Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies

Teixeira, Marta Maria Geraldes; Yoshida, Nobuko

doi:10.1016/0166-6851(86)90085-x

Cited by 160 publications

(151 citation statements)

References 22 publications

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“…Liver infusion tryptose medium (13) and Graces medium (Gibco, Gaithersburg, MD, USA) were used, respectively, to grow parasites and to obtain cultures enriched in metacyclic forms. Metacyclic trypomastigotes were purified by passage through a DEAE-cellulose column, as described (14). HeLa cells, the human carcinoma-derived epithelial cells, and human leukemic K562 cells (15) were grown at 37 o C in Dulbeccos minimum essential medium (DMEM) supplemented with 10% fetal calf serum (FCS), streptomycin (100 µg/ml) and penicillin (100 U/ml) in a humidified 5% CO 2 atmosphere.…”

Section: T Cruzi Mammalian Cells and Cell Invasion Assaymentioning

confidence: 99%

Signal transduction induced in Trypanosoma cruzi metacyclic trypomastigotes during the invasion of mammalian cells

Yoshida

Favoreto

Ferreira

et al. 2000

Braz J Med Biol Res

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Penetration of Trypanosoma cruzi into mammalian cells depends on the activation of the parasites protein tyrosine kinase and on the increase in cytosolic Ca 2+ concentration. We used metacyclic trypomastigotes, the T. cruzi developmental forms that initiate infection in mammalian hosts, to investigate the association of these two events and to identify the various components of the parasite signal transduction pathway involved in host cell invasion. We have found that i) both the protein tyrosine kinase activation, as measured by phosphorylation of a 175-kDa protein (p175), and Ca 2+ mobilization were induced in the metacyclic forms by the HeLa cell extract but not by the extract of T. cruzi-resistant K562 cells; ii) treatment of parasites with the tyrosine kinase inhibitor genistein blocked both p175 phosphorylation and the increase in cytosolic Ca 2+ concentration; iii) the recombinant protein J18, which contains the full-length sequence of gp82, a metacyclic stage surface glycoprotein involved in target cell invasion, interfered with tyrosine kinase and Ca 2+ responses, whereas the monoclonal antibody 3F6 directed at gp82 induced parasite p175 phosphorylation and Ca 2+ mobilization; iv) treatment of metacyclic forms with phospholipase C inhibitor U73122 blocked Ca 2+ signaling and impaired the ability of the parasites to enter HeLa cells, and v) drugs such as heparin, a competitive IP 3 -receptor blocker, caffeine, which affects Ca 2+ release from IP 3 -sensitive stores, in addition to thapsigargin, which depletes intracellular Ca 2+ compartments and lithium ion, reduced the parasite infectivity. Taken together, these data suggest that protein tyrosine kinase, phospholipase C and IP 3 are involved in the signaling cascade that is initiated on the parasite cell surface by gp82 and leads to Ca 2+ mobilization required for target cell invasion.

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Section: T Cruzi Mammalian Cells and Cell Invasion Assaymentioning

confidence: 99%

Signal transduction induced in Trypanosoma cruzi metacyclic trypomastigotes during the invasion of mammalian cells

Yoshida

Favoreto

Ferreira

et al. 2000

Braz J Med Biol Res

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“…Its expression is thus inversely correlated with the parasite's infectivity (Ruiz et al, 1998;Cortez et al, 2006). Previous in vitro studies showed that GP82 and GP90 genes are preferentially transcribed and expressed in the metacyclic trypomastigote stage (Teixeira and Yoshida, 1986;Araya et al, 1994;Carmo et al, 1999Carmo et al, , 2002. What is not known is the transcription pattern of GP82 and GP90 genes, as well as the expression of the corresponding proteins, during metacyclogenesis in triatomines infected with T. cruzi.…”

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confidence: 90%

“…Metacyclic trypomastigotes express two major stage-specific surface glycoproteins called GP82 and GP90, which have been implicated in the invasion of mammalian host cells (Teixeira and Yoshida, 1986;Yoshida, 2006). GP82 is a cell adhesion molecule that plays a key role in host cell invasion (Ramirez et al, 1993) by inducing Ca 2+ response in target cells and in the parasite (Ruiz et al, 1998), an event that is required for parasite internalization (Moreno et al, 1994;Tardieux et al, 1994;Dorta et al, 1995 (Ruiz et al, 1998) and functions as a down-regulator of the target-cell invasion (Málaga and Yoshida, 2001).…”

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confidence: 99%

Expression of GP82 and GP90 surface glycoprotein genes of Trypanosoma cruzi during in vivo metacyclogenesis in the insect vector Rhodnius prolixus

et al. 2008

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“…Serum samples from 16 patients were studied. In some experiments the ' sera were tested individually (CS 1 to CS 5) or in pools (PI: CS 6, 7, 8, 9 and P2: CS 10,11,12,13,14,15,16).…”

Section: Methodsmentioning

confidence: 99%

Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

Takehara

Cardoso

Silva

et al. 1988

Rev. Inst. Med. trop. S. Paulo

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SUMMARYSera of Chaga's disease patients containing anti-T. cruzi lytic antibodies were submitted to affinity chromatography using Sepharose 4B conjugated with antigen extracted from epimastigote or trypomastigote forms of the parasite. Epimastigotes were obtained from culture at the exponential growth phase and the trypomastigotes from blood of infected and immunosuppressed mice. Antigen of both parasite forms was obtained by sonication of the parasites followed by cemrifugation. Both antigens were then conjugated to activated Sepharose 4B. Affinity chromatography was performed by passing sera from chagasic patients through an immunoadsorbent column containing either epimastigote or trypomasugote antigens. Antibodies bound to the column were eluted with cold 0,2 M glycine buffer pH 2,8. The eluted antibodies were analysed regarding their isotype and lytic activity. The results showed that anti-T. cruzi lytic antibodies present in sera from chagasic patients are mainly located in the IgG isotype and recognize epitopes present in both trypomastigote and epimastigote forms. A brief report of this work has already been published 12 .

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Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies

Cited by 160 publications

References 22 publications

Signal transduction induced in Trypanosoma cruzi metacyclic trypomastigotes during the invasion of mammalian cells

Signal transduction induced in Trypanosoma cruzi metacyclic trypomastigotes during the invasion of mammalian cells

Expression of GP82 and GP90 surface glycoprotein genes of Trypanosoma cruzi during in vivo metacyclogenesis in the insect vector Rhodnius prolixus

Lytic antibodies elicited by Trypanosoma cruzi infection recognize epitopes present on both bloodstream trypomastigote and epimastigote forms of parasite

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