title: Mucin-like molecules form a negatively charged coat that protects Trypanosoma cruzi trypomastigotes from killing by human anti-alpha-galactosyl antibodies.
abstract: In the presence of sialic acid donors Trypanosoma cruzi acquires up to 10(7) sialic acid residues on its surface, in a reaction catalyzed by its unique trans-sialidase. Most of these sialic acid residues are incorporated into mucin-like glycoproteins. To further understand the biological role of parasite sialylation, we have measured the amount of mucin in this parasite. We found that both epimastigote and trypomastigote forms have the same number of mucin molecules per surface area, although trypomastigotes have less than 10% of the amount of glycoinositol phospholipids, the other major surface glycoconjugate of T. cruzi. Based on the estimated surface area of each mucin, we calculated that these molecules form a coat covering the entire trypomastigote cell. The presence of the surface coat is shown by transmission electron microscopy of Ruthenium Red-stained parasites. The coat was revealed by binding of antibodies isolated from Chagasic patients that react with high affinity to alpha-galactosyl epitopes present in the mucin molecule. When added to the trypomastigote, these antibodies cause an extensive structural perturbation of the parasite coat with formation of large blebs, ultimately leading to parasite lysis. Interestingly, lysis is decreased if the mucin coat is heavily sialylated. Furthermore, addition of MgCl2 reverses the protective effect of sialylation, suggesting that the sialic acid negative charges stabilize the surface coat. Inhibition of sialylation by anti-trans-sialidase antibodies, found in immunized animals, or human Chagasic sera, also increase killing by anti-alpha-galactosyl antibodies. Therefore, the large amounts of sialylated mucins, forming a surface coat on infective trypomastigote forms, have an important structural and protective role.
authors.affiliation: Department of Microbiologia, Imunologia e Parasitologia, UNIFESP, R. Botucatu 862 8A, São Paulo, SP, Brazil.
authors.forename: V L
authors.initials: VL
authors.lastname: Pereira-Chioccola
authors.forename: A
authors.initials: A
authors.lastname: Acosta-Serrano
authors.forename: I
authors.initials: I
authors.lastname: Correia de Almeida
authors.forename: M A
authors.initials: MA
authors.lastname: Ferguson
authors.forename: T
authors.initials: T
authors.lastname: Souto-Padron
authors.forename: M M
authors.initials: MM
authors.lastname: Rodrigues
authors.forename: L R
authors.initials: LR
authors.lastname: Travassos
authors.forename: S
authors.initials: S
authors.lastname: Schenkman
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000633:toxicity*; D000953:Antigens, Protozoan / Q000276:immunology*; D014355:Chagas Disease / Q000276:immunology / Q000469:parasitology; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate; D009077:Mucins / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D013499:Surface Properties; D014312:Trisaccharides / Q000276:immunology*; D014349:Trypanosoma cruzi / Q000254:growth & development* / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017261:Glycosylphosphatidylinositols; D009077:Mucins; D014312:Trisaccharides; C076569:alpha-galactosyl epitope; D019158:N-Acetylneuraminic Acid