Role of sialic acid in the resistance of Trypanosoma cruzi trypomastigotes to complement.
Trypomastigotes of Trypanosoma cruzi, mammalian infective forms of the parasite, express an unusual cell surface trans-sialidase. This enzyme enables the parasite to rapidly sialylate its surface when supplied with alpha(2,3)-linked sialic acid from glycoconjugates in serum or on cell surfaces. Here we used a novel fluorescence-based, trypomastigote lysis assay to evaluate the role of sialic acid on the parasite's plasma membrane in providing protection against the complement cascade. Trypomastigotes were desialylated, and sialic acid removal was confirmed by a chemical assay and also by flow cytometry with the use of a mAb that recognizes a T. cruzi-sialylated epitope. Compared with sialylated trypomastigotes, which were completely refractory to lysis by human serum, only about 5% of the desialylated trypomastigotes were lysed by complement. However, further analysis revealed that the desialylated parasites had been resialylated during exposure to serum complement. Next we incubated desialylated trypomastigotes with samples of desialylated human serum. Although the sialidase-treated serum retained its full hemolytic activity, lysis of trypomastigotes increased only from 5 to 24%. This increase correlated with an enhanced deposition of complement protein C3 on the parasite surface. The ratio of C3b to lytically inactive iC3b was increased for desialylated, compared with sialylated, parasites. We conclude that although parasite sialic acid promotes C3b cleavage into iC3b, this mechanism alone does not account for the robust resistance of these parasites to complement lysis.
Tomlinson
S
S
Michael Heidelberger Division of Immunology, Kaplan Cancer Center, Department of Pathology, New York University Medical Center, NY 10016.
Pontes de Carvalho
L C
LC
Vandekerckhove
F
F
Nussenzweig
V
V
D000818:Animals; D003176:Complement C3 / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D009439:Neuraminidase / Q000378:metabolism; D012794:Sialic Acids / Q000276:immunology*; D014349:Trypanosoma cruzi / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology
D003176:Complement C3; D012794:Sialic Acids; D009439:Neuraminidase; D014357:Trypsin