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. 1993 Aug;61(8):3250–3258. doi: 10.1128/iai.61.8.3250-3258.1993

Further characterization of protective Trypanosoma cruzi-specific CD4+ T-cell clones: T helper type 1-like phenotype and reactivity with shed trypomastigote antigens.

S P Nickell 1, M Keane 1, M So 1
PMCID: PMC280996  PMID: 8335358

Abstract

We previously reported the isolation from immune mice of a panel of murine clonal T-cell lines which specifically recognize antigens expressed by the trypomastigote stage of the protozoan parasite Trypanosoma cruzi, the causative agent of human Chagas' disease. Our analysis indicated that distinct clones which recognize common as well as strain-specific antigenic determinants were represented. The immunoprotective potential of several of these T-cell clones was demonstrated by adoptive transfer of protection to naive syngeneic recipients. Here we report that these T-cell clones are all of the TH1 phenotype, as determined from their lymphokine secretion patterns. Significant levels of stimulatory activity for each clone were detected in trypomastigote supernatants, and the release of this activity was time and temperature dependent. Seven of 10 T-cell clones tested responded to nitrocellulose-immunoblotted trypomastigote proteins in the range of 90 to 47 kDa; no fewer than six distinct epitopes residing on at least five distinct polypeptide species were recognized by this panel of clones. Two clones (2G8 and 4B10) previously shown to protect in vivo responded to immunoblotted proteins in the range of 65 to 53 and 90 to 80 kD, respectively. Stimulatory activity for the latter clone was shown to be expressed on the surface of trypomastigotes and to bind specifically to wheat germ agglutinin, indicating that its target antigen is an 85-kDa trypomastigote surface glycoprotein.

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Selected References

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  1. Abou-Zeid C., Filley E., Steele J., Rook G. A. A simple new method for using antigens separated by polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from Western blots into antigen-bearing particles. J Immunol Methods. 1987 Apr 2;98(1):5–10. doi: 10.1016/0022-1759(87)90429-7. [DOI] [PubMed] [Google Scholar]
  2. Abuin G., Colli W., de Souza W., Alves M. J. A surface antigen of Trypanosoma cruzi involved in cell invasion (Tc-85) is heterogeneous in expression and molecular constitution. Mol Biochem Parasitol. 1989 Jul;35(3):229–237. doi: 10.1016/0166-6851(89)90209-0. [DOI] [PubMed] [Google Scholar]
  3. Alves M. J., Abuin G., Kuwajima V. Y., Colli W. Partial inhibition of trypomastigote entry into cultured mammalian cells by monoclonal antibodies against a surface glycoprotein of Trypanosoma cruzi. Mol Biochem Parasitol. 1986 Oct;21(1):75–82. doi: 10.1016/0166-6851(86)90081-2. [DOI] [PubMed] [Google Scholar]
  4. Andrews N. W., Hong K. S., Robbins E. S., Nussenzweig V. Stage-specific surface antigens expressed during the morphogenesis of vertebrate forms of Trypanosoma cruzi. Exp Parasitol. 1987 Dec;64(3):474–484. doi: 10.1016/0014-4894(87)90062-2. [DOI] [PubMed] [Google Scholar]
  5. Andrews N. W., Katzin A. M., Colli W. Mapping of surface glycoproteins of Trypanosoma cruzi by two-dimensional electrophoresis. A correlation with the cell invasion capacity. Eur J Biochem. 1984 May 2;140(3):599–604. doi: 10.1111/j.1432-1033.1984.tb08144.x. [DOI] [PubMed] [Google Scholar]
