IN-VITRO SYNTHESIS OF ALTERNARIA ALLERGENS AND THEIR RECOGNITION BY MURINE MONOCLONAL AND HUMAN IGE ANTIBODIES

Background: Allergic reactions to fungal allergens, including Alternar ia, are common clinical problems. Reagents used for diagnosis and ther apy of fungal allergy are complex and variable mixtures. Standardized reagents are difficult to achieve due to batch to batch variability in these biologic products, Although several Alternaria allergens have b een isolated, their production by current methods is laborious. The in troduction of molecular biology into allergy research has led to the m olecular cloning of several allergens which may culminate in improved methods of treatment. Objective: This report describes the development of techniques that will lead to the molecular cloning of Alternaria a llergens. Methods: We isolated the poly (A)(+)-messenger RNA from Alte rnaria, produced proteins in vitro and probed them for binding to muri ne monoclonal antibodies directed against Alternaria. In addition, we examined the ability of the translated proteins to bind IgE from the s era of Alternaria-sensitive individuals. Results: We synthesized at le ast 20 proteins ranging in molecular weight from 2 to 90 kD using in v itro techniques. The translated proteins were detected by both murine monoclonal and human IgE antibodies. Conclusions: Alternaria allergens can be synthesized in vitro by molecular biology techniques. Such tec hniques will be used in the development of cDNA libraries for the prod uction of Alternaria allergens by recombinant DNA methods.