EOS) in PB and BALF, and nuclear cells in PB and BM were counted. The percentage of CD(34)(+) cells to nuclear cells (CD(34)(+)%) in PB and BM, and the relative number of CD(34)(+) cells (CD(34)(+)) in PB and BM were calculated by flow cytometry. Immunocytochemistry and in situ hybridization were used to observe the hematopoietic cells with co-localized expression of CD34 and IL-5R mRNA (CD(34)(+)/IL-5R mRNA(+)) in BM. The percentage of BM CD(34)(+)/IL-5R mRNA(+) to BM CD(34)(+) was calculated.. (1) At 6 h after OVA challenge, the number of BALF EOS [(2.67 +/- 1.00) x 105/L] was significantly increased as compared to the number in controls [(0.46 +/- 0.06) x 105/L] (P < 0.01). At 12 h after OVA-challenge, the numbers of BALF EOS [(7.74 +/- 1.98) x 105/L] and PB EOS [(2.91 +/- 0.64) x 108/L] were significantly higher than those in the controls (P < 0.01). At 24 h after OVA-challenge, the numbers of BALF EOS[(19.43 +/- 3.69) x 105/L], PB EOS[(3.93 +/- 0.51) x 108/L] and BM CD(34)(+)/IL-5R mRNA(+) [(300.50 +/- 90.02) per thousand] were increased to the highest levels. The differences were significant as compared to the corresponding parameters in the controls (P < 0.01). At 48 h after OVA-challenge, the numbers of BALF EOS [(12.05 +/- 5.31) x 105/L] and BM CD(34)(+)/IL-5R mRNA(+) [(220.80 +/- 53.41) per thousand] were decreased, but were still significantly different compared to the numbers in the controls (P < 0.01), while other markers returned to the normal levels. (2) The number of BM CD(34)(+)/IL-5R mRNA(+) in the 60 mice was closely correlated with BALF EOS, PB EOS, BM CD(34)(+) and BM CD(34)(+) (P < 0.05).. CD(34)(+) cells expressing IL-5R mRNA, which may favor eosinophilopoiesis and eosinophilic airway inflammation, were increased in the BM of this mouse asthmatic model. A feedback mechanism between the lungs and the bone marrow likely exists, which may be involved in the development and persistence of asthmatic airway inflammation.

Topics: Animals; Antigens, CD34; Asthma; Bone Marrow Cells; Bronchitis; Bronchoalveolar Lavage Fluid; Eosinophils; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Interleukin; Receptors, Interleukin-5; RNA, Messenger

Exposure to endotoxins, allergens, or both in early life might regulate the development of tolerance to allergens later in life.. We investigated whether continuous exposure of infant mice to aerosolized endotoxin, allergen, or both inhibits subsequent allergen-induced immune and inflammatory responses.. Infant BALB/c mice were pre-exposed to aerosolized endotoxin, ovalbumin (OVA), or both (3 times a week for the first 4 weeks of life) before systemic sensitization (days 1-14) and repeated airway challenge (days 28-30) with OVA.. Compared with that seen in negative control animals, systemic sensitization and airway allergen challenges induced high serum levels of allergen-specific IgE (0.7 +/- 0.09 vs 0.02 +/- 0.01 OD units), predominant T(H)2-type cytokine production (IL-5 by splenic mononuclear cells in vitro, 1.2 +/- 0.2 vs 0.04 +/- 0.06 ng/mL), airway inflammation (bronchoalveolar lavage fluid leukocytes, 125 +/- 15 vs 64 +/- 7/microL; eosinophils, 28 +/- 5 vs 1 +/- 0/microL) and development of in vivo airway hyperreactivity (maximal enhanced pause, 11 +/- 1.9 vs 4 +/- 0.2). Pre-exposure with LPS before sensitization increased production of specific IgG2a (67 +/- 10 vs 32 +/- 5 U/mL) but failed to prevent T(H)2-mediated immune responses. Pre-exposure with OVA or with OVA plus LPS completely suppressed allergen sensitization, airway inflammation, and development of in vivo airway hyperreactivity; values were similar to those of negative control animals. Inhibition was due to allergen-specific T-cell anergy indicated by omitted allergen-specific T(H)2 and T(H)1 immune responses. In addition, combined exposure to endotoxin and allergen induced a general shift toward an unspecific T(H)1 immune response.. Exposure with endotoxins before allergen sensitization is not able to induce unresponsiveness but might decrease the susceptibility for sensitization to a variety of common allergens.