  6. Andrews N. W., Robbins E. S., Ley V., Hong K. S., Nussenzweig V. Developmentally regulated, phospholipase C-mediated release of the major surface glycoprotein of amastigotes of Trypanosoma cruzi. J Exp Med. 1988 Feb 1;167(2):300–314. doi: 10.1084/jem.167.2.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Araujo F. G. Development of resistance to Trypanosoma cruzi in mice depends on a viable population of L3T4+ (CD4+) T lymphocytes. Infect Immun. 1989 Jul;57(7):2246–2248. doi: 10.1128/iai.57.7.2246-2248.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Beard C. A., Wrightsman R. A., Manning J. E. Stage and strain specific expression of the tandemly repeated 90 kDa surface antigen gene family in Trypanosoma cruzi. Mol Biochem Parasitol. 1988 Apr;28(3):227–234. doi: 10.1016/0166-6851(88)90007-2. [DOI] [PubMed] [Google Scholar]
  10. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  11. Cavallesco R., Pereira M. E. Antibody to Trypanosoma cruzi neuraminidase enhances infection in vitro and identifies a subpopulation of trypomastigotes. J Immunol. 1988 Jan 15;140(2):617–625. [PubMed] [Google Scholar]
  12. Cherwinski H. M., Schumacher J. H., Brown K. D., Mosmann T. R. Two types of mouse helper T cell clone. III. Further differences in lymphokine synthesis between Th1 and Th2 clones revealed by RNA hybridization, functionally monospecific bioassays, and monoclonal antibodies. J Exp Med. 1987 Nov 1;166(5):1229–1244. doi: 10.1084/jem.166.5.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Coffman R. L., Seymour B. W., Lebman D. A., Hiraki D. D., Christiansen J. A., Shrader B., Cherwinski H. M., Savelkoul H. F., Finkelman F. D., Bond M. W. The role of helper T cell products in mouse B cell differentiation and isotype regulation. Immunol Rev. 1988 Feb;102:5–28. doi: 10.1111/j.1600-065x.1988.tb00739.x. [DOI] [PubMed] [Google Scholar]
  14. Fouts D. L., Ruef B. J., Ridley P. T., Wrightsman R. A., Peterson D. S., Manning J. E. Nucleotide sequence and transcription of a trypomastigote surface antigen gene of Trypanosoma cruzi. Mol Biochem Parasitol. 1991 Jun;46(2):189–200. doi: 10.1016/0166-6851(91)90043-6. [DOI] [PubMed] [Google Scholar]
  15. Germain R. N. Immunology. The ins and outs of antigen processing and presentation. Nature. 1986 Aug 21;322(6081):687–689. doi: 10.1038/322687a0. [DOI] [PubMed] [Google Scholar]
  16. Gonçalves M. F., Umezawa E. S., Katzin A. M., de Souza W., Alves M. J., Zingales B., Colli W. Trypanosoma cruzi: shedding of surface antigens as membrane vesicles. Exp Parasitol. 1991 Jan;72(1):43–53. doi: 10.1016/0014-4894(91)90119-h. [DOI] [PubMed] [Google Scholar]
  17. Hall B. F., Webster P., Ma A. K., Joiner K. A., Andrews N. W. Desialylation of lysosomal membrane glycoproteins by Trypanosoma cruzi: a role for the surface neuraminidase in facilitating parasite entry into the host cell cytoplasm. J Exp Med. 1992 Aug 1;176(2):313–325. doi: 10.1084/jem.176.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Harth G., Haidaris C. G., So M. Neuraminidase from Trypanosoma cruzi: analysis of enhanced expression of the enzyme in infectious forms. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8320–8324. doi: 10.1073/pnas.84.23.8320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Heinzel F. P., Sadick M. D., Holaday B. J., Coffman R. L., Locksley R. M. Reciprocal expression of interferon gamma or interleukin 4 during the resolution or progression of murine leishmaniasis. Evidence for expansion of distinct helper T cell subsets. J Exp Med. 1989 Jan 1;169(1):59–72. doi: 10.1084/jem.169.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hereld D., Krakow J. L., Bangs J. D., Hart G. W., Englund P. T. A phospholipase C from Trypanosoma brucei which selectively cleaves the glycolipid on the variant surface glycoprotein. J Biol Chem. 1986 Oct 15;261(29):13813–13819. [PubMed] [Google Scholar]
