Schistosoma mansoni: identification and characterization of schistosomula polypeptides.
Schistosomula proteins separated by a two-dimensional (NEPHGE) gel system identify 94 major silver-stained polypeptides. When compared to polypeptides similarly separated from cercariae and adult worms; cercariae share the same polypeptides as schistosomula, adult worms share ca. 60% of the polypeptides. A group of five schistosomula polypeptides 15-31 kDa (apparent pI 8.2-8.9) was not found in adult worm extracts. To identify which polypeptides were immunogens, Western blots of the NEPHGE gels were probed with sera either from humans with chronic schistosomiasis or from mice vaccinated with irradiated cercariae. For characterization studies, polyclonal antibodies were made against the five schistosomula-specific and selected immunogenic polypeptides by immunizing mice with silver-stained spots removed from NEPHGE gels. We show that the polyclonal serum against a polypeptide of 12.5 kDa and an apparent pI of 6.70 mediated complement and eosinophil-dependent killing of schistosomula in an in vitro assay. Epitopes recognized by antibody against the 12.5-kDa polypeptide show a diffuse distribution and are found on flame cells of the excretory system of the schistosomula.
Miller
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Department of Microbiology, State University of New York, Buffalo 14214.
Rekosh
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LoVerde
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D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000947:Antigens, Helminth / Q000302:isolation & purification*; D015153:Blotting, Western; D003602:Cytotoxicity, Immunologic; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010455:Peptides / Q000302:isolation & purification*; D012550:Schistosoma mansoni / Q000032:analysis* / Q000276:immunology; D012555:Schistosomiasis mansoni / Q000097:blood*
D000906:Antibodies; D000947:Antigens, Helminth; D010455:Peptides