Topics: Aerosols; Allergens; Animals; Animals, Newborn; Antibody Formation; Bronchitis; Drug Administration Schedule; Epitopes; Immunization; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Ovalbumin

Interleukin 4 (IL-4) is essential for the switching of B cells to IgE antibody production and for the maturation of T helper (Th) cells toward the Th2 phenotype. These mechanisms are thought to play a crucial role in the pathogenesis of the allergic airway inflammation observed in asthma. In the present study, we examined the anti-inflammatory effects of DNA administration of murine IL-4 mutant Q116D/Y119D (IL-4 double mutant, IL-4DM), which binds to the IL-4 receptor alpha and is an antagonist for IL-4. Immunization of BALB/c mice with alum-adsorbed ovalbumin (OVA) followed by aspiration with aerosolized OVA resulted in the development of allergic airway inflammation. A single administration of IL-4DM DNA before the aerosolized OVA challenge protected the mice from the subsequent induction of allergic airway inflammation. Serum IgE level and extent of eosinophil infiltration in bronchoalveolar lavage (BAL) from IL-4DM DNA-administered mice were significantly lower than those in BAL from control plasmid-immunized mice. In our study, IL-4 or IL-4 mutants were not detected in sera from mice that had received a single administration of IL-4DM DNA. The results of this study provide evidence for the potential utility of IL-4 mutant antagonist DNA inoculation as an approach to gene therapy for asthma.

Topics: Animals; Asthma; Bronchial Provocation Tests; Bronchitis; Bronchoalveolar Lavage Fluid; Cytokines; DNA; Eosinophils; Genetic Therapy; Immunoglobulin E; Interleukin-4; Mice; Mice, Inbred BALB C; Mutation; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Th2 Cells; Vaccines, DNA

Asthma is characterized by chronic airways inflammation, airway wall remodeling, and airway hyperresponsiveness (AHR). An increase in airway smooth muscle has been proposed to explain a major part of AHR in asthma. We have used unbiased stereological methods to determine whether airway smooth muscle hyperplasia and AHR occurred in sensitized, antigen-challenged Brown Norway (BN) rats. Ovalbumin (OA)-sensitized BN rats chronically exposed to OA aerosol displayed airway inflammation and a modest level of AHR to intravenously administered ACh 24 h after the last antigen challenge. However, these animals did not show an increase in smooth muscle cell (SMC) number in the left main bronchus, suggesting that short-lived inflammatory mechanisms caused the acute AHR. In contrast, 7 days after the last aerosol challenge, there was a modest increase in SMC number, but no AHR to ACh. Addition of FCS to the chronic OA challenge protocol had no effect on the degree of inflammation but resulted in a marked increase in both SMC number and a persistent (7-day) AHR. These results raise the possibility that increases in airway SMC number rather than, or in addition to, chronic inflammation contribute to the persistent AHR detected in this model.

Topics: Acetylcholine; Airway Resistance; Animals; Bronchi; Bronchial Hyperreactivity; Bronchitis; Cell Count; Hyperplasia; Hypersensitivity, Immediate; Male; Muscle, Smooth; Ovalbumin; Rats; Rats, Inbred BN

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

Although ebselen, a seleno-organic compound, inhibits inflammation in various animal models, its efficacy as an anti-asthma drug remains to be clarified. In this study, we investigated the inhibitory effect of ebselen on a guinea pig asthma model. Ebselen was orally administered at dosages of 1-20 mg/kg 2 h before an ovalbumin (OA) challenge, and then airway responses, airway inflammation, the generation of superoxide, H(2)O(2), and nitrotyrosine, and the induction of inducible nitric oxide synthase (iNOS) were evaluated. Sensitized animals challenged with OA aerosol showed dual airflow limitations, i.e., immediate and late airway responses (IAR and LAR). Ebselen significantly inhibited LAR at dosages greater than 10 mg/kg, but did not inhibit IAR at any dosage. Bronchoalveolar lavage (BAL) examination showed that airway inflammation was significantly suppressed by ebselen at 10 mg/kg. The generation of superoxide and H(2)O(2) occurred on endothelial cells of LAR bronchi, and was inhibited by 10 mg/kg of ebselen. Superoxide generation was inhibited by diphenyleneiodonium chloride (DPI), a NAD(P)H oxidase inhibitor, but not by allopurinol, a xanthine oxidase inhibitor. Immunoreactivities for iNOS and nitrotyrosine were also observed on endothelial cells of LAR bronchi and were abolished in ebselen-treated animals. The present findings suggest that ebselen can be applied as a new therapeutic agent for asthma. The possible mechanisms by which ebselen inhibits LAR likely involve suppression of oxidant formation and iNOS induction in endothelial cells.

