ovalbumin and Alveolitis--Extrinsic-Allergic

Topics: Alveolitis, Extrinsic Allergic; Animals; Asthma; Dexamethasone; Inflammation; Macrophages; Male; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phenotype; Pneumonia; Rats; Rats, Wistar

The role and origin of alveolar macrophages (AMs) in asthma are incompletely defined. We sought to clarify these issues in the context of acute allergic lung inflammation using house dust mite and OVA murine models. Use of liposomal clodronate to deplete resident AMs (rAMs) resulted in increased levels of inflammatory cytokines and eosinophil numbers in lavage fluid and augmented the histopathologic evidence of lung inflammation, suggesting a suppressive role for rAMs. Lung digests of asthmatic mice revealed an increased percentage of Ly6C(high)/CD11b(pos) inflammatory monocytes. Clodronate depletion of circulating monocytes, by contrast, resulted in an attenuation of allergic inflammation. A CD45.1/CD45.2 chimera model demonstrated that recruitment at least partially contributes to the AM pool in irradiated nonasthmatic mice, but its contribution was no greater in asthma. Ki-67 staining of AMs supported a role for local proliferation, which was increased in asthma. Our data demonstrate that rAMs dampen, whereas circulating monocytes promote, early events in allergic lung inflammation. Moreover, maintenance of the AM pool in the early stages of asthmatic inflammation depends on local proliferation, but not recruitment.

Topics: Allergens; Alveolitis, Extrinsic Allergic; Animals; Antigens, Ly; Asthma; Bronchoalveolar Lavage Fluid; CD11b Antigen; Cell Proliferation; Clodronic Acid; Cytokines; Disease Models, Animal; Eosinophils; Inflammation; Leukocyte Common Antigens; Lung; Macrophages, Alveolar; Mice; Mice, Inbred C57BL; Monocytes; Ovalbumin; Pneumonia; Pyroglyphidae

Schistosoma mansoni infection has been associated with protection against allergies. The mechanisms underlying this association may involve regulatory cells and cytokines. We evaluated the immune response induced by the S. mansoni antigens Sm22.6, PIII and Sm29 in a murine model of ovalbumin (OVA)-induced airway inflammation. BALB/c mice were sensitized with subcutaneously injected OVA-alum and challenged with aerolized OVA. Mice were given three doses of the different S. mansoni antigens. Lung histopathology, cellularity of bronchoalveolar lavage (BAL) and eosinophil peroxidase activity in lung were evaluated. Immunoglobulin (Ig)E levels in serum and cytokines in BAL were also measured. Additionally, we evaluated the frequency of CD4+forkhead box P3 (FoxP3)+ T cells in cultures stimulated with OVA and the expression of interleukin (IL)-10 by these cells. The number of total cells and eosinophils in BAL and the levels of OVA-specific IgE were reduced in the immunized mice. Also, the levels of IL-4 and IL-5 in the BAL of mice immunized with PIII and Sm22.6 were decreased, while the levels of IL-10 were higher in mice immunized with Sm22.6 compared to the non-immunized mice. The frequency of CD4+FoxP3+ T cells was higher in the groups of mice who received Sm22.6, Sm29 and PIII, being the expression of IL-10 by these cells only higher in mice immunized with Sm22.6. We concluded that the S. mansoni antigens used in this study are able to down-modulate allergic inflammatory mediators in a murine model of airway inflammation and that the CD4+FoxP3+ T cells, even in the absence of IL-10 expression, might play an important role in this process.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antigens, Helminth; Asthma; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Female; Forkhead Transcription Factors; Immunization; Interleukins; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia; Schistosoma mansoni; T-Lymphocyte Subsets

The role of CD8(+) T cells in oral tolerance remains unclear. To address this, we developed a model to induce CD8(+) Tregs by feeding the major histocompatibility complex class I immunodominant epitope of OVA, OVA((257-264)). OVA((257-264)) feeding induced tolerance similar to that observed in OVA protein-fed mice, capable of suppressing the production of Th1 and Th17 cytokines and inhibiting a Th1-driven delayed-type hypersensitivity response following immunization with whole OVA (wOVA) protein. OVA((257-264)) peptide-induced suppression could be transferred to naive mice with CD8(+) cells, but not CD8-depleted cells, isolated from mesenteric lymph nodes of peptide-fed mice. Interestingly, while capable of inhibiting Th1 and Th17 responses, OVA((257-264)) feeding could not suppress any feature of a Th2 inflammatory response, though OVA protein feeding could, suggesting that these cells function through a different mechanism than their CD4(+) counterparts generated in response to feeding with wOVA. Thus, CD8(+) T cells are functionally capable of mediating tolerance to Th1 and Th17 responses.