  21. Joiner K. A., daSilva W. D., Rimoldi M. T., Hammer C. H., Sher A., Kipnis T. L. Biochemical characterization of a factor produced by trypomastigotes of Trypanosoma cruzi that accelerates the decay of complement C3 convertases. J Biol Chem. 1988 Aug 15;263(23):11327–11335. [PubMed] [Google Scholar]
  22. Kahn S., Colbert T. G., Wallace J. C., Hoagland N. A., Eisen H. The major 85-kDa surface antigen of the mammalian-stage forms of Trypanosoma cruzi is a family of sialidases. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4481–4485. doi: 10.1073/pnas.88.10.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kahn S., Van Voorhis W. C., Eisen H. The major 85-kD surface antigen of the mammalian form of Trypanosoma cruzi is encoded by a large heterogeneous family of simultaneously expressed genes. J Exp Med. 1990 Aug 1;172(2):589–597. doi: 10.1084/jem.172.2.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Katzin A. M., Colli W. Lectin receptors in Trypanosoma cruzi. An N-acetyl-D-glucosamine-containing surface glycoprotein specific for the trypomastigote stage. Biochim Biophys Acta. 1983 Jan 19;727(2):403–411. doi: 10.1016/0005-2736(83)90425-x. [DOI] [PubMed] [Google Scholar]
  25. Kierszenbaum F., Pienkowski M. M. Thymus-dependent control of host defense mechanisms against Trypanosoma cruzi infection. Infect Immun. 1979 Apr;24(1):117–120. doi: 10.1128/iai.24.1.117-120.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Lima M. F., Villalta F. Host-cell attachment by Trypanosoma cruzi: identification of an adhesion molecule. Biochem Biophys Res Commun. 1988 Aug 30;155(1):256–262. doi: 10.1016/s0006-291x(88)81077-5. [DOI] [PubMed] [Google Scholar]
  28. Lima M. F., Villalta F. Trypanosoma cruzi trypomastigote clones differentially express a parasite cell adhesion molecule. Mol Biochem Parasitol. 1989 Mar 1;33(2):159–170. doi: 10.1016/0166-6851(89)90030-3. [DOI] [PubMed] [Google Scholar]
  29. Low M. G. Glycosyl-phosphatidylinositol: a versatile anchor for cell surface proteins. FASEB J. 1989 Mar;3(5):1600–1608. doi: 10.1096/fasebj.3.5.2522071. [DOI] [PubMed] [Google Scholar]
  30. Low M. G., Stiernberg J., Waneck G. L., Flavell R. A., Kincade P. W. Cell-specific heterogeneity in sensitivity of phosphatidylinositol-anchored membrane antigens to release by phospholipase C. J Immunol Methods. 1988 Oct 4;113(1):101–111. doi: 10.1016/0022-1759(88)90386-9. [DOI] [PubMed] [Google Scholar]
  31. Nogueira N., Ellis J., Chaplan S., Cohn Z. Trypanosoma cruzi: in vivo and in vitro correlation between T-cell activation and susceptibility in inbred strains of mice. Exp Parasitol. 1981 Jun;51(3):325–334. doi: 10.1016/0014-4894(81)90120-x. [DOI] [PubMed] [Google Scholar]
  32. Ouaissi M. A., Cornette J., Capron A. Identification and isolation of Trypanosoma cruzi trypomastigote cell surface protein with properties expected of a fibronectin receptor. Mol Biochem Parasitol. 1986 Jun;19(3):201–211. doi: 10.1016/0166-6851(86)90002-2. [DOI] [PubMed] [Google Scholar]
