ovalbumin and Bronchial-Diseases

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

Allergen avoidance and anti-inflammatory therapy are standard therapeutic approaches guidelines advocate to control asthma symptoms. Currently, it is not known whether such strategies reduce airway remodeling.. We have therefore used a mouse model of allergen-induced airway remodeling to determine whether allergen avoidance combined with corticosteroid therapy can reverse established airway remodeling.. Mice were sensitized to ovalbumin and then repetitively challenged with intranasal ovalbumin for 3 months to develop structural features of airway remodeling including peribronchial fibrosis and increased thickness of the peribronchial smooth muscle layer. At this time point, mice were treated with allergen avoidance, allergen avoidance and corticosteroids, or corticosteroids for 1 month to determine whether either strategy could reverse established airway remodeling.. Mice repetitively challenged with ovalbumin developed peribronchial fibrosis (increased total lung collagen and increased peribronchial trichrome staining) as well as increased thickness of the peribronchial smooth muscle layer. Allergen avoidance significantly reduced airway inflammation and mucus expression, slightly reduced peribronchial fibrosis, and had no effect on the thickness of the peribronchial smooth muscle layer. Addition of corticosteroids to allergen avoidance significantly reduced levels of peribronchial fibrosis as well as the thickness of the peribronchial smooth muscle layer.. Allergen avoidance reduces airway inflammation and mucus expression but has more limited immediate effects on reducing structural features of established airway remodeling. The combination of allergen avoidance and corticosteroid therapy is effective in reversing established features of airway remodeling including peribronchial fibrosis and the increased thickness of the smooth muscle layer.

Topics: Administration, Intranasal; Adrenal Cortex Hormones; Animals; Bronchi; Bronchial Diseases; Drug Administration Schedule; Environment, Controlled; Female; Fibrosis; Mice; Mice, Inbred BALB C; Mucus; Muscle, Smooth; Ovalbumin; Time Factors; Transforming Growth Factor beta

In response to inflammation or injury, airway epithelial cells express inducible genes that may contribute to allergen-induced airway remodeling. To determine the contribution of epithelial cell NF-kappaB activation to the remodeling response, we generated CC10-Cre(tg)/Ikkbeta(delta/delta) mice in which NF-kappaB signaling through IkappaB kinase beta (IKKbeta) is selectively ablated in the airway epithelium by conditional Cre-recombinase expression from the Clara cell (CC10) promoter. Repetitive ovalbumin challenge of mice deficient in airway epithelial IKKbeta prevented nuclear translocation of the RelA NF-kappaB subunit only in airway epithelial cells, resulting in significantly lower peribronchial fibrosis in CC10-Cre(tg)/Ikkbeta(delta/delta) mice compared with littermate controls as assessed by peribronchial trichrome staining and total lung collagen content. Levels of airway mucus, airway eosinophils, and peribronchial CD4+ cells in ovalbumin-challenged mice were also reduced significantly upon airway epithelial Ikkbeta ablation. The diminished inflammatory response was associated with reduced expression of NF-kappaB-regulated chemokines, including eotaxin-1 and thymus- and activation-regulated chemokine, which attract eosinophils and Th2 cells, respectively, into the airway. The number of peribronchial cells expressing TGF-beta1, as well as TGF-beta1 amounts in bronchoalveolar lavage, were also significantly reduced in mice deficient in airway epithelium IKKbeta. Overall, these studies show an important role for NF-kappaB regulated genes in airway epithelium in allergen-induced airway remodeling, including peribronchial fibrosis and mucus production.

Topics: Active Transport, Cell Nucleus; Allergens; Animals; Bronchial Diseases; CD4-Positive T-Lymphocytes; Cell Nucleus; Cytokines; Eosinophils; Epithelium; Fibrosis; Gene Deletion; Genotype; I-kappa B Kinase; Leukocyte Count; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mucus; Muscle, Smooth; NF-kappa B; Ovalbumin; Promoter Regions, Genetic

