Purification and characterization of an allergenic monomeric hemoglobin from a chironomid distributed worldwide, Polypedium nubifer.
The Pol n component MV, a potent experimental allergen for mice, was purified to homogeneity from extracts of a chironomid distributed worldwide, Polypedium nubifer (PN). The Pol n I component MV was shown to have cross-reactivity to hemoglobins (Hb) derived from all species of chironomids tested. Determination of the amino acid sequence of the first 37 N-terminal residues revealed that it had 30-59% homology to Hb of an European chironomid, Chironomus thummi thummi, which had been known as an important allergen for humans. By Western blot analysis, we showed that sera from asthmatic patients, which had positively reacted to the extract of the adult PN midge, bound to the purified Pol n I component MV. Furthermore, using rabbit polyclonal antibodies raised against synthetic polypeptides corresponding to the N-terminal residues, it was demonstrated that the N-terminal amino acid sequence between position 15 and 35 contained antigenic epitope(s) for human IgE. The results indicate that the Pol n I component MV is an allergen for human beings as well as for mice, and useful as a diagnostic tool for chironomid allergy.
Kawai
K
K
Department of Immunology, Toyama Medical and Pharmaceutical University, Japan.
Tagoh
H
H
Yoshizaki
K
K
Murakami
G
G
Muraguchi
A
A
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D001249:Asthma / Q000276:immunology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002648:Child; D002683:Chironomidae / Q000276:immunology*; D005260:Female; D006454:Hemoglobins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry; D007814:Larva / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar
D000485:Allergens; D006454:Hemoglobins; C100459:Pol n I component MV; D007073:Immunoglobulin E