Topics: Airway Resistance; Animals; Antineoplastic Agents, Alkylating; Antioxidants; Area Under Curve; Asthma; Azoles; Bronchitis; Bronchoalveolar Lavage Fluid; Cyclophosphamide; Disease Models, Animal; Endothelium, Vascular; Enzyme Activation; Female; Guinea Pigs; Hydrogen Peroxide; Immunoenzyme Techniques; Isoindoles; Lung; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Organoselenium Compounds; Ovalbumin; Peptide Fragments; Rabbits; Reactive Nitrogen Species; Reactive Oxygen Species; Respiratory Function Tests; Respiratory Hypersensitivity; Superoxides; Tyrosine

CD8+T cells can suppress allergen-induced late airway responses (LARs) and airway inflammation.. To test the hypothesis that the suppression of LARs and airway eosinophilia by CD8+T cells is IFN-gamma mediated, we tested the effects of adoptively transferred CD8+T cells, in which IFN-gamma synthesis was inhibited by an antisense (AS) oligodeoxynucleotide (ODN), on the airway responses of a rat model of allergic asthma.. CD8+T cells were harvested from the cervical lymph nodes of ovalbumin (OVA)-sensitized Brown Norway rats for administration to other actively sensitized syngeneic rats. CD8+T cells (2 x 10(6)) were incubated for 6 hours with 2 micromol/L AS ODN or sense ODN and were injected intraperitoneally into recipients; inhibition of IFN-gamma expression in vitro by AS ODN was shown by means of flow cytometry. Two days later, rats were challenged with aerosolized OVA.. OVA-induced LAR and bronchoalveolar lavage (BAL) fluid eosinophilia were suppressed by sense ODN-treated CD8+T cells. IFN-gamma expression in BAL cells was elevated in these animals. IFN-gamma expression in BAL cells was at control levels in recipients of AS ODN-treated CD8+ cells, confirming the success of the AS treatment in vivo. BAL eosinophilia was also largely restored in the AS ODN treatment group. In contrast, the CD8+T cell-induced suppression of the LAR was not significantly affected by AS ODN pretreatment.. These results indicate that CD8+T cells inhibit airway eosinophilia through secretion of IFN-gamma but may suppress the LAR by means of other mechanisms.

Topics: Adoptive Transfer; Allergens; Animals; Bronchi; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; CD8-Positive T-Lymphocytes; Eosinophilia; Interferon-gamma; Male; Oligonucleotides, Antisense; Ovalbumin; Rats; Rats, Inbred BN; Time Factors

1. Recently, much attention has been paid to the relationship between the nervous and immune systems. The present study was conducted to clarify the role of neurotrophin low affinity receptor (p75N) in allergic airway inflammation and hyper-responsiveness (AHR) in mice by employing p75N gene deficient mice. 2. Mice were immunized twice by intraperitoneal injections of ovalbumin (OA) at intervals of 12 days. OA was inhaled 10 days after the secondary immunization and repeated three times at 4 days interval. Twenty-four hours after the last inhalation, airway responsiveness to acetylcholine was measured and bronchoalveolar lavage fluid (BALF) was obtained for examining the number of inflammatory cells and the level of cytokines. Serum immunoglobulin was measured as a marker of systemic immune response before the final inhalation. 3. In wild-type mice, repeated antigen provocation resulted in airway eosinophilia, AHR and elevations in serum IgE and interleukin (IL)-4 and -5 in BALF. In p75N gene deficient mice, none of the above parameters was observed after antigen provocation. The antigen-induced production of interferon (IFN)-gamma and nerve growth factor (NGF) were not altered by depletion of p75N gene. 4. The present findings suggest that p75 gene deficiency disrupt an allergic airway inflammation and AHR in mice by interfering type 2 helper T (Th2) cell responses.

Topics: Acetylcholine; Animals; Antigens; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cytokines; Dose-Response Relationship, Drug; Genotype; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Nerve Growth Factor; Ovalbumin; Receptor, Nerve Growth Factor; Receptors, Nerve Growth Factor; Vasodilator Agents

