Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Nov;70(11):7773–7782. doi: 10.1128/jvi.70.11.7773-7782.1996

Identification of a gag-encoded cytotoxic T-lymphocyte epitope from FBL-3 leukemia shared by Friend, Moloney, and Rauscher murine leukemia virus-induced tumors.

W Chen 1, H Qin 1, B Chesebro 1, M A Cheever 1
PMCID: PMC190847  PMID: 8892898

Abstract

FBL-3 is a highly immunogenic murine leukemia of C57BL/6 origin induced by Friend murine leukemia virus (MuLV). Immunization of C57BL/6 mice with FBL-3 readily elicits CD8+ cytotoxic T lymphocytes (CTL) capable of lysing FBL-3 as well as syngeneic leukemias induced by Moloney and Rauscher MuLV. The aim of this current study was to identify the immunogenic epitope(s) recognized by the FBL-3-specific CD8+ CTL. A series of FBL-3-specific CD8+ CTL clones were generated from C57BL/6 mice immunized to FBL-3. The majority of CTL clones (32 of 38) were specific for F-MuLV gag-encoded antigen. By using a series of recombinant vaccinia viruses expressing full-length and truncated F-MuLV gag genes, the antigenic epitope recognized by the FBL-3 gag-specific CTL clones, as well as by bulk-cultured CTL from spleens of mice immune to FBL-3, was localized to the leader sequence of gPr80gag protein. The precise amino acid sequence of the CTL epitope in the leader sequence was identified as CCLCLTVFL (positions 85-93) by examining lysis of targets incubated with a series of synthetic leader sequence peptides. No evidence of other CTL epitopes in the gPr80gag or Pr65gag core virion structural polyproteins was found. The identity of CCLCLTVFL as the target peptide was validated by showing that immunization with the peptide elicited CTL that lysed FBL-3. The CTL elicited by the Gag peptide also specifically lysed syngeneic leukemia cells induced by Moloney and Rauscher MuLV (MBL-2 and RBL-5). The transmembrane peptide was shown to be the major gag-encoded antigenic epitope recognized by bulk-cultured CTL derived from C57BL/6 mice immunized to MBL-2 or RBL-5. Thus, the CTL epitope of FBL-3 is localized to the transmembrane anchor domain of the nonstructural Gag polyprotein and is shared by leukemia/lymphoma cell lines induced by Friend, Moloney, and Rauscher MuLV.

Full Text

The Full Text of this article is available as a PDF (258.9 KB).

