Characterization of major surface and excretory-secretory immunogens ofTrypanosoma cruzitrypomastigotes and identification of potential protective antigen

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Abstract
Summary: The surface antigens ofTrypanosoma cruzitrypomastigotes were identified by immunoprecipitation and were compared with metabolically labelled excretory—secretory products (ES) released by the parasitesin vitro. A series of major immunogenic components in the ES antigens were revealed (160 kDa, 130 kDa and 80–110 kDa). The trypomastigote surface also bears the 130 kDa band and the 80–110 kDa complex. Competition experiments demonstrated the common antigenic structure of the ES and the surface antigens. Two-dimensional analysis of ES antigens immunoprecipitated by human Chagasic serum revealed several spots in the 80–110 kDa region with a wide range of isoelectric points (PI between 5·4 and 6·7). This reflects a charge heterogeneity of these polypeptides. The trypomastigote 85 kDa polypeptide was also identified in the ES antigens by using a monoclonal antibody against this antigen. Two-dimensional analysis of the 85 kDa proteins shed from the surface of trypomastigotes and immunoprecipitated by the monoclonal antibody 155D3 showed 2 major spots: a major part of the 85 kDa polypeptide was found at pH 6·5–6·6, whereas a substantial amount of the antigen was found at pH 5·7. An additional component with molecular weight of approximately 58 kDa and isoelectric points of 6·5 and 6·6, was also visualized. Detection of the 85 kDa polypeptide circulating in serum from patients with acute and chronic Chagas' disease was achieved using an enzyme-linked immunosorbent assay. In addition, the data obtained showed that a polyclonal antibody to the 85 kDa polypeptide could be used to passively induce a partial protection of Fischer rats against acute lethal infection. Thus, the antigens recognized by polyclonal antibody appear to play a role in the development of protective immunity againstT. cruzi.