Brown-Norway (BN) rats develop airway hyper-responsiveness and lung eosinophilia 18-24 h after ovalbumin (OA) challenge. We hypothesized therefore that allergen-induced airway inflammation would further enhance airway responses to a subsequent antigen challenge. Animals were sensitized to both OA and bovine serum albumin (BSA) and, 14 days later, challenged by aerosols with both antigens 24 h apart. Measurements of pulmonary resistance (RL) were made for 8 h after the second antigen challenge and bronchoalveolar lavage (BAL) was performed. Animals were divided into three groups and received two challenges as follows: saline-BSA (n=9), OA-saline (n=8) and OA-BSA (n=10). Sensitization was confirmed by measurements of specific OA-IgE and BSA-IgE. Early responses [determined as the highest value of RL within the first 30 min after the challenge] were absent in all study groups. The late responses [determined from the area under the RL versus time curve from 120 to 480 min after the challenge] were significantly greater in animals challenged with BSA (15.16+/-3.86) compared to saline (3.76+/-4.09; P<0.05). However previous exposure to OA did not further increase the late response in animals subsequently challenged with BSA (20.11+/-3.67) despite enhanced airway responsiveness to LTD4 at this time point. BAL eosinophils and lymphocytes were significantly increased following BSA challenge in previously OA-challenged animals, compared to numbers retrieved from animals previously exposed to saline (P<0.05). These data indicate that previous exposure to OA did not further increase the LR to a second antigen challenge despite substantial increases in airway inflammatory cells and airway hyper-responsiveness to LTD4.

Topics: Animals; Antigens; Bronchi; Bronchial Provocation Tests; Bronchitis; Bronchoalveolar Lavage Fluid; Immunoglobulin E; Leukotriene D4; Male; Ovalbumin; Rats; Serum Albumin

To observe the time-course profile of airway inflammation in murine asthma model, and to analyze the dynamic profile of pro-inflammatory factor IL-5, and inflammtion-suppressing factor IL-10 in the airway.. An animal model of asthma was established by OVA sensitizing-challenging BALB/C mice. At seven points (0 h, 8 h, 24 h, 48 h, 96 h, days 7 and days 14) in the time course after challenge, bronchoalveolar lavage (BAL) was performed. Total cell counts, cell differential, eosinophil (EOS) counts and percentages were examined and levels of two Th2 cytokines, IL-5 and IL-10, were detected by ELISA.. After OVA challenge, total cell counts, EOS counts and EOS percentages in BALF were increased significantly. EOS count increased at 8 h, formed a plateau from 24 h to 48 h, declined at 96 h, reduced approximately to control from days 7 to days 14. IL-5 in BALF elevated at 8 h, maitained at high level from 28 h to 96 h, decreased at days 7. The dynamic changes of IL-5 was in synchronism with that of EOS counts, a positive correlation being identified. No significant difference was found between IL-10 in BALF before and after challenge. A close correlation between EOS counts and IL-5/IL-10 ratio was demonstrated (r = 0.9, P < 0.001).. IL-5 and IL-10 up- and down-regulate asthmatic airway inflammation respectively, and their interaction may play an important role in inflammatory progress and resolution.

Topics: Animals; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Interleukin-10; Interleukin-5; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Time Factors

Sensitized Brown Norway rats are known to develop eosinophilic bronchial inflammation and airway hyperresponsiveness after Ag exposure. However, we have previously observed that sensitized aged rats of the same strain failed to develop such allergic inflammation. In the present study, we investigated age-associated changes of cytokine mRNA expression in bronchoalveolar lavage (BAL) cells. Both young (8- to 10-wk-old) and aged (100- to 120-wk-old) Brown Norway rats were sensitized with OVA, and BAL was performed 24 h after OVA inhalation challenge. Semiquantitative RT-PCR analysis of BAL cells showed that the cells from aged rats preferentially expressed Th1 type cytokine (IFN-gamma) mRNA, whereas cells from young animals expressed more Th2 type cytokine mRNAs including those for IL-4 and IL-5. Decreased expression of Th2 type cytokine transcripts in aged animals was further confirmed by quantitative analysis, competitive RT-PCR of BAL cells, and in situ hybridization. The age-associated changes of cytokine profile were not restricted to BAL cells but were a general feature of lymphocytes, as shown by examination of popliteal lymph nodes draining the site of sensitization. These findings suggest that decreased allergic inflammation in aged animals is attributable to age-dependent impairment of Th2 generation in response to Ag.

