IMMUNOBLOT ANALYSIS OF IGE AND IGG BINDING ANTIGENS IN EXTRACTS OF MOSQUITOS AEDES-VEXANS, CULEX-TARSALIS AND CULISETA-INORNATA

Extracts of one globally distributed mosquito species (Aedes vexans) a nd two North American species (Culex tarsalis and Culiseta inornata) w ere prepared from heads and thoraxes. Proteins of the three extracts w ere separated by 12% SDS-PAGE and transferred to nitrocellulose membra nes for immunoblotting. Immunoblotting was completed by sequential inc ubations of the membranes with a pooled human serum from subjects alle rgic to mosquito bites, monoclonal antibodies to human IgE or IgG, and goat antimouse IgG conjugate. Twelve to sixteen antigens with molecul ar masses ranging from 18.5 to 160 kDa were found in each extract. Nin e antigens were shared by 3 species and 6 were shared by 2 species. On ly 3 were species-unique. Most antigens bound to both IgE and IgG anti bodies. IgE and IgG antibodies against A. vexans were studied by immun oblotting using individual serum from subjects with or without skin re actions to A. vexans bites. All 3 subjects with severe skin reactions had strong IgE and IgG antibodies to 32.5, 40, and 50 kDa proteins. Th e patterns and magnitudes of IgE and IgG antibodies to the antigens va ried among individuals. Very faint IgE antibodies to these antigens we re found in the 2 subjects with no skin reactions, suggesting that IgE plays a role in the development of mosquito allergy.