Selected References

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

  1. Bacik I., Cox J. H., Anderson R., Yewdell J. W., Bennink J. R. TAP (transporter associated with antigen processing)-independent presentation of endogenously synthesized peptides is enhanced by endoplasmic reticulum insertion sequences located at the amino- but not carboxyl-terminus of the peptide. J Immunol. 1994 Jan 15;152(2):381–387. [PubMed] [Google Scholar]
  2. Berenson J. R., Einstein A. B., Jr, Fefer A. Syngeneic adoptive immunotherapy and chemoimmunotherapy of a Friend leukemia: requirement for T cells. J Immunol. 1975 Jul;115(1):234–238. [PubMed] [Google Scholar]
  3. Cheever M. A., Disis M. L., Bernhard H., Gralow J. R., Hand S. L., Huseby E. S., Qin H. L., Takahashi M., Chen W. Immunity to oncogenic proteins. Immunol Rev. 1995 Jun;145:33–59. doi: 10.1111/j.1600-065x.1995.tb00076.x. [DOI] [PubMed] [Google Scholar]
  4. Cheever M. A., Greenberg P. D., Fefer A. Specificity of adoptive chemoimmunotherapy of established syngeneic tumors. J Immunol. 1980 Aug;125(2):711–714. [PubMed] [Google Scholar]
  5. Chen W., Reese V. A., Cheever M. A. Adoptively transferred antigen-specific T cells can be grown and maintained in large numbers in vivo for extended periods of time by intermittent restimulation with specific antigen plus IL-2. J Immunol. 1990 May 15;144(10):3659–3666. [PubMed] [Google Scholar]
  6. Chesebro B., Britt W., Evans L., Wehrly K., Nishio J., Cloyd M. Characterization of monoclonal antibodies reactive with murine leukemia viruses: use in analysis of strains of friend MCF and Friend ecotropic murine leukemia virus. Virology. 1983 May;127(1):134–148. doi: 10.1016/0042-6822(83)90378-1. [DOI] [PubMed] [Google Scholar]
  7. Chesebro B., Wehrly K., Cloyd M., Britt W., Portis J., Collins J., Nishio J. Characterization of mouse monoclonal antibodies specific for Friend murine leukemia virus-induced erythroleukemia cells: friend-specific and FMR-specific antigens. Virology. 1981 Jul 15;112(1):131–144. doi: 10.1016/0042-6822(81)90619-x. [DOI] [PubMed] [Google Scholar]
  8. Chesebro B., Wehrly K. Studies on the role of the host immune response in recovery from Friend virus leukemia. II. Cell-mediated immunity. J Exp Med. 1976 Jan 1;143(1):85–99. doi: 10.1084/jem.143.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Corbin A., Prats A. C., Darlix J. L., Sitbon M. A nonstructural gag-encoded glycoprotein precursor is necessary for efficient spreading and pathogenesis of murine leukemia viruses. J Virol. 1994 Jun;68(6):3857–3867. doi: 10.1128/jvi.68.6.3857-3867.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Earl P. L., Moss B., Morrison R. P., Wehrly K., Nishio J., Chesebro B. T-lymphocyte priming and protection against Friend leukemia by vaccinia-retrovirus env gene recombinant. Science. 1986 Nov 7;234(4777):728–731. doi: 10.1126/science.3490689. [DOI] [PubMed] [Google Scholar]
  11. Falk K., Rötzschke O., Stevanović S., Jung G., Rammensee H. G. Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules. Nature. 1991 May 23;351(6324):290–296. doi: 10.1038/351290a0. [DOI] [PubMed] [Google Scholar]
  12. Feltkamp M. C., Vierboom M. P., Kast W. M., Melief C. J. Efficient MHC class I-peptide binding is required but does not ensure MHC class I-restricted immunogenicity. Mol Immunol. 1994 Dec;31(18):1391–1401. doi: 10.1016/0161-5890(94)90155-4. [DOI] [PubMed] [Google Scholar]
  13. Gilbert M. J., Riddell S. R., Li C. R., Greenberg P. D. Selective interference with class I major histocompatibility complex presentation of the major immediate-early protein following infection with human cytomegalovirus. J Virol. 1993 Jun;67(6):3461–3469. doi: 10.1128/jvi.67.6.3461-3469.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Glynn J. P., McCoy J. L., Fefer A. Cross-resistance to the transplantation of syngeneic Friend, Moloney, and Rauscher virus-induced tumors. Cancer Res. 1968 Mar;28(3):434–439. [PubMed] [Google Scholar]
  15. Greenberg P. D. Adoptive T cell therapy of tumors: mechanisms operative in the recognition and elimination of tumor cells. Adv Immunol. 1991;49:281–355. doi: 10.1016/s0065-2776(08)60778-6. [DOI] [PubMed] [Google Scholar]
  16. Hasenkrug K. J., Brooks D. M., Chesebro B. Passive immunotherapy for retroviral disease: influence of major histocompatibility complex type and T-cell responsiveness. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10492–10495. doi: 10.1073/pnas.92.23.10492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heemels M. T., Ploegh H. Generation, translocation, and presentation of MHC class I-restricted peptides. Annu Rev Biochem. 1995;64:463–491. doi: 10.1146/annurev.bi.64.070195.002335. [DOI] [PubMed] [Google Scholar]
  18. Herberman R. B., Aoki T., Nunn M., Lavrin D. H., Soares N., Gazdar A., Holden H., Chang K. S. Specificity of 51Cr-release cytotoxicity of lymphocytes immune to murine sarcoma virus. J Natl Cancer Inst. 1974 Oct;53(4):1103–1111. doi: 10.1093/jnci/53.4.1103. [DOI] [PubMed] [Google Scholar]