Topics: Administration, Intranasal; Administration, Oral; Adoptive Transfer; Alveolitis, Extrinsic Allergic; Animals; Antigens; CD8-Positive T-Lymphocytes; Desensitization, Immunologic; Ear, External; Edema; Immune Tolerance; Immunization; Immunodominant Epitopes; Injections, Intradermal; Lymphocyte Cooperation; Mice; Mice, Inbred C57BL; Models, Immunological; Ovalbumin; Peptide Fragments; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

The effects of single or repeated amphetamine (AMPH) treatment and those of AMPH withdrawals on immune-mediated lung inflammatory response were studied in rats. Two experiments were done. In the first, rats egg-albumin (OVA) sensitized were singularly or repeatedly (21 days, once daily) treated with AMPH (1.0 mg/kg) or with a similar number and volume of 0.9% NaCl. The OVA aerosol challenge was performed 12 h after the single or last repeated AMPH treatment and also 72 and 120 h after AMPH withdrawal. In the second experiment, the effects of reserpine (1.0 mg/kg/day for 5 consecutive days) on single AMPH actions on lung allergic response of rats were analyzed. Single and repeated AMPH treatment induced opposite actions on Bronchoalveolar lavage fluid (BAL) cellularity of allergic rats: single treatment decreased and repeated treatment increased the total number of cells as well as those of macrophages, neutrophils and eosinophils. Our data also showed that single but not repeated AMPH treatment decreased the number of neutrophils, monocytes and lymphocytes in the peripheral blood, and increased the total number of bone marrow cells in rats sensitized and challenged with OVA. Furthermore, it was shown that reserpine treatment precluded the effects of single AMPH treatment on cellular migration to the lung of OVA-sensitized and challenged rats. It was concluded that AMPH effects on lung inflammatory response and cell recruitment to the lung in allergic rats rely at least partially on corticosterone serum levels. The possible involvement of vesicular monoamine transporter type 2 (VMAT2) with these observed effects was discussed.

Topics: Alveolitis, Extrinsic Allergic; Amphetamine; Animals; Antipsychotic Agents; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Corticosterone; Leukocyte Count; Lung; Male; Ovalbumin; Rats; Rats, Wistar; Reserpine; Substance Withdrawal Syndrome; Vesicular Monoamine Transport Proteins

Asthma is a chronic inflammatory disease of the airways associated with a Th2 immune response. Despite their side effects, corticosteroids are the most used and effective drugs for treatment of asthma. In this work we investigated the efficacy of lupeol, a triterpenoid isolated from Lonchocarpus araripensis [corrected] Benth. (Fabaceae), in the treatment of bronchial asthma in BALB/c mice immunized with ovalbumin. Administration of lupeol caused the reduction of cellularity and eosinophils in the bronchoalveolar lavage fluid. Treatment with lupeol also reduced the production of mucus and overall inflammation in the lung. Levels of Type II cytokines IL-4, IL-5 and IL-13 were significantly reduced in mice treated with lupeol, an effect that was similar to that observed in dexamethasone-treated mice. In contrast, IgE production was not significantly altered after treatment with lupeol. In conclusion, our results demonstrate that lupeol attenuates the alterations' characteristics of allergic airway inflammation. The investigation of the mechanisms of action of this molecule may contribute for the development of new drugs for the treatment of asthma.

Topics: Alveolitis, Extrinsic Allergic; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antibodies; Bronchoalveolar Lavage Fluid; Cytokines; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pentacyclic Triterpenes; T-Lymphocytes, Helper-Inducer; Triterpenes

Hydroquinone (HQ) is naturally found in the diet, drugs, as an environmental contaminant and endogenously generated after benzene exposure. Considering that HQ alters the immune system and its several source of exposures in the environment, we hypothesized that prolonged exposure of HQ could affect the course of an immune-mediated inflammatory response. For this purpose, male Wistar rats were intraperitoneally exposed to vehicle or HQ once a day, for 22 days with a 2-day interval every 5 days. On day 10 after exposure with vehicle or HQ, animals were ovalbumin (OA)-sensitized and OA-aerosolized challenged on day 23. HQ exposure did not alter the number of circulating leukocytes but impaired allergic inflammation, evidenced by lower number of leukocytes in the bronchoalveolar lavage fluid 24h after OA-challenge. Reduced force contraction of ex vivo tracheal segments upon OA-challenge and impaired mesentery mast cell degranulation after in situ OA-challenge were also detected in tissues from HQ exposed animals. The OA-specificity on the decreased responses was corroborated by normal trachea contraction and mast cell degranulation in response to compound 48/80. In fact, lower levels of circulating OA-anaphylactic antibodies were found in HQ exposed rats, as assessed by passive cutaneous anaphylaxis assay. The reduced level of OA-anaphylactic antibody was not dependent on lower number or proliferation of lymphocytes. Nevertheless, lower expression of the co-stimulatory molecules CD6 and CD45R on OA-activated lymphocytes from HQ exposed rats indicate the interference of HQ exposure with signaling of the humoral response during allergic inflammation. Together, these data indicate specific effects of HQ exposure manifested during an immune host defense.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Bronchoalveolar Lavage Fluid; Cell Degranulation; Cell Proliferation; Environmental Pollutants; Flow Cytometry; Hydroquinones; Leukocyte Common Antigens; Leukocyte Count; Lymphocytes; Male; Mast Cells; Muscle Contraction; Muscle, Smooth; Neutrophil Infiltration; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Wistar; Spleen; Trachea