Topics: Age Factors; Aging; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Cell Differentiation; Cell-Free System; Cells, Cultured; Cytokines; Enzyme-Linked Immunosorbent Assay; Injections, Subcutaneous; Interferon-gamma; Interleukin-4; Interleukin-5; Lymph Nodes; Male; Ovalbumin; Rats; Rats, Inbred BN; Respiratory Hypersensitivity; RNA, Messenger; Th2 Cells; Time Factors

Using a guinea pig model of allergic asthma, we investigated the effects of the inhaled, highly selective nonpeptide tachykinin NK1 and NK2 receptor antagonists SR 140333 and SR 48968, respectively, on allergen-induced early (EAR) and late (LAR) asthmatic reactions, airway hyperreactivity (AHR) after these reactions, and infiltration of inflammatory cells in the airways. Both SR 140333 (100 nM, 3 min) and SR 48968 (100 nM, 3 min) had no effect on the severity of the EAR, while the NK2 receptor antagonist SR 48968, but not the NK1 receptor antagonist SR 140333, caused significant inhibition of the LAR. SR 140333 significantly reduced the allergen-induced AHR to histamine, both after the EAR and the LAR. By contrast, SR 48968 did not affect the AHR after the EAR, but significantly attenuated the AHR after the LAR. Bronchoalveolar lavage studies performed after the LAR indicated that SR 140333 caused significant inhibition of allergen-induced infiltration of eosinophils, neutrophils and lymphocytes, while SR 48968 attenuated the infiltration of neutrophils and lymphocytes, but not of eosinophils. Both NK receptor antagonists tended to reduce the accumulation of ciliated epithelial cells in the airways. These results indicate that NK1 and NK2 receptors are importantly, but differentially, involved in the development of allergen-induced airways obstruction, AHR and infiltration of inflammatory cells in the airways. Therefore, both NK1 and NK2 receptor antagonists, or dual NK1 and NK2 antagonists, could be useful in the treatment of allergic asthma.

Topics: Allergens; Animals; Asthma; Benzamides; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchitis; Chemotaxis, Leukocyte; Eosinophils; Guinea Pigs; Lymphocytes; Neurokinin-1 Receptor Antagonists; Neutrophils; Ovalbumin; Piperidines; Quinuclidines; Receptors, Neurokinin-1; Receptors, Neurokinin-2

Asthma and its exacerbation by air pollution are major public health problems. This investigation sought to more precisely model this disorder, which primarily affects children, by using very young mice. The study first attempted to create allergic airway hypersensitivity in neonatal mice and to determine if physiologic testing of airway function was possible in these small animals. Neonatal mice were sensitized by i.p. injection of ovalbumin (OVA, 5 microg) and alum (1 mg) at 3 and 7 d of age. One week later, mice were challenged by allergen nebulization (3% OVA in PBS, 10 min/d, d 14-16). OVA-exposed mice showed: (1) increased airway hyperresponsiveness (AHR) to methacholine by whole-body plethysmography; (2) eosinophilia in bronchoalveolar lavage (BAL) fluid; (3) airway inflammation using histopathology techniques; and (4) elevated serum anti-OVA immunoglobulin E. Hence, these neonatal mice were successfully sensitized and manifested "asthmatic" responses after allergen challenge. Experiments were conducted to investigate the effect of one surrogate for ambient air particles, residual oil fly ash (ROFA), on this juvenile asthma model. Aerosolized ROFA leachate (supernatant of 50 mg/ml, 30 min, on d 15) had no marked effect alone, but caused a significant increase in AHR and airway inflammation in OVA-sensitized and challenged mice. This synergistic effect was abrogated by the antioxidant dimethylthiourea (DMTU, 3 mg/kg mouse, i.p.). This model may be useful to study air pollution-mediated exacerbation of asthma in children.

Topics: Aerosols; Air Pollutants; Allergens; Animals; Animals, Newborn; Asthma; Bronchial Hyperreactivity; Bronchitis; Carbon; Coal Ash; Enzyme-Linked Immunosorbent Assay; Free Radical Scavengers; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Phenotype; Plethysmography; Serine Proteinase Inhibitors; Thiourea

Using a guinea pig model of acute allergic asthma, we recently established that a deficiency of nitric oxide (NO) contributes to airway hyperreactivity (AHR) after the early asthmatic reaction (EAR) and that restoration of NO activity may contribute to the (partial) reversal of AHR after the late asthmatic reaction (LAR). In the present study, we investigated the role of iNOS-derived NO in the regulation of AHR to histamine after the LAR. Inhalation of a selective dose of the specific iNOS inhibitor aminoguanidine (0.1 mM, 3 min) had no effect on basal airway reactivity to histamine in unchallenged, ovalbumin-sensitized animals and did not affect the allergen-induced AHR after the EAR. By contrast, this dose of aminoguanidine significantly potentiated the partially reduced AHR after the LAR to the level of AHR observed after the EAR, indicating that induction of iNOS during the LAR contributes to the reversal of AHR. Inhalation of a higher aminoguanidine concentration (2.5 mM) shortly before the onset of the LAR diminished the AHR after the LAR and reduced the number of neutrophils, lymphocytes, and ciliated epithelial cells in the bronchoalveolar lavage at this time point. The results indicate that iNOS-derived NO may have both beneficial and detrimental effects on allergen-induced AHR to histamine after the LAR by functional antagonism of histamine-induced bronchoconstriction, and by promoting airway inflammation and epithelial damage on the other hand, respectively.

