Involvement of the stage-specific 82-kilodalton adhesion molecule of Trypanosoma cruzi metacyclic trypomastigotes in host cell invasion.

This study provides several pieces of evidence indicating that 3F6-Ag, identified by monoclonal antibody (MAb) 3F6 as a stage-specific glycoprotein of approximately 82 kDa on the surface of metacyclic trypomastigotes of different Trypanosoma cruzi strains, promotes the entry of parasites into host cells through a ligand-receptor type interaction. First, invasion of Vero cells by metacyclic trypomastigotes of both CL and Tulahuen strains was significantly inhibited by MAb 3F6 or its Fab fragments. Second, purified 3F6-Ag bound to Vero cells in a dose-dependent and saturable fashion. Third, soluble 3F6-Ag reduced the infection of Vero cells by metacyclic forms of CL and Tulahuen strains by 90 to 97 and 50%, respectively. Unrelated proteins, as well as extracellular matrix components, such as heparan sulfate and collagen, had no effect. Our studies also show that in the Tulahuen strain, 10D8-Ag, a 35/50-kDa glycoprotein identified by MAb 10D8, participates in target cell invasion, confirming previous observations, but the variant form of 10D8-Ag expressed by highly invasive CL strain metacyclic trypomastigotes appears to be irrelevant. Overall, our results indicate that the surface components of T. cruzi metacyclic trypomastigotes involved in the process of host cell penetration are developmentally regulated molecules, such as 3F6-Ag and 10D8-Ag, that have no counterpart in blood- or tissue culture-derived trypomastigotes.

The 35/50 kDa surface antigen of Trypanosoma cruzi metacyclic trypomastigotes, an adhesion molecule involved in host cell invasion.

We have previously shown that monoclonal antibodies directed to the 90 kDa glycoprotein and the 35/50 kDa glycoconjugate, present on the surface of Trypanosoma cruzi metacyclic trypomastigotes, inhibited host cell invasion. Here we investigated whether these molecules could be the ligands for the target cell receptor. Binding assays were performed by incubating Vero cells with sonicated parasite extract. Detection of bound parasite components was carried out by using monoclonal antibodies (MoAbs) IG7 and 10D8, which recognize the 90 kDa and the 35/50 kDa antigens respectively. These experiments revealed that the 35/50 kDa glycoconjugate of metacyclic forms, but not the 1G7-reactive antigen, binds to Vero cells. The purified 35/50 kDa antigen bound to Vero cells and inhibited the entry of metacyclic forms in a dose-dependent manner. Although to a lesser extent, an immunologically related 35/50 kDa antigen of non-infective epimastigotes also bound to Vero cells but it was unable to inhibit parasite penetration at a concentration (100 micrograms/ml) in which metacyclic antigen exhibited more than 60% inhibition. All these data suggest that the metacyclic 35/50 kDa surface glycoconjugate is a ligand to the host cell in the process of T. cruzi invasion.

Trypanosoma cruzi: course of infection in platelets-depleted mice.

The effect of platelet depletion on the course of Trypanosoma cruzi infection in BALB/c mice was investigated. Thrombocytopenia was achieved by inoculation of rabbit anti-platelet IgG during the parasitemic phase of the infection. The number of parasites in the blood of anti-platelet IgG treated was significantly higher than that of non-treated control mice, during the phase of high parasitemia. Cumulative mortality of platelet-depleted mice was consistently but not significantly higher than that of control mice up to the 32nd day of infection; from the 33rd day on they were equivalent, no mortalities occurring from then on, until observations were discontinued on the 60th day. These results suggest that platelets participate of the mechanisms of parasites removal from the bloodstream, but do not have an effective role in the mechanisms of defence against T. cruzi, during the acute phase of infection.