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

We have reported that Ganoderma tsugae supplementation alleviates bronchoalveolar inflammation in an airway sensitization and challenge model with female BALB/c mice. However, the effects of G. tsugae supplementation in vivo on serum antibody levels, splenocyte and peritoneal microphage immune responses have not yet been determined. In this study, serum antibody levels, cytokines and splenocyte chemical mediators and peritoneal macrophage cultures from ovalbumin (OVA)-sensitized and -challenged mice were examined after continuously consuming G. tsugae supplementation diets for 5 weeks. The results showed that OVA sensitization and challenge significantly (P<0.05) decreased the spontaneous production of IL-2 (Th1) cytokine, but significantly (P<0.05) increased spontaneous and OVA-stimulated IL-4 (Th2) production in splenocyte cultures from experimental mice. OVA administration significantly decreased both spontaneous and LPS/IFN-gamma-stimulated IL-1beta and IL-6 levels in peritoneal macrophage cultures from experimental mice. However, dietary supplementation with G. tsugae significantly increased spontaneous IL-2 level, but slightly decreased spontaneous IL-4 level in cultured splenocyte supernatants in the experimental groups. G. tsugae supplementation enhanced pro-inflammatory cytokines IL-1beta and IL-6 production in cultured peritoneal macrophages. However, the nitric oxide level from cultured peritoneal macrophages and serum OVA-specific IgE and IgG(2a) antibody levels was not significantly affected. These results suggest that OVA sensitization and challenge induced a Th2-skewed splenocyte response and decreased peritoneal macrophage cytokine secretion. G. tsugae supplementation in vivo modulated the Th1/Th2 balance and enhanced macrophage immune responses. However, the supplementation diet could not fully reverse the Th2-skewed responses to level of Th1-skewed responses.

Topics: Alveolitis, Extrinsic Allergic; Animals; Dietary Supplements; Disease Models, Animal; Drugs, Chinese Herbal; Female; Fruiting Bodies, Fungal; Ganoderma; Immunoglobulins; Interferon-gamma; Interleukins; Lipopolysaccharides; Macrophages, Peritoneal; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Mycelium; Ovalbumin; Phytotherapy; Spleen; Th1 Cells; Th2 Cells

T-helper (Th)1 cells have a pivotal role in the pathogenesis of hypersensitivity pneumonitis. Continued low-level exposure to the antigens may induce chronic hypersensitivity pneumonitis with lung fibrosis. Although such exposure may activate Th1 cells in the lung, it is not known whether activation of Th1 cells per se can lead to pulmonary fibrosis. To determine this, the lung pathology induced by Th1 clones was investigated. Mice (BALB/c) were injected intraperitoneally with Th1 clones 1-4 times. Each injection was performed 4 days apart and was followed by repeated exposure to aerosolised ovalbumin (OVA) once a day for 5 days. The number of macrophages and lymphocytes in bronchoalveolar lavage fluids (BALF) increased as the number of Th1 transfers increased. The number of neutrophils also increased but peaked in the second transfer and then decreased following further transfers. Increased cell infiltration, thickness of alveolar walls and number of type II cells in the lung occurred. However, histological findings showed no evidence of fibrosis and hydroxyproline levels did not increase. Findings of histology and BALF were ameliorated 2 weeks after the discontinuation of OVA exposure, indicating the reversibility of the Th1-induced pathology. In conclusion, adoptive transfer of T-helper 1 cells results in reversible alveolitis but does not lead to pulmonary fibrosis.

Topics: Adoptive Transfer; Alveolitis, Extrinsic Allergic; Animals; Female; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Fibrosis; T-Lymphocytes, Helper-Inducer

