Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Feb;65(2):349–357. doi: 10.1128/iai.65.2.349-357.1997

Identification of the gene family encoding the 160-kilodalton Trypanosoma cruzi complement regulatory protein.

K A Norris 1, J E Schrimpf 1, M J Szabo 1
PMCID: PMC174602  PMID: 9009282

Abstract

Trypanosoma cruzi trypomastigotes are exquisitely resistant to the lytic effects of vertebrate complement, and this characteristic contributes to the survival of the parasites in the host bloodstream. Trypomastigotes avoid complement-mediated lysis by the production of a surface glycoprotein that inhibits the formation of the alternative and classical C3 convertase, thus preventing activation and amplification of the complement cascade at the parasite surface. We have developed a monoclonal antibody to the 160-kDa T. cruzi complement regulatory protein (CRP) and describe a one-step immunoaffinity purification procedure. The CRP was purified to homogeneity and subjected to amino-terminal peptide sequence analysis. Based on the protein sequence obtained, the CRP was identified as a member of a large family of trypomastigote-specific genes, and a complete cDNA was isolated and sequenced. The complete coding sequence was cloned in Escherichia coli, and antibodies raised against the full-length recombinant protein reacted specifically with a 160-kDa protein in trypomastigote membrane protein preparations as well as with native, purified CRP. Indirect immunofluorescence revealed that the protein is uniformly expressed at the cell surfaces of trypomastigotes.

Full Text

The Full Text of this article is available as a PDF (421.9 KB).

Selected References

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

  1. Anziano D. F., Dalmasso A. P., Lelchuk R., Vásquez C. Role of complement in immune lysis of Trypanosoma cruzi. Infect Immun. 1972 Nov;6(5):860–864. doi: 10.1128/iai.6.5.860-864.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beard C. A., Wrightsman R. A., Manning J. E. Identification of monoclonal antibodies against the trypomastigote stage of Trypanosoma cruzi by use of iminobiotinylated surface polypeptides. Mol Biochem Parasitol. 1985 Aug;16(2):199–212. doi: 10.1016/0166-6851(85)90087-8. [DOI] [PubMed] [Google Scholar]
  3. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fischer E., Ouaissi M. A., Velge P., Cornette J., Kazatchkine M. D. gp 58/68, a parasite component that contributes to the escape of the trypomastigote form of T. cruzi from damage by the human alternative complement pathway. Immunology. 1988 Oct;65(2):299–303. [PMC free article] [PubMed] [Google Scholar]
  5. Jazin E. E., Bontempi E. J., Sanchez D. O., Aslund L., Henriksson J., Frasch A. C., Pettersson U. Trypanosoma cruzi exoantigen is a member of a 160 kDa gene family. Parasitology. 1995 Jan;110(Pt 1):61–69. doi: 10.1017/s0031182000081051. [DOI] [PubMed] [Google Scholar]
  6. Kipnis T. L., David J. R., Alper C. A., Sher A., da Silva W. D. Enzymatic treatment transforms trypomastigotes of Trypanosoma cruzi into activators of alternative complement pathway and potentiates their uptake by macrophages. Proc Natl Acad Sci U S A. 1981 Jan;78(1):602–605. doi: 10.1073/pnas.78.1.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Nogueira N., Bianco C., Cohn Z. Studies on the selective lysis and purification of Trypanosoma cruzi. J Exp Med. 1975 Jul 1;142(1):224–229. doi: 10.1084/jem.142.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Norris K. A., Bradt B., Cooper N. R., So M. Characterization of a Trypanosoma cruzi C3 binding protein with functional and genetic similarities to the human complement regulatory protein, decay-accelerating factor. J Immunol. 1991 Oct 1;147(7):2240–2247. [PubMed] [Google Scholar]
  10. Norris K. A., Galvão L. M., Schrimpf J. E., Cançado J. R., Krettli A. U. Humoral immune response to the Trypanosoma cruzi complement regulatory protein as an indicator of parasitologic clearance in human Chagas' disease. Infect Immun. 1994 Sep;62(9):4072–4074. doi: 10.1128/iai.62.9.4072-4074.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Norris K. A., Harth G., So M. Purification of a Trypanosoma cruzi membrane glycoprotein which elicits lytic antibodies. Infect Immun. 1989 Aug;57(8):2372–2377. doi: 10.1128/iai.57.8.2372-2377.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Norris K. A., Schrimpf J. E. Biochemical analysis of the membrane and soluble forms of the complement regulatory protein of Trypanosoma cruzi. Infect Immun. 1994 Jan;62(1):236–243. doi: 10.1128/iai.62.1.236-243.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Parodi A. J., Pollevick G. D., Mautner M., Buschiazzo A., Sanchez D. O., Frasch A. C. Identification of the gene(s) coding for the trans-sialidase of Trypanosoma cruzi. EMBO J. 1992 May;11(5):1705–1710. doi: 10.1002/j.1460-2075.1992.tb05221.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pereira M. E., Mejia J. S., Ortega-Barria E., Matzilevich D., Prioli R. P. The Trypanosoma cruzi neuraminidase contains sequences similar to bacterial neuraminidases, YWTD repeats of the low density lipoprotein receptor, and type III modules of fibronectin. J Exp Med. 1991 Jul 1;174(1):179–191. doi: 10.1084/jem.174.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rimoldi M. T., Sher A., Heiny S., Lituchy A., Hammer C. H., Joiner K. Developmentally regulated expression by Trypanosoma cruzi of molecules that accelerate the decay of complement C3 convertases. Proc Natl Acad Sci U S A. 1988 Jan;85(1):193–197. doi: 10.1073/pnas.85.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schenkman S., Eichinger D., Pereira M. E., Nussenzweig V. Structural and functional properties of Trypanosoma trans-sialidase. Annu Rev Microbiol. 1994;48:499–523. doi: 10.1146/annurev.mi.48.100194.002435. [DOI] [PubMed] [Google Scholar]
  17. Schenkman S., Pontes de Carvalho L., Nussenzweig V. Trypanosoma cruzi trans-sialidase and neuraminidase activities can be mediated by the same enzymes. J Exp Med. 1992 Feb 1;175(2):567–575. doi: 10.1084/jem.175.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schmuñis G. A., Szarfman A., De Souza W., Langembach T. Trypanosoma cruzi: antibody-induced mobility of surface antigens. Exp Parasitol. 1980 Aug;50(1):90–102. doi: 10.1016/0014-4894(80)90011-9. [DOI] [PubMed] [Google Scholar]
  19. Schmuñis G. A., Szarfman A., Langembach T., de Souza W. Induction of capping in blood-stage trypomastigotes of Trypanosoma cruzi by human anti-Trypanosoma cruzi antibodies. Infect Immun. 1978 May;20(2):567–569. doi: 10.1128/iai.20.2.567-569.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Van Voorhis W. C., Barrett L., Koelling R., Farr A. G. FL-160 proteins of Trypanosoma cruzi are expressed from a multigene family and contain two distinct epitopes that mimic nervous tissues. J Exp Med. 1993 Aug 1;178(2):681–694. doi: 10.1084/jem.178.2.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Van Voorhis W. C., Schlekewy L., Trong H. L. Molecular mimicry by Trypanosoma cruzi: the F1-160 epitope that mimics mammalian nerve can be mapped to a 12-amino acid peptide. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):5993–5997. doi: 10.1073/pnas.88.14.5993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES