Immunogenicity of the recombinant SAPA protein of Trypanosoma cruzi for mice.

The humoral and cellular immune responses induced by the recombinant SAPA (shed acute phase antigen) of Trypanosoma cruzi were studied in mice and correlated with the immunologic control of parasitemia. The immunizing schedule used consisted of 2 weekly injections of 50 micrograms glutathione-S-transferase (GST)-SAPA in Freund's adjuvant. Specific alpha GST-SAPA antibodies were detected by enzyme-linked immunosorbent assay 1 wk after each antigen dose, the concentration of antibodies after the second injection being 30-fold higher than after the first. Immediate- (ITH) and delayed-type hypersensitivity (DTH) reactions were observed as footpad swelling after injecting 50 micrograms GST-SAPA in preimmunized mice as compared to naive controls. Adoptive transfer experiments indicated that these cutaneous reactions were mediated by lymphoid cells and not by serum. Both humoral and cellular responses were specific for the GST-SAPA antigen and did not cross-react with either the GST or the recombinant GST-1 T. cruzi antigen. Immunized mice that had developed high levels of antibody and DTH reaction to GST-SAPA were able to control the level of parasitemia after challenge with 10(3) blood trypomastigotes. The levels of parasitemia obtained were lowered to about 1/3 (P < 0.05) and mortality at day 60 was reduced from 67 to 25% (P = 0.085). Comparison of this immunizing method with other schedules involving more injections or higher antigen doses indicates that control of parasitemia can be obtained with low amounts of antigen and seems to be associated with the development of DTH.

Immunoassay with recombinant Trypanosoma cruzi antigens potentially useful for screening donated blood and diagnosing Chagas disease.

A new enzyme immunoassay (EIA), the Dia Kit Bio-Chagas assay (Gador S.A.), is potentially useful for detecting antibodies to Trypanosoma cruzi in the diagnosis of infected individuals and the screening of blood in blood banks. The EIA is carried out on test strips of plastic backing covered with a nitrocellulose membrane to which a mixture of recombinant T. cruzi antigens 1, 2, shed acute-phase antigen, 13, and 30 has been applied as a horizontal line. A horizontal line of human IgG is included to monitor the test procedure. The test strips exhibited homogeneity in the adsorption of the mixture of recombinant antigens (CV = 6.0%) and the human IgG (CV = 8.6%). The EIA results obtained with sera positive by xenodiagnosis showed 100% agreement between both types of tests; tested against sera with positive and negative matched results of indirect hemagglutination, immunofluorescence, and ELISA, the EIA results agreed for 99.1% (347 of 350) and 99.6% (299 of 300) of the samples, respectively.