We recently demonstrated a codominant role of C5aR and FcgammaRIII in the initiation of IgG immune complex-mediated inflammation in mice. In this study, we investigated the relative contribution of FcgammaRIII in the generation of several cytokines during experimental hypersensitivity pneumonitis/alveolitis in vivo. Induction of immune complex-alveolitis in C57BL/6 mice resulted in strong accumulation of neutrophils into the lung and enhanced chemotactic activity within bronchoalveolar lavage fluid accompanied by an increased production of the proinflammatory cytokines TNF-alpha and IL-1beta as well as the ELR-CXC chemokines macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant (KC). FcgammaRIII-deficient C57BL/6 mice (FcgammaRIII(-/-)) showed a marked reduction of the inflammatory response due to decreased production of TNF-alpha, IL-1beta, and MIP-2. Results obtained in C57BL/6 mice either lacking the TNF-alpha class I receptor (TNF-alphaRI(-/-)) or treated with neutralizing anti-TNF-alpha mAb demonstrated an essential contribution of TNF-alpha for mediating IL-1beta release, neutrophil influx, and hemorrhage. Surprisingly, MIP-2 and KC chemokine levels remained largely unaffected in TNF-alphaRI(-/-) mice or after functional inhibition of TNF-alpha. These data suggest that in immune complex alveolitis, the activation of FcgammaRIII may induce divergent downstream effector pathways with TNF-alpha acting independently of CXC chemokines to trigger the inflammatory response in C57BL/6 mice.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antigen-Antibody Complex; Bronchoalveolar Lavage Fluid; Cell Movement; Chemokine CXCL2; Chemokines; Chemokines, CXC; Chemotaxis, Leukocyte; Cytokines; Immune Complex Diseases; Immunoglobulin G; Injections, Intravenous; Interleukin-1; Intubation, Intratracheal; Kinetics; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Ovalbumin; Receptors, IgG; Tumor Necrosis Factor-alpha

Airway inflammation, hyperreactivity, increased number of goblet cells, and mucus overproduction characterize asthma. Respiratory challenge with ovalbumin (OVA) of sensitized mice has been shown by several laboratories to cause pulmonary pathology similar to that observed in human allergic asthma. Recently, interleukin (IL)-13 has been shown to be a central mediator in this process. Because the airways of healthy mice have few, if any, mucus-producing cells, an increase in the number of these cells likely reflects induction of mucin-gene expression. The purpose of this study was to identify mucin genes induced as a result of airway goblet-cell metaplasia (GCM) in mice sensitized and challenged with OVA or in mice treated with IL-13 alone. BALB/c mice were sensitized by intraperitoneal injection (Days 0, 4, 7, 11, and 14) and intranasal instillation (Day 14) of 100 microg of OVA in saline, and then challenged by intranasal instillation (Days 25, 26, and 27) of the same. IL-13-treated mice received 5 microg of IL-13 by intranasal instillation on three consecutive days. Control mice were given saline alone. All mice were studied 24 h after the last challenge. Histologic analysis of the lungs revealed both a striking peribronchial and perivascular lymphocytic and eosinophilic inflammation and airway GCM in OVA-treated mice, and also airway GCM without inflammation in IL-13-treated mice. Northern blot analysis of lung RNA demonstrated (1) expression of Muc-5/5ac messenger RNA (mRNA) in OVA-treated and IL-13-treated mice, but not in control mice; (2) expression of Muc-1 mRNA at comparable levels in all mice regardless of treatment; and (3) no expression of Muc-2 or Muc-3 mRNA in control or treated mice. Western blot analysis demonstrated the expression of Muc-5/5ac protein (both apomucin and glycosylated mucin) in lung lysates of OVA-treated (but not control) mice, and also the expression of Muc-5/5ac mucins in the bronchoalveolar lavage fluid of OVA-treated and IL-13-treated mice. These findings demonstrate that airway GCM is associated with the induction of pulmonary expression of Muc-5/5ac mRNA and mucin in murine models of allergic asthma.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antibodies; Biomarkers; Bronchoalveolar Lavage Fluid; Gene Expression; Goblet Cells; Interleukin-13; Lung; Male; Metaplasia; Mice; Mice, Inbred BALB C; Mucin 5AC; Mucin-5B; Mucins; Ovalbumin; RNA, Messenger

Airway wall remodeling in response to inflammation might alter load on airway smooth muscle and/or change airway wall stability. We therefore determined airway wall compliance and closing pressures in an animal model. Weanling pigs were sensitized to ovalbumin (OVA; ip and sc, n = 6) and were subsequently challenged three times with OVA aerosol. Control pigs received 0.9% NaCl (n = 4) in place of OVA aerosol. Bronchoconstriction in vivo was assessed from lung resistance and dynamic compliance. Semistatic airway compliance was recorded ex vivo in isolated segments of bronchus, after the final OVA aerosol or 0.9% NaCl challenge. Internally or externally applied pressure needed to close bronchial segments was determined in the absence or presence of carbachol (1 microM). Sensitized pig lungs exhibited immediate bronchoconstriction to OVA aerosol and also peribronchial accumulations of monocytes and granulocytes. Compliance was reduced in sensitized bronchi in vitro (P < 0.01), and closing pressures were increased (P < 0.05). In the presence of carbachol, closing pressures of control and sensitized bronchi were not different. We conclude that sensitization and/or inflammation increases airway load and airway stability.