Release of human lung mast cell tryptase may be important in the pathophysiology of asthma. We examined the effect of the reversible, nonelectrophilic tryptase inhibitor MOL 6131 on airway inflammation and hyper-reactivity in a murine model of asthma. MOL 6131 is a potent selective nonpeptide inhibitor of human lung mast cell tryptase based upon a beta-strand template (K(i) = 45 nM) that does not inhibit trypsin (K(i) = 1,061 nM), thrombin (K(i) = 23, 640 nM), or other serine proteases. BALB/c mice after i.p. OVA sensitization (day 0) were challenged intratracheally with OVA on days 8, 15, 18, and 21. MOL 6131, administered days 18-21, blocked the airway inflammatory response to OVA assessed 24 h after the last OVA challenge on day 22; intranasal delivery (10 mg/kg) had a greater anti-inflammatory effect than oral delivery (10 or 25 mg/kg) of MOL 6131. MOL 6131 reduced total cells and eosinophils in bronchoalveolar lavage fluid, airway tissue eosinophilia, goblet cell hyperplasia, mucus secretion, and peribronchial edema and also inhibited the release of IL-4 and IL-13 in bronchoalveolar lavage fluid. However, tryptase inhibition did not alter airway hyper-reactivity to methacholine in vivo. These results support tryptase as a therapeutic target in asthma and indicate that selective tryptase inhibitors can reduce allergic airway inflammation.

Topics: Animals; Asthma; Bridged Bicyclo Compounds, Heterocyclic; Bronchial Diseases; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Movement; Cytokines; Eosinophils; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Models, Molecular; Mucus; Ovalbumin; Piperidines; Pulmonary Edema; Pulmonary Eosinophilia; Serine Endopeptidases; Serine Proteinase Inhibitors; Tryptases; Vascular Cell Adhesion Molecule-1

The influence of stress and diazepam treatment on airway inflammation was investigated in ovalbumin (OVA)-sensitized rats. Animals were injected with OVA plus aluminum hydroxide intraperitoneally (day 0) and boosted with OVA subcutaneously (day 7). From the first to 13th day after sensitization, rats were treated with diazepam, and 1 h later they were placed in a shuttle box where they received 50 mild escapable foot shocks/day preceded by a sound signal (S). Response during the warning (S) canceled shock delivery and terminated the S. On day 14, rats were submitted to a single session of 50 inescapable foot shocks preceded by S and then were challenged with OVA. High levels of stress were detected in shocked animals, manifested as ultrasonic vocalizations. Morphometric analysis of stressed animals revealed a significant increase in both edema and lymphomononucleated cells in airways compared with controls. Diazepam treatment reduced edema in stressed and nonstressed rats. No differences were found in polymorphonucleated cell infiltration. Diazepam treatment reduced lymphomononucleated cell infiltration in stressed animals. These data suggest that stress and diazepam treatment play relevant roles in edema and lymphomononucleated airway inflammation in OVA-sensitized rats.

Topics: Administration, Inhalation; Anaphylaxis; Animals; Anti-Anxiety Agents; Bronchial Diseases; Conditioning, Psychological; Diazepam; Edema; Electroshock; Injections, Intraperitoneal; Lung; Male; Ovalbumin; Pneumonia; Rats; Rats, Wistar; Specific Pathogen-Free Organisms; Stress, Physiological

The relationship between CD4(+) T cell-mediated airway eosinophilic inflammation and bronchial hyper-responsiveness (BHR) was investigated. Ovalbumin-reactive T(h)0 clones were adoptively transferred to unprimed BALB/c mice and then the mice were challenged by inhalation of the relevant antigen. Upon antigen provocation, infused T(h) clones infiltrated into the airways, followed by the accumulation and degranulation of eosinophils, goblet cell hyperplasia, edema and increase in bronchial responsiveness to acetylcholine. Transfer of several clones that differed in the levels of IL-5 production revealed that the magnitude of in vivo eosinophilia strongly correlated with the IL-5-producing capacity of the infused T(h) clones. Administration of anti-IL-5 mAb almost completely suppressed antigen-induced eosinophilic inflammation and BHR. Administration of anti-IL-4 mAb or anti-IFN-gamma mAb enhanced the eosinophilia and BHR, whereas anti-IL-2 mAb did not affect them. The number of accumulated eosinophils significantly correlated with the intensity of BHR. Our present results clearly demonstrated that CD4(+) T cells induced BHR as a result of eosinophilic inflammation. IL-5 totally regulated both responses.

