title: Removal of sialic acid from mucin-like surface molecules of Trypanosoma cruzi metacyclic trypomastigotes enhances parasite-host cell interaction.
abstract: The 35/50 kDa mucin-like surface glycoprotein (gp35/50) of Trypanosoma cruzi metacyclic trypomastigotes has been implicated in mammalian cell invasion. In this study we investigated whether the sialyl residues of gp35/50 are required for interaction of parasites with target cells. After treatment with bacterial neuraminidase, the metacyclic forms (G strain) remained reactive with the monoclonal antibody (mAb) 10D8 but lost their reactivity with mAb 3C9, that recognizes sialic acid-containing epitopes on gp35/50, and entered HeLa cells in significantly higher numbers as compared to untreated controls. Resialylation of gp35/50, by incubation of parasites with T. cruzi trans-sialidase and sialyl lactose, restored the reactivity with mAb 3C9 as well as the affinity for sialic acid specific lectin. Accordingly, the rate of invasion of resialylated parasites was reduced to levels similar to those observed before desialylation. Purified G strain gp35/50, desialylated by neuraminidase treatment, bound to HeLa cells more than its sialylated counterpart. The Ca2+ signaling activity, which has been associated with cell invasion, was also determined by measuring the cytosolic Ca2+ concentration ([Ca2+]i), in HeLa cells upon interaction with sonicated extracts from untreated or neuraminidase-treated parasites, or with purified gp35/50 in its sialylated or desialylated form. Consistent with the results of cell invasion assay, the desialylated parasite preparations, as well as the sialic acid free gp35/50, induced an average elevation in [Ca2+]i significantly higher than that triggered by untreated controls. None of these effects, namely the increase in infectivity and Ca2+ signaling activity, was observed with neuraminidase-treated CL strain metacyclic trypomastigotes, which express a variant form of sialic acid gp35/50 molecule that is not recognized by mAb 10D8 and apparently is not involved in target cell invasion.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, Brazil. nyoshida.dmip@epm.br
authors.forename: N
authors.initials: N
authors.lastname: Yoshida
authors.forename: M L
authors.initials: ML
authors.lastname: Dorta
authors.forename: A T
authors.initials: AT
authors.lastname: Ferreira
authors.forename: M E
authors.initials: ME
authors.lastname: Oshiro
authors.forename: R A
authors.initials: RA
authors.lastname: Mortara
authors.forename: A
authors.initials: A
authors.lastname: Acosta-Serrano
authors.forename: S
authors.initials: S
authors.lastname: Favoreto Júnior
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D006367:HeLa Cells / Q000469:parasitology; D006801:Humans; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D009077:Mucins / Q000302:isolation & purification*; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase / Q000494:pharmacology; D015800:Protozoan Proteins / Q000302:isolation & purification*; D014349:Trypanosoma cruzi / Q000737:chemistry* / Q000187:drug effects / Q000276:immunology
chemical_list: D008562:Membrane Glycoproteins; D009077:Mucins; D015800:Protozoan Proteins; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid; D002118:Calcium