Polyclonal human IgE antibodies of patients sensitized to one species of the Diptera family Chironomidae as well as polyclonal rabbit anti-Chi t I hyperimmune serum recognize antigenic sites present in nearly all of the 33 species of this insect family. Evolutionary distantly related genera usually show weaker antibody binding. According to the pattern of reactivity of rabbit anti-Chi t I-component III, expression of the epitopes of this molecule varies considerably in the genera Chironomus and Glyptotendipes; it appears to be almost totally absent in all other species. Of five monoclonal antibodies raised against Chi t I-component III, two recognize an epitope which is expressed in nearly all closely related species. Three monoclonal antibodies recognize epitopes which are expressed in only a few species of the same genus. Our results demonstrate the presence of common as well as species-specific epitopes in chironomid hemoglobins which behave as potent inhalant allergens.