Topics: Acetylcholine; Animals; Antibodies, Monoclonal; Bronchial Diseases; Bronchial Hyperreactivity; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Clone Cells; Cytokines; Eosinophil Peroxidase; Eosinophilia; Interferon-gamma; Interleukin-4; Interleukin-5; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Peroxidases; Pulmonary Eosinophilia; Specific Pathogen-Free Organisms

The role of small airways in the immediate allergic response is largely unknown. We therefore used the model of precision-cut lung slices (PCLS) in combination with quantitative videomicroscopy to study the early allergic response to allergen in airways ranging from 50 to 900 microm. After PCLS from untreated Wistar rats had been passively sensitized for 16 h with serum from sensitized Brown Norway rats, exposure to 0.1% ovalbumin resulted in an immediate allergic response. Both extent (r = 0.74, p < 0.0001) and velocity (r = 0.49, p < 0.0001) of the allergen-induced bronchoconstriction increased with decreasing airway size. In addition, we observed that smaller airways not only contracted stronger and quicker, but that they also relaxed faster, suggesting that smaller airways are more reactive in principle. The allergen-induced bronchoconstriction in PCLS was prevented by the serotonin receptor antagonist ketanserin (IC(50) 6 nM), but not by antagonists directed against histamine, acetylcholine, PAF, or endothelin receptors, or by cyclooxygenase or lipoxygenase inhibitors. Like allergen, serotonin provoked responses that were stronger in smaller airways. These findings suggest that the immediate allergic response in rat PCLS depends largely on serotonin and that this response can occur in nearly all airway generations, but is most pronounced in the smallest airways, that is, the terminal bronchioles.

Topics: Allergens; Animals; Asthma; Bronchial Diseases; Constriction, Pathologic; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Hypersensitivity, Immediate; Immunization; Ketanserin; Microscopy, Video; Ovalbumin; Rats; Rats, Wistar; Serotonin; Serotonin Antagonists

Effects of U-67590A, an analogue of methylprednisolone, on antigen-induced bronchoconstriction expressed as ventilation overflow were examined in ovalbumin-sensitized guinea pigs. When administered intravenously 17-18 hr before the challenge with antigen, U-67590A at doses of 10 and 30 mg/kg caused dose-dependent inhibition of increased ventilation overflow immediately after challenge. Death due to anaphylactic shock was markedly reduced by U-67590A. At 10 mg/kg (i.v.), U-67590A given 10 min before the challenge significantly inhibited the antigen-induced increase in ventilation overflow; the greatest inhibition seen 5-6 hr prior to the challenge. Pretreatment with 10 mg/kg FPL-55712 attenuated the increase in ventilation overflow during anaphylaxic shock. It is conceivable that inhibition of the leukotriene-mediated response is involved in the glucocorticoid-induced suppression of airway obstruction in anaphylaxis, and that inhibitory action of the glucocorticoid directly acting on the airway may account for the very fast onset of action.

Topics: Anaphylaxis; Animals; Antigens; Asthma; Bronchial Diseases; Chromones; Constriction, Pathologic; Dose-Response Relationship, Drug; Guinea Pigs; Male; Methylprednisolone; Ovalbumin; SRS-A; Time Factors

Intravenous application of substance P (SP) (10 micrograms/kg) caused bronchoconstriction returning to baseline within 10 min. Pretreatment with SP did not influence the bronchoconstrictor response induced by allergen or carbachol.

Topics: Allergens; Animals; Bronchi; Bronchial Diseases; Carbachol; Constriction, Pathologic; Female; Guinea Pigs; Male; Ovalbumin; Substance P

Respiratory histaminergic and cholinergic receptor function was investigated in isolated tracheal spirals of guinea pigs receiving different diets. Comparison was made between saline treated (controls) and Haemophilus influenzae treated animals in non sensitized conditions, the latter being a model for bronchial hyperreactivity, and in sensitized conditions, being a model for allergen induced bronchial hypersensitivity. The different semi-synthetic diets (35 energy% fat), varying in linoleic acid content (5.85, 11.25 and 22.05 en% fat) and one diet with low linoleic acid (3.55 en%) in which linolenic acid was added additionally (5.30 en%), exerted profound effects on tracheal reactivity to histamine. In sensitized animals the maximal induced histamine contraction was significantly diminished in the dietary group receiving 5.85 en% linoleic acid as compared with the other dietary groups (35% decrease in the H. influenzae-treated, 20-30% decrease in saline treated animals). Results in non-sensitized animals were similar, though less pronounced. No effect on food intake or growth of the animals could be demonstrated during the six week experimental periods. The carbachol induced contraction of the tracheal spirals of sensitized animals receiving low linoleic acid was also significantly decreased as compared to the other dietary groups (30% for saline treated and 20-30% for H. influenzae-treated animals). No difference in carbachol responsiveness could be detected between the different dietary groups under non-sensitized conditions. The results are discussed in view of the current concepts for bronchial hyperreactivity, especially in relation to eicosanoid involvement.