  33. Ouaissi M. A., Taibi A., Cornette J., Velge P., Marty B., Loyens M., Esteva M., Rizvi F. S., Capron A. Characterization of major surface and excretory-secretory immunogens of Trypanosoma cruzi trypomastigotes and identification of potential protective antigen. Parasitology. 1990 Feb;100(Pt 1):115–124. doi: 10.1017/s0031182000060182. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Peterson D. S., Fouts D. L., Manning J. E. The 85-kd surface antigen gene of Trypanosoma cruzi is telomeric and a member of a multigene family. EMBO J. 1989 Dec 1;8(12):3911–3916. doi: 10.1002/j.1460-2075.1989.tb08571.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  37. Plata F., Wietzerbin J., Pons F. G., Falcoff E., Eisen H. Synergistic protection by specific antibodies and interferon against infection by Trypanosoma cruzi in vitro. Eur J Immunol. 1984 Oct;14(10):930–935. doi: 10.1002/eji.1830141013. [DOI] [PubMed] [Google Scholar]
  38. Pollevick G. D., Affranchino J. L., Frasch A. C., Sánchez D. O. The complete sequence of a shed acute-phase antigen of Trypanosoma cruzi. Mol Biochem Parasitol. 1991 Aug;47(2):247–250. doi: 10.1016/0166-6851(91)90185-9. [DOI] [PubMed] [Google Scholar]
  39. Reed S. G. In vivo administration of recombinant IFN-gamma induces macrophage activation, and prevents acute disease, immune suppression, and death in experimental Trypanosoma cruzi infections. J Immunol. 1988 Jun 15;140(12):4342–4347. [PubMed] [Google Scholar]
  40. 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]
  41. Roberson E. L., Hanson W. L. Trypanosoma cruzi: effects of anti-thymocyte serum in mice and neonatal thymectomy in rats. Exp Parasitol. 1973 Oct;34(2):168–180. doi: 10.1016/0014-4894(73)90076-3. [DOI] [PubMed] [Google Scholar]
  42. Rosenberg I., Prioli R. P., Ortega-Barria E., Pereira M. E. Stage-specific phospholipase C-mediated release of Trypanosoma cruzi neuraminidase. Mol Biochem Parasitol. 1991 Jun;46(2):303–305. doi: 10.1016/0166-6851(91)90054-a. [DOI] [PubMed] [Google Scholar]
  43. Ruddle N. H., Conta B. S. Lymphotoxin and immune (gamma) interferon production by T cell lines and hybrids. Curr Top Microbiol Immunol. 1982;100:239–248. doi: 10.1007/978-3-642-68586-6_27. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. Schenkman S., Yoshida N., Cardoso de Almeida M. L. Glycophosphatidylinositol-anchored proteins in metacyclic trypomastigotes of Trypanosoma cruzi. Mol Biochem Parasitol. 1988 Jun;29(2-3):141–151. doi: 10.1016/0166-6851(88)90069-2. [DOI] [PubMed] [Google Scholar]
  46. Schmunis G. A., Cappa S. M., Traversa O. C., Janovsky J. F. The effect of immuno-depression due to neonatal thymectomy on infections with Trypanosoma cruzi in mice. Trans R Soc Trop Med Hyg. 1971;65(1):89–94. doi: 10.1016/0035-9203(71)90190-8. [DOI] [PubMed] [Google Scholar]
  47. Schreier M. H., Tees R., Nordin L., Nordin A. A. Functional characteristics of T cell hybridomas obtained by fusion of TCGF-dependent helper T cell clones with BW5147. Curr Top Microbiol Immunol. 1982;100:135–141. doi: 10.1007/978-3-642-68586-6_16. [DOI] [PubMed] [Google Scholar]
  48. Scott P., Natovitz P., Coffman R. L., Pearce E., Sher A. Immunoregulation of cutaneous leishmaniasis. T cell lines that transfer protective immunity or exacerbation belong to different T helper subsets and respond to distinct parasite antigens. J Exp Med. 1988 Nov 1;168(5):1675–1684. doi: 10.1084/jem.168.5.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Silva J. S., Morrissey P. J., Grabstein K. H., Mohler K. M., Anderson D., Reed S. G. Interleukin 10 and interferon gamma regulation of experimental Trypanosoma cruzi infection. J Exp Med. 1992 Jan 1;175(1):169–174. doi: 10.1084/jem.175.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Simon M. M., Landolfo S., Diamantstein T., Hochgeschwender U. Antigen- and lectin-sensitized murine cytolytic T lymphocyte-precursors require both interleukin 2 and endogenously produced immune (gamma) interferon for their growth and differentiation into effector cells. Curr Top Microbiol Immunol. 1986;126:173–185. doi: 10.1007/978-3-642-71152-7_21. [DOI] [PubMed] [Google Scholar]
