Monoclonal antibodies against the flagellar fraction of epimastigotes of Trypanosoma cruzi: immunoprotection against metacyclic trypomastigotes obtained by immunization of mice with an affinity-purified antigen

Abstract

Subcellular fractions of Trypanosoma cruzi (epimastigotes) were assayed in their capacity to induce protective or aggressive effects in experimental animals. The flagellar fraction showed the best immunoprotective properties without tissular aggression. Monoclonal antibodies were prepared from mice immunized with this fraction. One of them, FCH-F8-4, was able to neutralize the infectivity of bloodstream trypomastigotes, to produce complement-mediated lysis on cell culture-derived trypomastigotes and to recognize the surface of trypomastigotes and epimastigotes by immunofluorescence. FCH-F8-4 reacted in Western blotting with several epimastigote proteins ranging from 50 to 150 kDa, showing a more intense reactivity with 4 bands while it reacted with two molecules on trypomastigote preparations (15 and 48 kDa). Purified antibody was coupled to CNBr-activated Sepharose and used to purify antigens from epimastigote extracts. These antigens were used to immunize BALB/c mice in the presence of Bordetella pertussis as adjuvant. Animals were protected against a challenge with 10 3 metacyclic forms of T. cruzi (Tulahuén strain). Only 40% of immunized mice presented detectable parasites in blood after challenge. Parasitemia decreased 90% in relation to controls in those animals. Survival of immunized mice was 100% in all immunoprotection experiments. These results suggest that the epitope recognized by FCH-F8-4 present in the purified antigens keeps the protective characteristics of flagellar fraction and could be a candidate for the development of a vaccine against T. cruzi infection.