Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1998 Feb 15;330(Pt 1):505–511. doi: 10.1042/bj3300505

Infectivity of Trypanosoma cruzi strains is associated with differential expression of surface glycoproteins with differential Ca2+ signalling activity.

R C Ruiz 1, S Favoreto Jr 1, M L Dorta 1, M E Oshiro 1, A T Ferreira 1, P M Manque 1, N Yoshida 1
PMCID: PMC1219166  PMID: 9461549

Abstract

Mammalian cell invasion assays, using metacyclic trypomastigotes of Trypanosoma cruzi G and CL strains, showed that the CL strain enters target cells in several-fold higher numbers as compared with the G strain. Analysis of expression of surface glycoproteins in metacyclic forms of the two strains by iodination, immunoprecipitation and FACS, revealed that gp90, undetectable in the CL strain, is one of the major surface molecules in the G strain, that expression of gp82 is comparable in both strains and that gp35/50 is expressed at lower levels in the CL strain. Purified gp90 and gp35/50 bound more efficiently than gp82 to cultured HeLa cells. However, the intensity of the Ca2+ response triggered in HeLa cells by gp82 was significantly higher than that induced by gp35/50 or gp90. Most of the Ca2+ signalling activity of the metacyclic extract towards HeLa cells was due to gp82 and was inhibitable by gp82-specific monoclonal antibody 3F6. Ca2+ mobilization was also triggered in metacyclic trypomastigotes by host-cell components; it was mainly gp82-mediated and more intense in the CL than in the G strain. We propose that expression of gp90 and gp35/50 at high levels impairs binding of metacyclic forms to host cells through productive gp82-mediated interaction, which leads to the target-cell and parasite Ca2+ mobilization required for invasion. Analysis of metacyclic forms of eight additional T. cruzi strains corroborated the inverse correlation between infectivity and expression of gp90 and gp35/50.