Topics: Acute Disease; Adjuvants, Immunologic; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cell Count; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme Inhibitors; Epithelial Cells; Female; Guanidines; Guinea Pigs; Histamine; Leukocyte Count; Lymphocyte Count; Male; Neutrophils; Nitric Oxide; Nitric Oxide Synthase; Ovalbumin

To investigate the relations between intrinsic nitric oxide (NO) and airway inflammation and lymphocyte proliferation in bronchial asthma.. The rat model of asthmatic airway inflammation was established by first sensitizing and then challenging the animals with ovalbumin. NO synthase (NOS) inhibitor and NO precursor were then applied and their influences on the airway inflammatory cell numbers and on the numbers of mIL-2R positive lymphocytes were observed.. In vivo experiments showed that normal control rats (n = 6) did not have eosinophils in their submucosal of airways while in the sensitized animals (n = 6) eosinophils [23 +/- 5 (average cell counts per microscopic visual field, the same below) as well as lymphocytes (34 +/- 5) and mIL-2R positive cells (12 +/- 3) were found in significantly increased numbers in the airways. The application of NOS inhibitor significantly reduced eosinophils (13 + 3, P < 0.05) and mIL-2R positive cells (4.3 +/- 1.6, P < 0.05) in the sensitized animals and the number of lymphocytes was also decreased (28 +/- 4) although it did not reach the significant level. At the same time the spleen cell proliferation was inhibited (P < 0.05) and the mIL-2R positive spleen cell numbers were reduced (P < 0.05). On the contrary, the application of NO precursor resulted in the further increase of the numbers of eosinophils, lymphocytes and mIL-2R positive cells although the differences were not statistically significant. In vitro experiments showed that NOS inhibitor inhibited the proliferation of cultured spleen lymphocytes as well as their mIL-2R expression (P < 0.05). NO precursor, when used in low dose, could promote the proliferation and mIL-2R expression of spleen cells (P < 0.05); but high dose of it showed inhibiting effect on the spleen cells (P < 0.05).. Intrinsic NO at a suitable level is important in the regulation of the proliferation and activation of lymphocytes in the airways with allergic inflammation in the sensitized rats. This research is conducive to the understanding of the pathogenesis of the asthmatic airway inflammation and to investigating of new therapeutic approaches.

Topics: Animals; Arginine; Asthma; Bronchitis; Cells, Cultured; Eosinophils; Lymphocytes; Male; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Ovalbumin; Random Allocation; Rats; Rats, Sprague-Dawley; Receptors, Interleukin-2

The effects of IFN-beta and prednisolone (Pred) on antigen-induced IgE antibody production, airway eosinophilia and airway hyperreactivity (AHR) were studied in ovalbumin-sensitized Balb/c mice. Three inhalations of antigen (ovalbumin) caused an increase in the number of eosinophils and lymphocytes in bronchoalveolar lavage fluid and AHR to acetylcholine with a significant elevation in the serum IgE level. IFN-beta clearly inhibited the antigen-induced airway inflammation and AHR, but did not affect IgE antibody production. Pred inhibited antigen-induced IgE antibody production, airway inflammation and AHR to acetylcholine. In addition, IFN-beta inhibited T-helper type 2 (Th2) cell clone (D10G4.1)-induced peritoneal eosinophilia in mice, but did not affect neutrophilia, whereas Pred inhibited D10G4.1-induced peritoneal eosinophilia and neutrophilia. These results suggest that IFN-beta inhibits antigen-induced bronchial inflammation and AHR probably due to the inhibition of Th2-induced airway eosinophilia.

Topics: Acetylcholine; Adjuvants, Immunologic; Administration, Inhalation; Animals; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage Fluid; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Immunoglobulin E; Immunoglobulin G; Interferon-beta; Leukocyte Count; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Peritoneal Cavity; T-Lymphocytes