Topics: Air Pressure; Allergens; Alveolitis, Extrinsic Allergic; Animals; Bronchi; Carbachol; Female; Lung Compliance; Lung Volume Measurements; Muscarinic Agonists; Muscle Tonus; Ovalbumin; Swine

In the present study, we investigated whether allergen immunotherapy is effective in a murine model with immunologic and pathophysiologic features reminiscent of allergic asthma. Ovalbumin-sensitized mice received increasing (1 microgram to 1 mg) subcutaneous doses of ovalbumin twice a week for 8 wk according to a semirush immunotherapy protocol as used in allergic patients. During immunotherapy, an initial rise in serum levels of ovalbumin-specific antibodies (immunoglobulin [Ig]G1, IgE, IgG2a) occurred, after which IgE levels decreased sharply concomitant with an increase in IgG2a levels. The increase in IgG2a levels, with the decline in IgE levels, suggests that during immunotherapy interferon-gamma production is increased or interleukin (IL)-4 production is decreased. After immunotherapy, inhalation challenge of the mice with ovalbumin revealed almost complete inhibition (98%, P < 0.01) of eosinophil infiltration into bronchoalveolar lavage and airway hyperresponsiveness (100% at 320 microgram/kg methacholine, P < 0.05) compared with sham-treated animals. In addition, IL-4 production of thoracic lymph node cells stimulated with ovalbumin in vitro was largely reduced (60%, P < 0.05) after immunotherapy. Thus, effective immunotherapy in this animal model appears to be due to modulation of antigen-specific T cells. Similar effects on airway symptoms and IL-4 production can be obtained within 1 wk by three injections of the highest dose of ovalbumin (1 mg). This animal model will be used as a preclinical model to improve allergen immunotherapy and to gain more insight into the mechanisms involved.

Topics: Alveolitis, Extrinsic Allergic; Animals; Asthma; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Desensitization, Immunologic; Disease Models, Animal; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-4; Lymph Nodes; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Eosinophilia; Specific Pathogen-Free Organisms; T-Lymphocytes

Cultured CD4+ cells are responsible for transfer of adoptive murine experimental hypersensitivity pneumonitis (EHP) (ARRD 1992; 146:1582-8). To characterize interactions that occur in vitro that result in cells able to transfer EHP, we added either antibody to IFN-gamma, antibody to IL-2, or 30 or 300 micrograms/ml IFN-gamma at the onset of 72-hour culture of C3H/HeJ spleen cells from either M. faeni or ovalbumin (control) sensitized donors with 30 micrograms/ml Micropolyspora faeni. We determined the phenotype of cells after culture and the amount of IL-2 or IFN-gamma in the culture supernatants, transferred cells to naive recipients, challenged the recipients intratracheally with M. faeni, and determined the extent of pulmonary inflammatory changes 4 days thereafter. Substantial amounts of IL-2 and IFN-gamma were detected in supernatants of cultures from M. faeni-sensitized animals, and lesser amounts were detected in culture supernatants from ovalbumin-sensitized donors. Treatment of cultures of M. faeni-sensitized cells with antibody to IL-2 or IFN-gamma blocked or reduced measurable IL-2 or IFN-gamma for the duration of culture. Treatment with IFN-gamma blocked increased levels of IL-2 at 48 and 72 hours of culture. Cultured M. faeni-sensitized cells adoptively transfer EHP. Cells from cultures depleted of either IL-2 or IFN-gamma or supplemented with IFN-gamma could transfer EHP equally well. We conclude that in vitro maturation of cells capable of adoptive EHP is not dependent on soluble IL-2 or IFN-gamma and is not altered by exogenous IFN-gamma.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antibodies, Monoclonal; Antigens, Bacterial; Cytokines; Immunophenotyping; Immunotherapy, Adoptive; Interferon-gamma; Interleukin-2; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Micromonosporaceae; Ovalbumin; Spleen; Th1 Cells; Th2 Cells

BN50739 is a new and potent PAF antagonist. In this experiment, we have further confirmed that it is a selective PAF antagonist by means of rabbit platelet aggregating test. And also, when BN50739 was applied, in the concentration of 10 uM, to the isolated lung of guinea pig sensitized by ovalbumin, we observed that the amount of histamine and thromboxane A2 released was a little less (about 19.0% and 19.2%, respectively) than that of controls (P < 0.05). Whereas, the ratio of PGI2/TxA2 was increased by 17.6% comparing to that in control group (P < 0.05). But the level of LTC4 remained unchanged regardless of the presence of BN50739. We conclude that PAF may play a role in allergic reaction.

