Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Sep;64(9):3858–3862. doi: 10.1128/iai.64.9.3858-3862.1996

Epitopes recognized by human T lymphocytes in the ROP2 protein antigen of Toxoplasma gondii.

R Saavedra 1, M A Becerril 1, C Dubeaux 1, R Lippens 1, M J De Vos 1, P Hérion 1, A Bollen 1
PMCID: PMC174303  PMID: 8751939

Abstract

The ROP2 protein of Toxoplasma gondii possesses immunological and biological properties which have led to its proposal as a vaccine candidate. To identify epitopes recognized by human T cells in the ROP2 antigen, we submitted the sequence of this protein to three reported T-cell epitope prediction algorithms. Three sequences that were predicted by all three methods were selected (sequences 197 to 216, 393 to 410, and 501 to 524), and the corresponding peptides were synthesized. The peptides were first tested in a proliferation assay with a DPw4-restricted, ROP2-specific human T-cell clone, and the peptide corresponding to residues 197 to 216 was shown to stimulate the T-cell clone. The three peptides were further tested in proliferation assays with peripheral blood mononuclear cells from a panel of T. gondii-seropositive and -seronegative individuals. We found that cells from a high proportion of the seropositive donors (64%) recognized at least one of the three peptides. The most frequently recognized ones were peptides 197 to 216 (45%) and 501 to 524 (36%). None of the seronegative donors responded to any peptide. These results show that the ROP2 antigen of T. gondii contains T-cell epitopes recognized by a high percentage of the immune population and further strengthen its potential as a vaccine candidate.

Full Text

The Full Text of this article is available as a PDF (213.3 KB).