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

Guinea-pigs sensitized by a subcutaneous injection of ovalbumin in Al(OH)3 and boosted 2 weeks later exhibit marked bronchial hyperresponsiveness to various agonists and intense bronchial wall infiltration by CD4+ T-lymphocytes and eosinophils. We have compared the effect of FK506, a novel immunosuppressive agent, on the mucosal infiltration by T-cells and eosinophils with the well established drugs, nedocromil sodium and dexamethasone. Sensitized Hartley guinea-pigs were treated subcutaneously for 5 days with FK506 (100 micrograms.kg-1 daily), nedocromil sodium (30 micrograms.kg-1 daily), or dexamethasone (200 micrograms.kg-1 daily). On the day of the experiment, i.e. one week after the booster injection of antigen, the animals were killed, the lungs dissected, frozen and cryostat sections stained by immunohistochemical methods using monoclonal antibodies specific for total T-lymphocytes, CD4+ and CD8+ T-cells. Cyanide-resistant eosinophil peroxidase activity was used to stain the eosinophils. Sections were coded and positive cells enumerated in the lamina propria and adventitia of the bronchi. Sensitized and antigen-stimulated vehicle-treated guinea-pigs showed marked infiltration of the bronchial wall by CD4+ T-lymphocytes and eosinophils compared with sensitized, non-antigen stimulated animals. As compared to vehicle, FK506 or dexamethasone abolished the T-cell/eosinophil invasion in the bronchial wall, whereas nedocromil sodium was ineffective in protecting the lungs from T-lymphocyte or eosinophil infiltration. We conclude that both FK506 and dexamethasone are effective in curtailing bronchial inflammation in allergic guinea-pigs, whereas nedocromil sodium did not resolve the inflammation associated with T-lymphocytes or eosinophils.

Topics: Animals; Anti-Inflammatory Agents; Antigens; Bronchi; Bronchitis; Dexamethasone; Eosinophil Peroxidase; Eosinophils; Guinea Pigs; Immunization; Inflammation; Lung; Male; Mucous Membrane; Nedocromil; Ovalbumin; Peroxidases; Respiratory Hypersensitivity; T-Lymphocytes; Tacrolimus

The causative relationship between airway inflammation and hyperreactivity is unclear, since inflammatory changes have been examined at one or, at most, a few time-points after antigen challenge in both human asthma and animal models. We have made a detailed investigation of inflammatory and functional changes in the airways up to 8 days after antigen challenge in guinea-pigs. In particular, we examined the hypothesis that eosinophil-derived mediators contribute to tissue damage and the development of airway hyperresponsiveness. Following antigen challenge, the influx of inflammatory cells and mediator release in airway tissue and bronchoalveolar lavage fluid were correlated temporally with histopathological changes in airway tissue and airway responsiveness. Eosinophil influx was demonstrable at 4 h. Eosinophilia peaked after 24 h and persisted for at least 8 days. Parallel increases in the concentrations of major basic protein and eosinophil cationic protein in bronchoalveolar lavage fluid indicated that the eosinophils were activated. Eosinophilia was accompanied by subepithelial oedema and epithelial damage co-localized with major basic protein immunoreactivity. A transient neutrophilia (< 48 h duration) and an increase in neutrophil elastase in bronchoalveolar lavage fluid peaked at 14 h. The proportion of airway macrophages with an activated morphology increased at 8 h and remained markedly elevated until 72 h. Airways were hyperresponsive to histamine at 4 h and for at least 8 days. The antigen-induced airway inflammation resemble in time-course and histopathology that seen in antigen-challenged asthmatics, and indicate that the eosinophil and its cytotoxic proteins may be major mediators of airway mucosal damage and airway hyperresponsiveness.

Topics: Aluminum Hydroxide; Animals; Blood Proteins; Bronchial Hyperreactivity; Bronchitis; Bronchoalveolar Lavage; Eosinophil Granule Proteins; Eosinophils; Guinea Pigs; Immunization; Leukocyte Elastase; Lung; Lymphocytes; Macrophages; Male; Neutrophils; Ovalbumin; Pancreatic Elastase; Ribonucleases; Time Factors

In a new model using conscious, unrestrained and ovalbumin-sensitized guinea pigs, we investigated the effects of the selective histamine H1 receptor antagonist, mepyramine, on the development of allergen-induced early and late asthmatic reactions, bronchial hyperreactivity and airway inflammation, having each animal as its own control. In guinea pigs responding to a first allergen exposure with an early as well as a late asthmatic reaction (82% of the animals) a second, identical, allergen provocation was performed, in the absence (control) or presence of 1 mg/ml mepyramine aerosol, inhaled for 10 min, 1 h before provocation. The mepyramine treatment significantly reduced both early and late asthmatic reactions and prevented the development of bronchial hyperreactivity to histamine and methacholine after both reactions. Examination of the bronchoalveolar lavage fluid 24 h after the second allergen provocation revealed a general reduction of inflammatory cells after mepyramine treatment. The results indicate that histamine, released during the early asthmatic reaction, contributes to the development of the late asthmatic reaction as well as of early and late bronchial hyperreactivity, possibly via an effect on airway inflammation.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchitis; Bronchoalveolar Lavage Fluid; Guinea Pigs; Histamine; Male; Methacholine Compounds; Ovalbumin; Pyrilamine; Respiratory Function Tests

