J.E. Salvaggio

Basidiospore Extracts: Evidence for Common Antigenic/Allergenic Determinants

Spore extracts, prepared from Armillariella tabescens, Pleurotus ostreatus, Coprinus quadrifidus, Amanita muscaria, Ganoderma lucidum, Psilocybe cubensis, Pisolithus tinctorius, Scleroderma sp. and Calvatia cyathiformis, were examined for antigenic/allergenic relationships by Ouchterlony and radioallergosorbent testing (RAST) inhibition, respectively. Ouchterlony, using hyperimmunized rabbit sera, demonstrated a high degree of cross-antigenicity among the extracts tested; however, some unique antigens were also present. RAST inhibition, evaluated by comparing extract concentrations which inhibited the RAST by 50% (IC-50), varied with the allergen tested. P. cubensis was the most potent inhibitor (IC-50 ranged from 0.034 mg/ml for A. tabescens RAST to 0.29 mg/ml for G. lucidum RAST). P. tinctorius was the least potent inhibitor, failing to reach IC-50 at 10 mg/ml for any basidiospore extract. Evaluation of slopes and intercepts of the dose-response lines demonstrated qualitative and quantitative differences among allergens in these extracts. These results indicate the presence of shared allergenic epitopes, and suggest that representative extract panels could be developed for future use in diagnosis and treatment of basidiospore-sensitive individuals.

Toluene diisocyanate pulmonary disease: Immunopharmacologic and mecholyl challenge studies

Selected workers exhibiting clinical sensitivity to toluene diiosocyanate (TDI) (wheezing, cough, and dyspnea upon entering a TDI-containing area) were studied for : (1) in vitro TDI-induced leukocyte histamine release; (2) determination of cyclic 3,5 adenosine monophosphate (cAMP) levels of lymphocytes exposed to TDI; (3) effect of TDI on the isoproterenol-induced increase of lymphocyte cAMP levels: and (4) acetyl-beta-methylcholine (mecholyl) inhalation challenge. TDI did not induce histamine release from leukocytes of sensitive or nonsensitive individuals, nor were lymphocyte cAMP levels affected by in vitro TDI exposure, TDI did, however, diminish in vitro stimulation of cAMP by isoproterenol. This effect, seen with cells of sensitive and nonsensitive individuals, appeared to be dose-dependent; there were no significant differences between the two groups. When challenged with mecholyl, 7 of 10 sensitive but only 1 of 10 nonsensitive individuals showed a greater than 20% decrease in FEV1. These results suggest that TDI-induced obstructive airways disorders may be associated with altered beta-adrenergic function.

Passive transfer of experimental hypersensitivity pneumonitis with lymphoid cells in the rabbit

Although precipitating antibody is associated with human hypersensitivity pneumonitis, there is evidence that cell-mediated hypersensitivity may be involved in disease pathogenesis. In this study, interstitial, and peribronchial lesions were produced by respiratory challenge of rabbits passively sensitized with ovalbumin-sensitive lymph node cells. Ovalbumin sensitivity of donor rabbits and lymph node cells was demonstrated by skin testing, migration inhibition factor (MIF) production using alveolar wash cells as migrating cells, and lymphocyte stimulation. Passive cell transfer was accomplished by intraperitoneal injection with all lymph node cells obtained from one donor transferred to one recipient. Recipients were challenged by aerosol or intratracheal injection of antigen immediately or 24 hr after passive sensitization and were killed 48 hr after challenge. Lesions in rabbits passively sensitized by lymph node cells and challenged with antigen by intratracheal inoculation consisted of focal pneumonitis with intra-alveolar edema and infiltrates of mononuclear cells in alveoli and alveoli septa. Aerosol challenge of passively sensitized animals produced similar changes, but peribronchial tissue containing macrophages and germinal centers was prominent in this group. Antiovalbumin serum recipients challenged by intratracheal injection demonstrated only mild peribronchial mononuclear cell infiltrates, without pneumonitis. Control animals receiving lymph node cells only or challenge only demonstrated no changes in lung histology.

Experimental studies of sensitization to beryllium, zirconium, and aluminum compounds in the rabbit☆

In a study designed to assess the potential sensitizing and granulomagenic capacities of selected metallic salts, rabbits were inoculated intradermally with zirconium aluminum glycinate (ZAG), sodium zirconium lactate (NZL), aluminum chlorhydrate (ACH), BeSO 4, and ovalbumin (OVA) by single and multiple injections. Animals immunized with BeSO4 and with OVA developed delayed skin reactivity as well as antigen-specific alveolar macrophage migration inhibition. Neither single nor multiple injections of ZAG or ACH resulted in clear-cut positive skin reactivity, macrophage migration inhibitory factor (MIF) production, or lymphocyte stimulation. Rabbits inoculated with multiple injections of NZL (500 microng) showed some marginally positive macrophage migration inhibition and skin reactivity. Histologically, ZAG and ACH were found to induce well-organized foreign-body granulomas after intradermal injection in both normal and inoculated rabbits. NZL and BeSO4 also induced skin granulomas, but these were less organized and distinct. Cell viability and ultrastructural studies indicated that BeSO4 was highly toxic for isolated alveolar macrophages in vitro at concentrations above 10 microng/ml, but NZL and ZAG did not exert such an effect at these dose levels. BeSO4 also depressed lymphocyte stimulation in sensitized animals which demonstrated delayed skin reactivity and macrophage migration inhibition.

Fusarium solani: Prevalence of skin reactivity and antigenic allergenic analysis

The prevalence of Fusarium solani reactivity in atopic individuals with symptoms of mold allergy was assessed with skin test and RAST. In addition, F. solani preparations were evaluated for antigenic/allergenic activity. Atopic individuals tested, 24.5% (n = 69), had positive skin reactions to a phosphate-buffered saline extract of F. solani, and these responses were statistically correlated with RAST results. Immunoelectrophoretic techniques demonstrated that this extract was antigenic in rabbits and allergenic in man. Gel filtration of F. solani extract on a Bio-Gel A 0.5 m column demonstrated three peaks of ultraviolet-absorbing material. The column eluate with the greatest RAST inhibition activity was associated with a protein peak having a molecular weight greater than 341 kilodaltons; however, all peaks demonstrated inhibitory activity. These studies suggest that extracts of F. solani contain several allergens that differ in molecular weight and charge.

Activation of the alternative complement pathway by extracts of cotton dust

Extracts of cotton dust were tested for their ability to activate the alternative complement pathway in fresh normal human serum (NHS). Alternative pathway activation was determined by a haemolytic assay utilizing glutathione‐sensitized human erythrocytes, consumption of alternative pathway components in terms of alternative pathway CH50 units and an immunoelectrophoretic assay to detect split products of activation of factor B. All assays were performed under conditions that have been shown to block the initial steps of classical pathway activation but permit activation of the alternative complement pathway. Results demonstrate that the cotton dust extracts could consume alternative complement pathway proteins in a dose‐response manner. The complement activating factor is probably endotoxin since a cotton dust extract obtained by an extraction method for endotoxin yielded the greatest activity.

Immunogenic properties of tobacco smoke

The immunogenic capacity of tobacco smoke or tobacco smoke in combination with host proteins to stimulate antibody production in rabbits was investigated. Several antisera from rabbits immunized with tobacco smoke components reacted by immunoprecipitation with tobacco smoke or leaf antigens. The serum precipitins were present only in immune sera and had electrophoretic mobility and molecular size identical to immunoglobulin. These results suggest that tobacco smoke contains components capable of stimulating an immune response in rabbits. It is possible that these components are immunogenic in man, although their role as allergens remains to be determined.

Elimination of bagassosis in Louisiana paper manufacturing plant workers.

The prevalence of bagassosis was investigated in a Louisiana paper mill, which in the past had considerable numbers of workers with the disease. Based on negative clinical histories and the low number of positive serological reactions of the workers serum with Thermoactinomyces sacchari antigen, it was concluded that bagassosis was no longer present. This was thought to be due to a different method of storage of bagasse, which retards microbial decay and reduces airborne organic dust, and to the increased awareness of plant management resulting in greater safety measures.

Evaluation of immune parameters in HIV+ subjects reporting adverse reactions to sulfamethoxazole

Trimethoprim-sulfamethoxazole (TMP-SMX) is frequently used in human immunodeficiency virus (HIV)-infected patients (HIV+) for treatment or prophylaxis of Pneumocystis carinii pneumonia (PCP). Up to 80% of those patients report adverse reactions to that drug combination. To test the hypothesis that these reactions are immunologically mediated, we quantitated specific IgG and IgE SMX-human serum albumin (HSA) antibodies and immune complexes (IC) in HIV+ patients and in HIV controls. Patients with mild HIV disease had elevated specific SMX-HSA IgG and IC levels compared with those having severe disease or with controls. Conversely, patients with severe HIV disease had statistically elevated levels of specific IgE when compared with patients having milder disease or with controls. There were no differences in either specific antibody or IC levels between patients reporting adverse reactions and those who did not. Results suggest that there are increased levels of SMX-HSA-specific antibodies in some HIV+ patients. The presence of these antibodies appears to be related to severity of disease, rather than clinically significant drug sensitivity.

Effects of asbestos fibers on alveolar macrophage-mediated lymphocyte cytostasis.

Prolonged asbestos inhalation results in pulmonary inflammation and fibrosis. Since alveolar macrophages are active in regulation of immune responses in lung and appear to be involved in the pathogenesis of asbestosis, we evaluated the effects of asbestos exposure on the ability of these cells to regulate lymphocyte function. Alveolar macrophages obtained by lung lavage from BALB/C mice were treated in vitro with either UICC amosite or chrysotile asbestos and the effects on lymphocyte cytostasis compared with those of macrophages incubated with latex beads or zymosan. Macrophages (10%) incubated either alone or with latex beads for 48 hr effectively inhibited lymphocyte mitogenesis. However, alveolar macrophages incubated with either amosite or chrysotile asbestos did not demonstrate intact cytostatic activity. Decreased viability of chrysotile asbestos-treated macrophages correlated with loss of cytostatic effects, but alveolar macrophages exposed to amosite remained viable. We conclude, therefore, that exposure of alveolar macrophages to asbestos can result in loss of their ability to down-regulate lymphocyte proliferation, a finding which may be important in the pathogenesis of asbestos-related disease.