Topics: Alveolitis, Extrinsic Allergic; Animals; Azepines; Female; Guinea Pigs; Histamine; Lung; Male; Ovalbumin; Platelet Activating Factor; Prostaglandins F; SRS-A; Thromboxane B2; Triazoles

To characterize the cells responsible for transfer of adoptive murine experimental hypersensitivity pneumonitis (EHP), we depleted Micropolyspora faeni (M. faeni)-sensitized C3H/HeJ spleen cell (SC) cultures of CD3+, CD4+, or CD8+ cells before administration to recipients. We determined the length of time sensitization persists, the ability of cultured lung-associated lymph node (LALN) cells to transfer EHP, and the ability of cultured SC from animals subjected to two, four, or eight weekly intratracheal challenges of M. faeni to transfer EHP. The extent of pulmonary inflammatory response after challenge with intratracheal M. faeni was used to determine adoptive transfer. Depletion reduced the proportion of CD3+ cells from 21 to 1%, CD4+ cells from 15 to 3%, and CD8+ cells from 7 to 1% in the cultured SC population. The proportion of B cells exhibited reciprocal changes. Cultured SC could transfer EHP. Depletion of CD3+ and CD4+, but not CD8+ cells, ablated or diminished the capacity to transfer EHP. Sensitized cells persisted in recipients for at least 8 wk. Cultured LALN cells could transfer EHP. Recipients of cultured SC from 4- and 8-wk donors exhibited less extensive pulmonary abnormalities than recipients of cultured SC from 2-wk donors. The proportion of CD3+, CD4+, CD8+, B cells, and macrophages was the same in cultured cells from 2-, 4-, and 8-wk donors. We conclude that the active cells in SC cultures are CD3+, CD4+, and CD8- T cells, and there are differences in the ability of cultured cells to adoptively transfer EHP that are dependent on the nature of the donor but not on the phenotype of the cell population.

Topics: Alveolitis, Extrinsic Allergic; Animals; CD3 Complex; CD4 Antigens; Cells, Cultured; Disease Susceptibility; Immunization; Immunophenotyping; Immunotherapy, Adoptive; Lung; Lymphocyte Depletion; Male; Mice; Mice, Inbred C3H; Micromonosporaceae; Ovalbumin; T-Lymphocytes

Rabbit models of chronic experimental hypersensitivity pneumonitis and desensitization were used to evaluate the effects of systemic cyclosporine. When administered 12 to 18 h before each inhalational challenge with aerosolized antigen and the adjuvant muramyl dipeptide, cyclosporine suppressed the development of disease as well as the anamnestic antibody response, particularly in bronchoalveolar lavage fluids. When administered at the time of sensitization only, cyclosporine suppressed the primary antibody response but not the anamnestic antibody response or the disease. Antigen- and mitogen-induced blastogenesis was inhibited by cyclosporine in vitro, but antigen-specific blastogenesis was not abrogated by cyclosporine previously administered in vivo. These results indicate that cyclosporine caused profound immunomodulation in this model, which can be at least partially explained by transient suppressive effects on T cells, particularly the helper/inducer and delayed hypersensitivity subset(s).

Topics: Alveolitis, Extrinsic Allergic; Animals; Antibody Formation; Chronic Disease; Cyclosporins; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Female; Hypersensitivity, Delayed; Immunoglobulin G; Lymphocytes; Lymphokines; Ovalbumin; Rabbits; Therapeutic Irrigation

Fluorescein isothiocyanate (FITC) conjugated to protein carriers was used to explore carrier dependence in an established rabbit model of acute hypersensitivity pneumonitis (HSP). Rabbits were immunized via toepads with either FITC-ovalbumin (OA) or FITC-human gamma-globulin (HGG) in complete Freund's adjuvant, and were aerosol challenged with homologous or heterologous conjugates 30 days later. Only those rabbits challenged with the homologous carrier developed acute HSP, despite the presence of comparable levels of anti-FITC antibodies in the sera of all groups. These findings indicate a strict carrier dependence in the pathogenesis of HSP in this model and provide further evidence that the mechanism of inflammation depends upon a cellular immune response.

Topics: Acute Disease; Alveolitis, Extrinsic Allergic; Animals; Antibody Formation; Carrier Proteins; Disease Models, Animal; Fluorescein-5-isothiocyanate; Fluoresceins; Fluorescent Antibody Technique; Haptens; Humans; Lung; Ovalbumin; Rabbits; Thiocyanates

An established rabbit model of acute hypersensitivity pneumonitis was used to evaluate adjuvant properties of synthetic muramyl dipeptide (MDP), the minimal adjuvant-active structure of mycobacteria. The authors studied MDP as a substitute for mycobacteria in immunization and as adjuvant during repeated inhalation of antigen (ovalbumin). They found that MDP could substitute successfully for mycobacteria in sensitizing animals for acute alveolitis following subsequent inhalation of a combination of ovalbumin and MDP aerosol for 4 to 14 weeks resulted in the development of chronic granulomatous pneumonitis, characterized by alveolar wall thickening, granulomas, and infiltrations with lymphocytes and macrophages. In addition, MDP boosted systemic and local IgG and IgA antigen-specific antibodies. Inhaled MDP, itself neither antigenic nor mitogenic, acted therefore as adjuvant for continued immunologic inflammatory effector mechanisms in the rabbit lung, which are ordinarily suppressed when antigen alone is inhaled. Possible mechanisms include stimulation of effector T cells and macrophages or the failure of suppressive mechanisms, with or without participation of immune complexes. This is the first successful model of chronic granulomatous alveolitis produced by inhalation of soluble materials. Further exploration of adjuvant mechanisms in this system should help clarify the pathogenesis of immunologic lung diseases in man.

