Immunoblot analysis of IgE and IgG binding antigens in extracts of mosquitos Aedes vexans, Culex tarsalis and Culiseta inornata

Abstract

Extracts of one globally distributed mosquito species (Aedes vexans) and two North American species (Culex tarsalis and Culiseta inornata) were prepared from heads and thoraxes. Proteins of the three extracts were separated by 12% SDS-PAGE and transferred to nitrocellulose membranes for immunoblotting. Immunoblotting was completed by sequential incubations of the membranes with a pooled human serum from subjects allergic to mosquito bites, monoclonal antibodies to human IgE or IgG, and goat antimouse IgG conjugate. Twelve to sixteen antigens with molecular masses ranging from 18.5 to 160 kDa were found in each extract. Nine antigens were shared by 3 species and 6 were shared by 2 species. Only 3 were species-unique. Most antigens bound to both IgE and IgG antibodies. IgE and IgG antibodies against A. vexans were studied by immunoblotting using individual serum from subjects with or without skin reactions 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. The patterns and magnitudes of IgE and IgG antibodies to the antigens varied among individuals. Very faint IgE antibodies to these antigens were found in the 2 subjects with no skin reactions, suggesting that IgE plays a role in the development of mosquito allergy.