Exposure of non-sensitized Brown Norway (BN) rats to a 10%-ovalbumin aerosol induced an increase in the number of neutrophils in the broncho-alveolar lavage (BAL) fluid 3 and 6 h later but with no change in number of cells at 24 h. When the BN rats were actively sensitized (i.m. injection of 10 mg/kg ovalbumin and i.p. injection of killed Bordetella pertussis) and exposed 12-14 days later to a 10%-ovalbumin aerosol there was an increase in the number of eosinophils in the BAL fluid, maximal 24-48 h after the anaphylactic reaction. The increase in the number of neutrophils in the bronchial lumen 3 and 6 h after the anaphylactic reaction was larger than that obtained in non-specific inflammation and in contrast to this was still present 24-48 h after ovalbumin exposure. In passively sensitized BN rats exposed to ovalbumin aerosol, no inflammation appeared in the BAL fluid 24 h after the anaphylactic reaction. Various drugs, administered twice, 5 min and 5 h after the anaphylactic reaction, have been evaluated for their effects on the 24-h inflammation obtained in actively sensitized rats. Dexamethasone acetate (0.08 mg/kg i.p.) and theophylline (50 mg/kg i.p.) decreased the number of eosinophils and neutrophils. Ketotifen fumarate (12.5 mg/kg), cetirizine dihydrochloride (12.5 mg/kg), salbutamol (2 mg/kg), disodium cromoglycate (50 mg/kg) all given intraperitoneally, reduced the number of eosinophils. Tioxamast decreased the number of eosinophils at 12.5 mg/kg i.p. and by the oral route.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Albuterol; Animals; Bronchitis; Dexamethasone; Disease Models, Animal; Ketotifen; Male; Ovalbumin; Rats; Rats, Inbred BN; Respiratory Hypersensitivity; Theophylline

The action of theophylline was studied on the inflammatory reaction obtained in bronchoalveolar lavage fluid 24 h after an active anaphylactic shock had been induced by ovalbumin inhalation in conscious sensitized guinea-pigs. The compound was administered twice by intraperitoneal administration after the anaphylactic reaction at a dose of 50 mg kg-1. When the guinea-pigs were sensitized by intramuscular injection of 30 mg kg-1 ovalbumin or by ovalbumin aerosol, theophylline reduced the number of eosinophils and mononuclear cells in the fluid. When animals were sensitized by intramuscular injection of 30 mg kg-1 ovalbumin mixed with Freund's complete adjuvant, treatment with the xanthine derivative decreased only the number of eosinophils. In the three models theophylline did not modify significantly the number of neutrophils. Thus theophylline always reduced pulmonary eosinophilia irrespective of the mode of sensitization used to induce anaphylactic shock.

Topics: Aerosols; Anaphylaxis; Animals; Bronchitis; Bronchoalveolar Lavage Fluid; Eosinophils; Guinea Pigs; Injections, Intramuscular; Lung; Male; Neutrophils; Ovalbumin; Theophylline

Twenty-four hours after an active anaphylactic shock induced by inhalation of antigen in conscious guinea pigs sensitized by a large dose of ovalbumin in complete Freund's adjuvant, a noteworthy bronchial inflammation, characterized by increased numbers of neutrophils, mononuclear cells and eosinophils in the bronchoalveolar lavage fluid, was observed. Some drugs administered after the anaphylactic shock were investigated using this model. Disodium cromoglycate primarily reduced the number of mononuclear cells and eosinophils. Dexamethasone and theophylline decreased the number of eosinophils. Salbutamol and mepyramine increased neutrophils. Indomethacin did not give rise to any significant effect. This test appears to be of use for the investigation of anti-inflammatory compounds in the prophylactic treatment of asthma.

Topics: Albuterol; Anaphylaxis; Animals; Asthma; Bronchitis; Bronchoalveolar Lavage Fluid; Cromolyn Sodium; Dexamethasone; Disease Models, Animal; Guinea Pigs; Male; Ovalbumin; Pyrilamine

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

Topics: Animals; Bronchitis; Calcium; Chickens; Egg Yolk; Female; Hydrogen-Ion Concentration; Magnesium; Ovalbumin; Oviducts; Ovulation; Ovum; Potassium; Poultry Diseases; Proteins; Sodium; Virus Diseases