Full Text

The Full Text of this article is available as a PDF (419.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Araguth M. F., Rodrigues M. M., Yoshida N. Trypanosoma cruzi metacyclic trypomastigotes: neutralization by the stage-specific monoclonal antibody 1G7 and immunogenicity of 90 kD surface antigen. Parasite Immunol. 1988 Nov;10(6):707–712. doi: 10.1111/j.1365-3024.1988.tb00256.x. [DOI] [PubMed] [Google Scholar]
  2. Araya J. E., Cano M. I., Yoshida N., da Silveira J. F. Cloning and characterization of a gene for the stage-specific 82-kDa surface antigen of metacyclic trypomastigotes of Trypanosoma cruzi. Mol Biochem Parasitol. 1994 May;65(1):161–169. doi: 10.1016/0166-6851(94)90124-4. [DOI] [PubMed] [Google Scholar]
  3. BRENER Z., CHIARI E. VARIA C OES MORFOL'OGICAS OBSERVADAS EM DIFERENTES AMOSTRAS DE TRYPANOSOMA CRUZI. Rev Inst Med Trop Sao Paulo. 1963 Sep-Oct;5:220–224. [PubMed] [Google Scholar]
  4. Barbiéri C. L., Giorgio S., Merjan A. J., Figueiredo E. N. Glycosphingolipid antigens of Leishmania (Leishmania) amazonensis amastigotes identified by use of a monoclonal antibody. Infect Immun. 1993 May;61(5):2131–2137. doi: 10.1128/iai.61.5.2131-2137.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barr S. C., Han W., Andrews N. W., Lopez J. W., Ball B. A., Pannabecker T. L., Gilmour R. F., Jr A factor from Trypanosoma cruzi induces repetitive cytosolic free Ca2+ transients in isolated primary canine cardiac myocytes. Infect Immun. 1996 May;64(5):1770–1777. doi: 10.1128/iai.64.5.1770-1777.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burleigh B. A., Andrews N. W. A 120-kDa alkaline peptidase from Trypanosoma cruzi is involved in the generation of a novel Ca(2+)-signaling factor for mammalian cells. J Biol Chem. 1995 Mar 10;270(10):5172–5180. doi: 10.1074/jbc.270.10.5172. [DOI] [PubMed] [Google Scholar]
  7. CAMARGO E. P. GROWTH AND DIFFERENTIATION IN TRYPANOSOMA CRUZI. I. ORIGIN OF METACYCLIC TRYPANOSOMES IN LIQUID MEDIA. Rev Inst Med Trop Sao Paulo. 1964 May-Jun;6:93–100. [PubMed] [Google Scholar]
  8. Deane M. P., Kloetzel J. Lack of protection against Trypanosoma cruzi by multiple doses of T. lewisi culture forms. A discussion on some strains of "lewisi". Exp Parasitol. 1974 Jun;35(3):406–410. doi: 10.1016/0014-4894(74)90046-0. [DOI] [PubMed] [Google Scholar]
  9. Dorta M. L., Ferreira A. T., Oshiro M. E., Yoshida N. Ca2+ signal induced by Trypanosoma cruzi metacyclic trypomastigote surface molecules implicated in mammalian cell invasion. Mol Biochem Parasitol. 1995 Jul;73(1-2):285–289. doi: 10.1016/0166-6851(94)00123-5. [DOI] [PubMed] [Google Scholar]
  10. Franco F. R., Paranhos-Bacallà G. S., Yamauchi L. M., Yoshida N., da Silveira J. F. Characterization of a cDNA clone encoding the carboxy-terminal domain of a 90-kilodalton surface antigen of Trypanosoma cruzi metacyclic trypomastigotes. Infect Immun. 1993 Oct;61(10):4196–4201. doi: 10.1128/iai.61.10.4196-4201.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  12. Güther M. L., de Almeida M. L., Yoshida N., Ferguson M. A. Structural studies on the glycosylphosphatidylinositol membrane anchor of Trypanosoma cruzi 1G7-antigen. The structure of the glycan core. J Biol Chem. 1992 Apr 5;267(10):6820–6828. [PubMed] [Google Scholar]
  13. Heise N., de Almeida M. L., Ferguson M. A. Characterization of the lipid moiety of the glycosylphosphatidylinositol anchor of Trypanosoma cruzi 1G7-antigen. Mol Biochem Parasitol. 1995 Mar;70(1-2):71–84. doi: 10.1016/0166-6851(95)00009-p. [DOI] [PubMed] [Google Scholar]
  14. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  15. Mello D. A., Borges M. M., Chiarini L. H. Crescimento e diferenciaço "in vitro" de cepas de Trypanosoma cruzi, isolada de animais silvestres. Rev Saude Publica. 1980 Dec;14(4):569–581. doi: 10.1590/s0034-89101980000400012. [DOI] [PubMed] [Google Scholar]
  16. Mello D. A., Borges M. M. Primeiro encontro do Triatoma costalimai naturalmente infectado pelo Trypanosoma cruzi: estudo de aspectos biológicos da amostra isolada. Mem Inst Oswaldo Cruz. 1981 Jan-Mar;76(1):61–69. doi: 10.1590/s0074-02761981000100007. [DOI] [PubMed] [Google Scholar]
  17. Moreno S. N., Silva J., Vercesi A. E., Docampo R. Cytosolic-free calcium elevation in Trypanosoma cruzi is required for cell invasion. J Exp Med. 1994 Oct 1;180(4):1535–1540. doi: 10.1084/jem.180.4.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mortara R. A., Araguth M. F., Yoshida N. Reactivity of stage-specific monoclonal antibody 1G7 with metacyclic trypomastigotes of Trypanosoma cruzi strains: lytic property and 90,000 mol. wt surface antigen polymorphism. Parasite Immunol. 1988 Jul;10(4):369–378. doi: 10.1111/j.1365-3024.1988.tb00227.x. [DOI] [PubMed] [Google Scholar]
  19. Mortara R. A., da Silva S., Araguth M. F., Blanco S. A., Yoshida N. Polymorphism of the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi metacyclic trypomastigotes. Infect Immun. 1992 Nov;60(11):4673–4678. doi: 10.1128/iai.60.11.4673-4678.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. PIZZI T., RUBIO M., PRAGER R., SILVA R. Acción de la cortisona en la infección experimental por Trypanosoma cruzi; communicación preliminar. Bol Inf Parasit Chil. 1952 Apr-Jun;7(2):22–24. [PubMed] [Google Scholar]