Topics: Acetylmuramyl-Alanyl-Isoglutamine; Alveolitis, Extrinsic Allergic; Animals; Bacterial Vaccines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Female; Glycopeptides; Immunization; Lymphocytes; Male; Mycobacterium; Ovalbumin; Pulmonary Alveoli; Rabbits; Respiration

Topics: Alveolitis, Extrinsic Allergic; Animals; Buffaloes; Cattle; Female; Food Hypersensitivity; Humans; Infant; Milk; Ovalbumin; Precipitins

We evaluated local and systemic humoral responses to inhalational challenges with ovalbumin (OA) by measuring IgA and IgG isotypic antibodies to OA in serum and bronchoalveolar wash fluids (BAW), and by quantitating cells containing IgA, IgG, and anti-OA in lung, lymph nodes, spleen, and gut. Rabbit models of acute hypersensitivity pneumonitis and chronically-challenged "desensitized" animals were studied along with appropriate control animals. Systemic (via a toe pad) immunization or acute aerosol challenge with OA resulted in only trace amounts of IgG anti-OA antibodies in BAW and no apparent anti-OA-containing cells in the lung itself. Acute aerosol challenge of systemically immunized rabbits caused alveolitis, increased IgG anti-OA-containing cells in mediastinal and popliteal lymph nodes, and increased IgG anti-OA in BAW attributable to transudation from serum. Thrice-weekly inhalational challenge with aerosolized OA resulted in waning alveolitis, elevated concentrations of IgA and IgG anti-OA in BAW and serum, and increased concentrations of IgG and IgA anti-OA cells in the lung, but not in other tissues, including the gut. We conclude that these experiments have implicated IgG as well as IgA antibodies in local humoral responses to inhaled antigen, have not substantiated the notion of a common mucosal immune system involving lung and gut, have failed to demonstrate humoral tolerogenesis after inhalation of antigen, and have shown an effect of systemic priming on subsequent pulmonary immune responses in the models examined.

Topics: Alveolitis, Extrinsic Allergic; Animals; Bronchi; Immunoglobulin A; Immunoglobulin G; Ovalbumin; Pulmonary Alveoli; Rabbits; Time Factors

Rabbits that had been prepared to develop acute alveolitis after aerosol challenge with simple protein antigens did not develop chronic alveolitis but rather gradually recovered despite continued challenge. Immunologic accompaniments of waning disease were compared in this model to those associated with intravenous injections of antigen causing "desensitization." We also studied the effects of aerosol challenge prior to systemic immunization, antigen specificity, and the duration of desensitization by aerosolized and intravenous antigen. We found that repeated aerosol or intravenous challenges produced antigen-specific desensitization in this model, and the effect lasted several weeks. Prior exposure to aerosolized antigen was not protective. Neither aerosol nor intravenous desensitization maneuvers abrogated antigen-specific lymphocyte blastogenesis, although an early transient fall did occur. Humoral responses were boosted. These findings suggest that chronic alveolitis is prevented in this model by specific desensitization, without the induction of true tolerance or of nonspecific anergy. Such immunoregulation may result from development of antigen-specific blockade or blocking factors (e.g., lymphokines), antigen-antibody complexes, or suppressor cells affecting specific effector cells. Evaluation of these mechanisms may have implications for diagnosis and prognosis in human hypersensitivity pneumonitis.

Topics: Acute Disease; Administration, Intranasal; Alveolitis, Extrinsic Allergic; Animals; Antigens; Cattle; Desensitization, Immunologic; Disease Models, Animal; Epitopes; Female; gamma-Globulins; Immunoglobulin A; Injections, Intravenous; Lymphocyte Activation; Male; Ovalbumin; Rabbits; Time Factors

The earliest lesion in hypersensitivity penumonitis is an acute inflammatory alveolitis characterized by parenchymal hemorrhage and accumulations of polymorphonuclear leukocytes within the lung. In many instances, this initial lesion is replaced by a more chronic alveolitis, with development of mononuclear cell interstitial infiltrate, granuloma formation, and interstitial fibrosis. To help define the mechanisms by which the early polymorphonuclear leukocyte alveolitis of acute hypersensitivity pneumonitis evolves into a chronic mononuclear-cell process, an animal model of the disease was developed using guinea pigs sensitized by footpad injeection with either ovalbumin (OVA) in complete Freund's adjuvant (CFA), CFA alone, or phosphate-buffered saline. Ten days after sensitization, the animals were challenged by intratracheal injection of either particulate OVA, particulate human serum albumin, or phosphate-buffered saline alone, and their lungs were evaluated sequentially for changes in histologic appearance and lymphocyte subpopulations. After challenge, only animals sensitized with CFA plus OVA and challenged with particulate OVA developed pulmonary lesions consistent with acute hypersensitivity pneumonitis. Within 4 h after challenge, these animals developed an acute hemorrhagic alveolitis that persisted for more than 24 h. By 48 to 96 h, the alveolitis evolved into a predominantly mononuclear-cell infiltrate. This change in the histologic appearance of the lungs in these animals was preceded by a rapid increase in the proportions of T-lymphocytes within the lungs, noted by 24 h after intratracheal challenge with specific antigen. Before intratracheal challenge with antigen, lung lymphocytes from only the group of animals immunized with CFA plus OVA were capable of proliferating on exposure to OVA in vitro. In the same group, lymphocytes recovered from the lung after intratracheal particulate OVA demonstrated blast transformation in vivo, a phenomenon not found in any other group. These studies suggest that the alveolitis of acute hypersensitivity pneumonitis is rapidly associated with changes in populations of immune effector cells before development of the mononuclear cell alveolitis characteristic of the chronic disease.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antigen-Antibody Reactions; Disease Models, Animal; Guinea Pigs; Lung; Ovalbumin; T-Lymphocytes

In previous studies with isolated perfused rabbit lungs, we observed that human serum albumin (HSA) and ovalbumin, introduced into the isolated lungs as an aerosol, entered the pulmonary circulation antigenically intact. The "inhaled" proteins were also broken down in the lung. When lungs from animals immunized with one protein inhaled the two proteins simultaneously, absorption of intact antigen was specifically reduced, and there was a nonspecific increase in the appearance of metabolites of both proteins in the blood. In the present study, we investigated the antigen-specific and nonspecific effects of two types of hypersensitivity responses on protein absorption across the air-blood barrier of isolated rabbit lungs. In one group of lungs, an acute hypersensitivity response was induced by introducing HSA into the blood perfusing lungs from HSA-immunized rabbits. In another, the rabbits had been previously exposed to chronic HSA aerosol until their lungs exhibited a chronic immunologic inflammatory response. Lungs from both groups were insufflated simultaneously with HSA, and a nonspecific protein, ovalbumin. Lungs in which the acute anaphylactic response was induced showed no alteration in the absorption of either intact protein compared with HSA-immunized controls, but absorbed a somewhat larger quantity of breakdown products of the specific antigen. Lungs undergoing the chronic alveolar inflammation were more permeable to nonspecific protein than were noninflamed lungs. Despite the increased permeability to nonspecific protein, the absorption of antigen was blocked as effectively as in immune but noninflamed controls. In these chronically inflamed lungs, the absorption of antigen breakdown products was enhanced. The results indicate that both immunologic and inflammatory mechanisms may control the amounts of inhaled soluble proteins that reach the blood via the alveolocapillary barrier. Alterations in the absorption of inhaled proteins and their metabolites across the air-blood barrier during certain types of hypersensitivity responses may be of immunologic and pathologic significance.

Topics: Absorption; Alveolitis, Extrinsic Allergic; Anaphylaxis; Animals; Antigens; Capillaries; Female; Male; Ovalbumin; Pulmonary Alveoli; Rabbits; Serum Albumin

To study the effects of steroids on the pulmonary lesions in experimental hypersensitivity pneumonitis, rabbits were sensitized to ovalbumin (OA) by injections of OA into footpads and 3 weeks later they were subjected to two successive aerosol challenges with OA at an interval of 48 hr. Injections of hydrocortisone sodium succinate 10 mg twice daily (but not at the reduced dosage of 5 mg twice daily) or methyprenisolone acetate 5 mg twice daily beginning 30 min before the first challenge and continued to the time of killing reduced the extent and intensity of vasculitis in both the treated groups and showed less alveolar septal thickening in the hydrocortisone treated group and less alveolar consolidation in the methylprednisolone treated group compared to the pulmonary lesions in the rabbits which were sensitized and then subjected to OA aerosol challenges, but received no treatment. In view of the observation that even in a steroid sensitive species like the rabbit, extensive pulmonary changes like alveolar consolidation, septal thickening and vasculitis persisted in spite of treatment with relatively large doses of these steroids, it was felt that in human hypersensitivity pneumonitis steroids might only suppress the warning symptoms without substantially affecting the progress of the pulmonary lesions.

Topics: Alveolitis, Extrinsic Allergic; Animals; Antigens; Female; Hydrocortisone; Methylprednisolone; Ovalbumin; Rabbits