  51. Street N. E., Mosmann T. R. Functional diversity of T lymphocytes due to secretion of different cytokine patterns. FASEB J. 1991 Feb;5(2):171–177. doi: 10.1096/fasebj.5.2.1825981. [DOI] [PubMed] [Google Scholar]
  52. Takehara H. A., Perini A., da Silva M. H., Mota I. Trypanosoma cruzi: role of different antibody classes in protection against infection in the mouse. Exp Parasitol. 1981 Aug;52(1):137–146. doi: 10.1016/0014-4894(81)90069-2. [DOI] [PubMed] [Google Scholar]
  53. Takle G. B., Cross G. A. An 85-kilodalton surface antigen gene family of Trypanosoma cruzi encodes polypeptides homologous to bacterial neuraminidases. Mol Biochem Parasitol. 1991 Oct;48(2):185–198. doi: 10.1016/0166-6851(91)90114-l. [DOI] [PubMed] [Google Scholar]
  54. Takle G. B., Young A., Snary D., Hudson L., Nicholls S. C. Cloning and expression of a trypomastigote-specific 85-kilodalton surface antigen gene from Trypanosoma cruzi. Mol Biochem Parasitol. 1989 Nov;37(1):57–64. doi: 10.1016/0166-6851(89)90102-3. [DOI] [PubMed] [Google Scholar]
  55. Tarleton R. L. Depletion of CD8+ T cells increases susceptibility and reverses vaccine-induced immunity in mice infected with Trypanosoma cruzi. J Immunol. 1990 Jan 15;144(2):717–724. [PubMed] [Google Scholar]
  56. Trischmann T. M., Bloom B. R. Trypanosoma cruzi: ability of T-cell-enriched and -depleted lymphocyte populations to passively protect mice. Exp Parasitol. 1980 Apr;49(2):225–232. doi: 10.1016/0014-4894(80)90119-8. [DOI] [PubMed] [Google Scholar]
  57. Van Voorhis W. C., Eisen H. Fl-160. A surface antigen of Trypanosoma cruzi that mimics mammalian nervous tissue. J Exp Med. 1989 Mar 1;169(3):641–652. doi: 10.1084/jem.169.3.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Velge P., Ouaissi M. A., Cornette J., Afchain D., Capron A. Identification and isolation of Trypanosoma cruzi trypomastigote collagen-binding proteins: possible role in cell-parasite interaction. Parasitology. 1988 Oct;97(Pt 2):255–268. doi: 10.1017/s0031182000058467. [DOI] [PubMed] [Google Scholar]
  59. Villalta F., Lima M. F., Howard S. A., Zhou L., Ruiz-Ruano A. Purification of a Trypanosoma cruzi trypomastigote 60-kilodalton surface glycoprotein that primes and activates murine lymphocytes. Infect Immun. 1992 Aug;60(8):3025–3032. doi: 10.1128/iai.60.8.3025-3032.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wirth J. J., Kierszenbaum F., Sonnenfeld G., Zlotnik A. Enhancing effects of gamma interferon on phagocytic cell association with and killing of Trypanosoma cruzi. Infect Immun. 1985 Jul;49(1):61–66. doi: 10.1128/iai.49.1.61-66.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Zingales B., Andrews N. W., Kuwajima V. Y., Colli W. Cell surface antigens of Trypanosoma cruzi: possible correlation with the interiorization process in mammalian cells. Mol Biochem Parasitol. 1982 Aug;6(2):111–124. doi: 10.1016/0166-6851(82)90069-x. [DOI] [PubMed] [Google Scholar]

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