  21. Ramirez M. I., Ruiz R. de C., Araya J. E., Da Silveira J. F., Yoshida N. Involvement of the stage-specific 82-kilodalton adhesion molecule of Trypanosoma cruzi metacyclic trypomastigotes in host cell invasion. Infect Immun. 1993 Sep;61(9):3636–3641. doi: 10.1128/iai.61.9.3636-3641.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ruiz R. de C., Rigoni V. L., Gonzalez J., Yoshida N. The 35/50 kDa surface antigen of Trypanosoma cruzi metacyclic trypomastigotes, an adhesion molecule involved in host cell invasion. Parasite Immunol. 1993 Feb;15(2):121–125. doi: 10.1111/j.1365-3024.1993.tb00591.x. [DOI] [PubMed] [Google Scholar]
  23. Santori F. R., Dorta M. L., Juliano L., Juliano M. A., da Silveira J. F., Ruiz R. C., Yoshida N. Identification of a domain of Trypanosoma cruzi metacyclic trypomastigote surface molecule gp82 required for attachment and invasion of mammalian cells. Mol Biochem Parasitol. 1996 Jun;78(1-2):209–216. doi: 10.1016/s0166-6851(96)02626-6. [DOI] [PubMed] [Google Scholar]
  24. Schenkman S., Ferguson M. A., Heise N., de Almeida M. L., Mortara R. A., Yoshida N. Mucin-like glycoproteins linked to the membrane by glycosylphosphatidylinositol anchor are the major acceptors of sialic acid in a reaction catalyzed by trans-sialidase in metacyclic forms of Trypanosoma cruzi. Mol Biochem Parasitol. 1993 Jun;59(2):293–303. doi: 10.1016/0166-6851(93)90227-o. [DOI] [PubMed] [Google Scholar]
  25. Serrano A. A., Schenkman S., Yoshida N., Mehlert A., Richardson J. M., Ferguson M. A. The lipid structure of the glycosylphosphatidylinositol-anchored mucin-like sialic acid acceptors of Trypanosoma cruzi changes during parasite differentiation from epimastigotes to infective metacyclic trypomastigote forms. J Biol Chem. 1995 Nov 10;270(45):27244–27253. doi: 10.1074/jbc.270.45.27244. [DOI] [PubMed] [Google Scholar]
  26. Shenoy-Scaria A. M., Kwong J., Fujita T., Olszowy M. W., Shaw A. S., Lublin D. M. Signal transduction through decay-accelerating factor. Interaction of glycosyl-phosphatidylinositol anchor and protein tyrosine kinases p56lck and p59fyn 1. J Immunol. 1992 Dec 1;149(11):3535–3541. [PubMed] [Google Scholar]
  27. Stefanová I., Horejsí V., Ansotegui I. J., Knapp W., Stockinger H. GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254(5034):1016–1019. doi: 10.1126/science.1719635. [DOI] [PubMed] [Google Scholar]
  28. Tardieux I., Nathanson M. H., Andrews N. W. Role in host cell invasion of Trypanosoma cruzi-induced cytosolic-free Ca2+ transients. J Exp Med. 1994 Mar 1;179(3):1017–1022. doi: 10.1084/jem.179.3.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Teixeira M. M., Yoshida N. Stage-specific surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi identified by monoclonal antibodies. Mol Biochem Parasitol. 1986 Mar;18(3):271–282. doi: 10.1016/0166-6851(86)90085-x. [DOI] [PubMed] [Google Scholar]
  30. Thomas P. M., Samelson L. E. The glycophosphatidylinositol-anchored Thy-1 molecule interacts with the p60fyn protein tyrosine kinase in T cells. J Biol Chem. 1992 Jun 15;267(17):12317–12322. [PubMed] [Google Scholar]
  31. Wilkowsky S. E., Wainszelbaum M. J., Isola E. L. Trypanosoma cruzi: participation of intracellular Ca2+ during metacyclic trypomastigote-macrophage interaction. Biochem Biophys Res Commun. 1996 May 15;222(2):386–389. doi: 10.1006/bbrc.1996.0753. [DOI] [PubMed] [Google Scholar]
  32. Yakubu M. A., Majumder S., Kierszenbaum F. Changes in Trypanosoma cruzi infectivity by treatments that affect calcium ion levels. Mol Biochem Parasitol. 1994 Jul;66(1):119–125. doi: 10.1016/0166-6851(94)90042-6. [DOI] [PubMed] [Google Scholar]
  33. Yoshida N., Araya J. E., da Silveira J. F., Giorgio S. Trypanosoma cruzi: antibody production and T cell response induced by stage-specific surface glycoproteins purified from metacyclic trypomastigotes. Exp Parasitol. 1993 Dec;77(4):405–413. doi: 10.1006/expr.1993.1100. [DOI] [PubMed] [Google Scholar]
  34. Yoshida N., Blanco S. A., Araguth M. F., Russo M., González J. The stage-specific 90-kilodalton surface antigen of metacyclic trypomastigotes of Trypanosoma cruzi. Mol Biochem Parasitol. 1990 Feb;39(1):39–46. doi: 10.1016/0166-6851(90)90006-8. [DOI] [PubMed] [Google Scholar]
  35. Yoshida N., Mortara R. A., Araguth M. F., Gonzalez J. C., Russo M. Metacyclic neutralizing effect of monoclonal antibody 10D8 directed to the 35- and 50-kilodalton surface glycoconjugates of Trypanosoma cruzi. Infect Immun. 1989 Jun;57(6):1663–1667. doi: 10.1128/iai.57.6.1663-1667.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yoshida N. Surface antigens of metacyclic trypomastigotes of Trypanosoma cruzi. Infect Immun. 1983 May;40(2):836–839. doi: 10.1128/iai.40.2.836-839.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. de Almeida M. L., Heise N. Proteins anchored via glycosylphosphatidylinositol and solubilizing phospholipases in Trypanosoma cruzi. Biol Res. 1993;26(1-2):285–312. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES