Sb Lehrer

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.

Evaluation of Basidiomycete and Deuteromycete (Fungi Imperfecti) extracts for shared allergenic determinants

Aqueous extracts of select members of the Basidiomycetes and Deuteromycetes (Fungi Imperfecti) were evaluated for the presence of shared allergenic determinants using skin prick and radio‐allergosorbent test (RAST) inhibition. Twenty adults with perennial symptoms of rhinitis, with or without asthma, were skin‐prick tested with six species of Deuteromycetes and seven species of Basidomycetes. Positive weal‐and‐flare reactivity to Pleurotus ostreatus was associated with Alternaria alternata, Fusarium solani and Epicoccum purpurescens. Positive skin reactivity to Calvatia cyathiformis was also associated with A. alternata and F. solani. Coprinus quadrifidus was associated only with F. solani, and Psilocybe cubensis was only associated with Aspergillus fumigatus. No other skin test associations were demonstrated. For every allergen tested by RAST inhibition, significant dose‐dependent homologous inhibition was demonstrated. Although the ability of an individual heterologous extract to inhibit the direct RAST varied, inhibition was generally minimal. In the most extreme example, no heterologous allergen inhibited the A. alternata RAST. However, the Armillaria tabescens RAST was inhibited 52·6%, 38·1% and 25·1% by A. fumigatus, E. purpurescens, and Penicillium notatum, respectively, suggesting significant cross‐reactivity. These results suggest that, although shared allergenic determinants exist between select species of Basidiomycetes and Deuteromycetes, crossreactivity is minimal and its clinical significance is not clear. These data confirm that for reliable diagnosis of fungal allergy, representatives of both major groups must be used.

Occupational asthma caused by roasted coffee: Immunologic evidence that roasted coffee contains the same antigens as green coffee, but at a lower concentration

controlled studies, in which the numbers of patients with a positive history ranged from 24 to 37. Although reactions occurred to both non-[3-1actam antimicrobials and nonantimicrobial drugs, the frequency of these reactions was not greater in group A or B, compared with group C, which suggests that the existence of a history of penicillin allergy causing a change in prescribing habits was not a factor and that there was no significant difference in reporting patterns. Only one patient in the vasomotor rhinitis group gave a history of penicillin allergy, and this patient had no history of a reaction to non-[3-1actam drugs. Because patients with a history of penicillin allergy in this study have an incidence of reactions to non[3-1actam drugs that is equal to that found in the vasomotor rhinitis group, they do have multiple drug reactions, because they reacted to both [3-1actarn and non-[~-lactam drugs. But the risk of reacting to non-[3-1actams is not increased in either patients with a positive history of penicillin allergy or those with skin test-proven penicillin allergy, when compared with the control rhinitis group. So although this survey is also subject to the deficiencies of retrospective evaluation and limited by the preponderance of female subjects and sample size, it does suggest, in contrast to previous studies, that the development of penicillin allergy and non-[3-1actam allergy are not associated, but occur independently.

Fusarium solani: Evidence for common antigenic / allergenic determinants with other Fungi Imperfecti

Aqueous extracts of Fusarium solani and other members of the Fungi Imperfecti were evaluated for the presence of common antigenic/allergenic determinants using skin‐prick testing, radio‐allergo‐sorbent test (RAST) inhibition, and immunoelectrophoretic methods. Prevalence of skin reactivity in forty‐four atopic individuals, tested with commercially available fungal extracts, ranged from 27.3% for Alternaria tenuis to 6.8% for Penicillium notatum. No specific patterns of reactivity emerged from statistical analyses of skin test data. In contrast, RAST inhibition demonstrated common allergenic determinants. P. notatum and Aspergillus glaucus inhibited F. solani RAST by 79% and 84%, respectively. This was supported by crossed line immunoelectrophoresis; both P. notatum and A. glaucus had antigenic determinants in common with F. solani. Collectively, these studies suggest that F. solani, P. notatum, and A. glaucus have several common antigenic/allergenic determinants.

Stability Studies of Calvatia cyathiformis Basidiospore Allergens

Calvatia cyathiformis allergens in unfractionated extract (crude), and in extract sequentially fractionated by gel filtration (GF) and hydrophobic interaction chromatography (HIC) were tested for stability. C. cyathiformis allergen sources (crude, GF, HIC) were sampled immediately (0 h), or incubated at 4, 24 or 37 degrees C and then sampled at 8, 24 or 96 h. Polyacrylamide gel-isoelectric focusing immunoprints revealed 3 allergen(s) groups, or bands (Bds) with respective pI of 3.6-4.6, 6.6 and 9.3. Only Bds 3.6-4.6 were stable at 37 degrees C. At 24 degrees C, Bds 3.6-4.6 persisted to 96 h, Bd 6.6 persisted 24 h, and Bd 9.3 waned in 8 h. At 4 degrees C all 3 allergens in HIC were stable for 8 and 24 h; Bd 9.3 was reduced at 96 h. All allergen activity was labile to low pH conditions except for Bds 3.6-4.6. Proteinase K degraded Bd 9.3 more rapidly than Bd 6.6. Immunoprint patterns corresponded to the stained gels and were consistent among different sources. Bd 9.3 is very labile, but reactive with 63% of sera tested. Since 10-15% of C. cyathiformis reactors bind IgE only to Bd 9.3, this is notable variability, and significant for diagnosis and treatment.

Analysis of tobacco leaf allergens by crossed radioimmunoelectrophoresis

The reactivity of eleven ‘tobacco smoke sensitive’ and eight ‘non‐sensitive’ individuals to tobacco leaf allergens was tested by Crossed Radioimmunoelectrophoresis (CRIE). All nineteen study subjects had IgE antibodies to tobacco leaf antigens as measured by Radioallergosorbent Test (RAST) and seventeen of the nineteen individuals were atopic. Of the thirty‐seven tobacco leaf precipitins detected by Cross Immunoelectro‐phoresis (CIE), three were identified as allergens by CRIE. All nineteen subjects reacted to at least one of the three allergens detected. However, neither the intensity nor the incidence of reactivity with any of the three allergens correlated with smoking or ‘smoke sensitivity’.