Selected References

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

  1. Beckers C. J., Dubremetz J. F., Mercereau-Puijalon O., Joiner K. A. The Toxoplasma gondii rhoptry protein ROP 2 is inserted into the parasitophorous vacuole membrane, surrounding the intracellular parasite, and is exposed to the host cell cytoplasm. J Cell Biol. 1994 Nov;127(4):947–961. doi: 10.1083/jcb.127.4.947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bugawan T. L., Horn G. T., Long C. M., Mickelson E., Hansen J. A., Ferrara G. B., Angelini G., Erlich H. A. Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DP-alpha and DP-beta loci. J Immunol. 1988 Dec 1;141(11):4024–4030. [PubMed] [Google Scholar]
  3. Chardès T., Bourguin I., Mevelec M. N., Dubremetz J. F., Bout D. Antibody responses to Toxoplasma gondii in sera, intestinal secretions, and milk from orally infected mice and characterization of target antigens. Infect Immun. 1990 May;58(5):1240–1246. doi: 10.1128/iai.58.5.1240-1246.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Claverie J. M., Kourilsky P., Langlade-Demoyen P., Chalufour-Prochnicka A., Dadaglio G., Tekaia F., Plata F., Bougueleret L. T-immunogenic peptides are constituted of rare sequence patterns. Use in the identification of T epitopes in the human immunodeficiency virus gag protein. Eur J Immunol. 1988 Oct;18(10):1547–1553. doi: 10.1002/eji.1830181012. [DOI] [PubMed] [Google Scholar]
  5. Duquesne V., Auriault C., Gras-Masse H., Boutillon C., Darcy F., Cesbron-Delauw M. F., Tartar A., Capron A. Identification of T cell epitopes within a 23-kD antigen (P24) of Toxoplasma gondii. Clin Exp Immunol. 1991 Jun;84(3):527–534. [PMC free article] [PubMed] [Google Scholar]
  6. Falk K., Rötzschke O., Stevanović S., Jung G., Rammensee H. G. Pool sequencing of natural HLA-DR, DQ, and DP ligands reveals detailed peptide motifs, constraints of processing, and general rules. Immunogenetics. 1994;39(4):230–242. doi: 10.1007/BF00188785. [DOI] [PubMed] [Google Scholar]
  7. Frenkel J. K. Adoptive immunity to intracellular infection. J Immunol. 1967 Jun;98(6):1309–1319. [PubMed] [Google Scholar]
  8. Gazzinelli R. T., Hakim F. T., Hieny S., Shearer G. M., Sher A. Synergistic role of CD4+ and CD8+ T lymphocytes in IFN-gamma production and protective immunity induced by an attenuated Toxoplasma gondii vaccine. J Immunol. 1991 Jan 1;146(1):286–292. [PubMed] [Google Scholar]
  9. Godard I., Estaquier J., Zenner L., Bossus M., Auriault C., Darcy F., Gras-Masse H., Capron A. Antigenicity and immunogenicity of P30-derived peptides in experimental models of toxoplasmosis. Mol Immunol. 1994 Dec;31(17):1353–1363. doi: 10.1016/0161-5890(94)90054-x. [DOI] [PubMed] [Google Scholar]
  10. Hammond S. A., Obah E., Stanhope P., Monell C. R., Strand M., Robbins F. M., Bias W. B., Karr R. W., Koenig S., Siliciano R. F. Characterization of a conserved T cell epitope in HIV-1 gp41 recognized by vaccine-induced human cytolytic T cells. J Immunol. 1991 Mar 1;146(5):1470–1477. [PubMed] [Google Scholar]
  11. Hughes H. P. Toxoplasmosis: the need for improved diagnostic techniques and accurate risk assessment. Curr Top Microbiol Immunol. 1985;120:105–139. doi: 10.1007/978-3-662-09197-5_6. [DOI] [PubMed] [Google Scholar]
  12. Hérion P., Hernández-Pando R., Dubremetz J. F., Saavedra R. Subcellular localization of the 54-kDa antigen of Toxoplasma gondii. J Parasitol. 1993 Apr;79(2):216–222. [PubMed] [Google Scholar]
  13. Kent S. B. Chemical synthesis of peptides and proteins. Annu Rev Biochem. 1988;57:957–989. doi: 10.1146/annurev.bi.57.070188.004521. [DOI] [PubMed] [Google Scholar]
  14. Khan I. A., Eckel M. E., Pfefferkorn E. R., Kasper L. H. Production of gamma interferon by cultured human lymphocytes stimulated with a purified membrane protein (P30) from Toxoplasma gondii. J Infect Dis. 1988 May;157(5):979–984. doi: 10.1093/infdis/157.5.979. [DOI] [PubMed] [Google Scholar]
  15. Margalit H., Spouge J. L., Cornette J. L., Cease K. B., Delisi C., Berzofsky J. A. Prediction of immunodominant helper T cell antigenic sites from the primary sequence. J Immunol. 1987 Apr 1;138(7):2213–2229. [PubMed] [Google Scholar]
  16. McCabe R., Remington J. S. Toxoplasmosis: the time has come. N Engl J Med. 1988 Feb 4;318(5):313–315. doi: 10.1056/NEJM198802043180509. [DOI] [PubMed] [Google Scholar]
  17. Nathan C. F., Prendergast T. J., Wiebe M. E., Stanley E. R., Platzer E., Remold H. G., Welte K., Rubin B. Y., Murray H. W. Activation of human macrophages. Comparison of other cytokines with interferon-gamma. J Exp Med. 1984 Aug 1;160(2):600–605. doi: 10.1084/jem.160.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rothbard J. B., Taylor W. R. A sequence pattern common to T cell epitopes. EMBO J. 1988 Jan;7(1):93–100. doi: 10.1002/j.1460-2075.1988.tb02787.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rötzschke O., Falk K. Origin, structure and motifs of naturally processed MHC class II ligands. Curr Opin Immunol. 1994 Feb;6(1):45–51. doi: 10.1016/0952-7915(94)90032-9. [DOI] [PubMed] [Google Scholar]
  20. Saavedra R., Hérion P. Human T-cell clones against Toxoplasma gondii: production of interferon-gamma, interleukin-2, and strain cross-reactivity. Parasitol Res. 1991;77(5):379–385. doi: 10.1007/BF00931632. [DOI] [PubMed] [Google Scholar]
  21. Saavedra R., de Meuter F., Decourt J. L., Hérion P. Human T cell clone identifies a potentially protective 54-kDa protein antigen of Toxoplasma gondii cloned and expressed in Escherichia coli. J Immunol. 1991 Sep 15;147(6):1975–1982. [PubMed] [Google Scholar]
  22. Sadak A., Taghy Z., Fortier B., Dubremetz J. F. Characterization of a family of rhoptry proteins of Toxoplasma gondii. Mol Biochem Parasitol. 1988 Jun;29(2-3):203–211. doi: 10.1016/0166-6851(88)90075-8. [DOI] [PubMed] [Google Scholar]
  23. Suzuki Y., Orellana M. A., Schreiber R. D., Remington J. S. Interferon-gamma: the major mediator of resistance against Toxoplasma gondii. Science. 1988 Apr 22;240(4851):516–518. doi: 10.1126/science.3128869. [DOI] [PubMed] [Google Scholar]
  24. Suzuki Y., Remington J. S. Dual regulation of resistance against Toxoplasma gondii infection by Lyt-2+ and Lyt-1+, L3T4+ T cells in mice. J Immunol. 1988 Jun 1;140(11):3943–3946. [PubMed] [Google Scholar]
  25. Velge-Roussel F., Chardès T., Mévélec P., Brillard M., Hoebeke J., Bout D. Epitopic analysis of the Toxoplasma gondii major surface antigen SAG1. Mol Biochem Parasitol. 1994 Jul;66(1):31–38. doi: 10.1016/0166-6851(94)90033-7. [DOI] [PubMed] [Google Scholar]
  26. Verhofstede C., Sabbe L., Van Renterghem L. Ability of enzyme-linked immunosorbent assays to detect early immunoglobulin G antibodies to Toxoplasma gondii. Eur J Clin Microbiol. 1987 Apr;6(2):147–151. doi: 10.1007/BF02018196. [DOI] [PubMed] [Google Scholar]
  27. Wong S. Y., Remington J. S. Toxoplasmosis in pregnancy. Clin Infect Dis. 1994 Jun;18(6):853–862. doi: 10.1093/clinids/18.6.853. [DOI] [PubMed] [Google Scholar]
  28. van Gelder P., Bosman F., de Meuter F., van Heuverswyn H., Hérion P. Serodiagnosis of toxoplasmosis by using a recombinant form of the 54-kilodalton rhoptry antigen expressed in Escherichia coli. J Clin Microbiol. 1993 Jan;31(1):9–15. doi: 10.1128/jcm.31.1.9-15.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES