GALACTOFURANOSE-CONTAINING GLYCOCONJUGATES OF EPIMASTIGOTE AND TRYPOMASTIGOTE FORMS OF TRYPANOSOMA-CRUZI

Antiserum to LPPG, a lipopeptidophosphoglycan originally described on the surface of Trypanosoma cruzi epimastigotes of the Y strain, and antibodies to furanoic galactose (gal(f)) were obtained in rabbits. A micromethod for the extraction and purification of LPPG from a limited amount of parasites is described. Analysis by Western blots of the purified glycoconjugate probed with both antisera confirmed the presence of gal(f)-containing LPPG-like molecules in 10 different strains and clones of T. cruzi. An analogous approach indicated that trypomastigotes also contain LPPG-like components. Quantitation experiments allowed to calculate an average value of 1.0 X 10(7) LPPG molecules per epimastigote cell and 0. 16 X 10(7) LPPG-like molecules per trypomastigote cell. Immunoelectron microscopy has shown a homogeneous distribution of LPPG on the surface of epimastigotes. The trypomastigote population, however, is highly heterogeneous with no more than 15% of the parasites being labeled by the anti-LPPG serum. Intense labeling has also been found in vesicles inside the epimastigote and trypomastigote forms. The distribution of gal(f) epitopes among glycoconjugates of epimastigotes and trypomastigotes was further investigated. It was shown that gal(f) units in epimastigotes are bound to low molecular mass compounds which co-migrate with LPPG whereas in trypomastigotes they have been found in both low molecular mass LPPG-like molecules and glycoproteins of 80-90 kDa. Direct chemical evidence for the presence of gal(f) residues in the N-linked oligosaccharide chains of these surface glycoproteins has been obtained. Finally, the natural antigenicity of LPPG and gal(f) in chronic Chagas' disease was investigated. It was found that all chronic chagasic sera investigated recognize this glycoconjugate and that an important part of such recognition can be attributed to gal(f) residues. Furthermore, no correlation among reactivity to LPPG, strain zymodeme and clinical forms of the disease was found.