Topics: alpha-Linolenic Acid; Animals; Bronchial Diseases; Carbachol; Dietary Fats; Fatty Acids, Unsaturated; Guinea Pigs; Haemophilus influenzae; Histamine; Hypersensitivity; Linoleic Acid; Linoleic Acids; Linolenic Acids; Male; Muscle Contraction; Ovalbumin; Receptors, Cholinergic; Receptors, Histamine; Trachea

We examined the effects of glucocorticosteroids (GCS) on antigen-induced bronchial anaphylactic reactions (BAR) in SD rats immunized with ovalbumin (OA) and alum. The animals were treated with vehicle, budesonide (BUD), dexamethasone (DEX), or hydrocortisone (HC) at various times before intravenous (i.v.) antigen challenge. The drugs were administered either intraperitoneally (i.p.) or intratracheally (i.t.); the BAR was elicited by a low or by a high challenge dose of antigen. A BAR elicited by a low challenge dose of antigen was reduced in a dose-dependent way by all GCS after i.p. administration; at 1 mg/kg, BUD and DEX significantly reduced BAR and at 50 mg/kg all three of the examined compounds inhibited the BAR by 50% or more. For BUD, maximum effect was recorded when it was given 12 h before test. There was only a slight variation in the inhibitory effects of the GCS with immunization conditions of test animals. I.t. instillation of the drugs did not markedly increase their inhibitory capacity as compared to i.p. administration. BAR elicited by a high antigen dose was at best marginally affected by the GCS when given either i.p. or i.t. Thus, antigen-induced airway reactivity in rats can be reduced by GCS treatment provided that this is performed sufficiently long before the test and that the challenge dose of antigen is not too high.

Topics: Acetophenones; Administration, Topical; Alum Compounds; Aluminum; Anaphylaxis; Animals; Antigens; Bronchial Diseases; Budesonide; Cromolyn Sodium; Dexamethasone; Dose-Response Relationship, Drug; Drug Interactions; Glucocorticoids; Immunization; Injections, Intraperitoneal; Injections, Intravenous; Male; Ovalbumin; Pregnenediones; Quinacrine; Rats; Rats, Inbred Strains; Respiration; Sulfates; Time Factors

Rats (BN X Wistar) and mice (CBA/Ca) were immunized by exposure in 10-day periods to an aerosol of ovalbumin (OA). In rats this immunization resulted in IgE antibodies detectable at very low levels in bronchial washings, whereas IgG, IgA and IgM antibodies were recorded both in serum and in bronchial washings. In mice, exposure to aerosolized antigen resulted in specific IgE and IgG antibodies in serum. The levels of IgM antibodies were low and no IgA antibodies could be recorded with the enzyme-linked immunosorbent assay (ELISA). Histological examination of lung tissue from immunized rats and mice revealed increased numbers of cells with characteristics of both immature and mature mast cells. In addition, in the rats these cells were more closely located to the bronchi in immunized than in control animals. In the latter animals the mast cells were located around the blood vessels. Immature mast cells were located in the bronchiole-associated lymphatic tissue (BALT) which showed a marked proliferation in immunized animals. The findings indicate that sensitization via the airways provides possibilities to develop a model in rodents for studies of IgE-mediated allergy in the lung.

Topics: Aerosols; Animals; Antibody Formation; Antigens; Bronchi; Bronchial Diseases; Disease Models, Animal; Female; Granuloma; Immunoglobulin E; Immunoglobulins; Lung; Male; Mast Cells; Mice; Mice, Inbred CBA; Ovalbumin; Rats; Rats, Inbred Strains

Brown-Norway rats, sensitized with trinitrophenyl (TNP) haptenized ovalbumin and AIPO4 as adjuvant 12 days before, were challenged with trinitrophenyl haptenized bovine serum albumin intravenously, while lung function (Vt, V, Ppl, Fres, Cdyn, and Rl) and cardiovascular function (BP and Fheart) were measured continuously. This resulted in a highly reproducible, plasma IgE-antiTNP related, immediate anaphylactic response characterized by a short-lasting (8-10 min) bronchoconstriction, together with a long-lasting fall in blood pressure. All rats died in shock within 21-150 min. This method is simple and appeared to be highly reproducible and therefore suitable to screen or study antiallergic drugs in vivo.

Topics: Anaphylaxis; Animals; Bronchial Diseases; Cardiovascular Diseases; Disease Models, Animal; Female; Haptens; Immunoglobulin E; Male; Ovalbumin; Rats; Rats, Inbred BN; Serum Albumin, Bovine; Trinitrobenzenes