  19. Holt C. A., Osorio K., Lilly F. Friend virus-specific cytotoxic T lymphocytes recognize both gag and env gene-encoded specificities. J Exp Med. 1986 Jul 1;164(1):211–226. doi: 10.1084/jem.164.1.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Iwashiro M., Kondo T., Shimizu T., Yamagishi H., Takahashi K., Matsubayashi Y., Masuda T., Otaka A., Fujii N., Ishimoto A. Multiplicity of virus-encoded helper T-cell epitopes expressed on FBL-3 tumor cells. J Virol. 1993 Aug;67(8):4533–4542. doi: 10.1128/jvi.67.8.4533-4542.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Khanna R., Burrows S. R., Kurilla M. G., Jacob C. A., Misko I. S., Sculley T. B., Kieff E., Moss D. J. Localization of Epstein-Barr virus cytotoxic T cell epitopes using recombinant vaccinia: implications for vaccine development. J Exp Med. 1992 Jul 1;176(1):169–176. doi: 10.1084/jem.176.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klarnet J. P., Kern D. E., Okuno K., Holt C., Lilly F., Greenberg P. D. FBL-reactive CD8+ cytotoxic and CD4+ helper T lymphocytes recognize distinct Friend murine leukemia virus-encoded antigens. J Exp Med. 1989 Feb 1;169(2):457–467. doi: 10.1084/jem.169.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kondo T., Uenishi H., Shimizu T., Hirama T., Iwashiro M., Kuribayashi K., Tamamura H., Fujii N., Fujisawa R., Miyazawa M. A single retroviral gag precursor signal peptide recognized by FBL-3 tumor-specific cytotoxic T lymphocytes. J Virol. 1995 Nov;69(11):6735–6741. doi: 10.1128/jvi.69.11.6735-6741.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ljunggren H. G., Stam N. J., Ohlén C., Neefjes J. J., Höglund P., Heemels M. T., Bastin J., Schumacher T. N., Townsend A., Kärre K. Empty MHC class I molecules come out in the cold. Nature. 1990 Aug 2;346(6283):476–480. doi: 10.1038/346476a0. [DOI] [PubMed] [Google Scholar]
  25. Melief C. J., Kast W. M. Prospects for T cell immunotherapy of tumours by vaccination with immunodominant and subdominant peptides. Ciba Found Symp. 1994;187:97–112. doi: 10.1002/9780470514672.ch7. [DOI] [PubMed] [Google Scholar]
  26. Miyazawa M., Nishio J., Chesebro B. Protection against Friend retrovirus-induced leukemia by recombinant vaccinia viruses expressing the gag gene. J Virol. 1992 Jul;66(7):4497–4507. doi: 10.1128/jvi.66.7.4497-4507.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Murray R. J., Kurilla M. G., Brooks J. M., Thomas W. A., Rowe M., Kieff E., Rickinson A. B. Identification of target antigens for the human cytotoxic T cell response to Epstein-Barr virus (EBV): implications for the immune control of EBV-positive malignancies. J Exp Med. 1992 Jul 1;176(1):157–168. doi: 10.1084/jem.176.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Prats A. C., De Billy G., Wang P., Darlix J. L. CUG initiation codon used for the synthesis of a cell surface antigen coded by the murine leukemia virus. J Mol Biol. 1989 Jan 20;205(2):363–372. doi: 10.1016/0022-2836(89)90347-1. [DOI] [PubMed] [Google Scholar]
  29. Robertson M. N., Spangrude G. J., Hasenkrug K., Perry L., Nishio J., Wehrly K., Chesebro B. Role and specificity of T-cell subsets in spontaneous recovery from Friend virus-induced leukemia in mice. J Virol. 1992 Jun;66(6):3271–3277. doi: 10.1128/jvi.66.6.3271-3277.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ruan K. S., Lilly F. Identification of an epitope encoded in the env gene of Friend murine leukemia virus recognized by anti-Friend virus cytotoxic T lymphocytes. Virology. 1991 Mar;181(1):91–100. doi: 10.1016/0042-6822(91)90473-o. [DOI] [PubMed] [Google Scholar]
  31. Rötzschke O., Falk K., Deres K., Schild H., Norda M., Metzger J., Jung G., Rammensee H. G. Isolation and analysis of naturally processed viral peptides as recognized by cytotoxic T cells. Nature. 1990 Nov 15;348(6298):252–254. doi: 10.1038/348252a0. [DOI] [PubMed] [Google Scholar]
  32. Rötzschke O., Falk K., Stevanović S., Jung G., Walden P., Rammensee H. G. Exact prediction of a natural T cell epitope. Eur J Immunol. 1991 Nov;21(11):2891–2894. doi: 10.1002/eji.1830211136. [DOI] [PubMed] [Google Scholar]
  33. Shimizu T., Uenishi H., Teramura Y., Iwashiro M., Kuribayashi K., Tamamura H., Fujii N., Yamagishi H. Fine structure of a virus-encoded helper T-cell epitope expressed on FBL-3 tumor cells. J Virol. 1994 Dec;68(12):7704–7708. doi: 10.1128/jvi.68.12.7704-7708.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sijts A. J., Ossendorp F., Mengedé E. A., van den Elsen P. J., Melief C. J. Immunodominant mink cell focus-inducing murine leukemia virus (MuLV)-encoded CTL epitope, identified by its MHC class I-binding motif, explains MuLV-type specificity of MCF-directed cytotoxic T lymphocytes. J Immunol. 1994 Jan 1;152(1):106–116. [PubMed] [Google Scholar]
  35. Siliciano R. F., Soloski M. J. MHC class I-restricted processing of transmembrane proteins. Mechanism and biologic significance. J Immunol. 1995 Jul 1;155(1):2–5. [PubMed] [Google Scholar]
  36. Smith G. L., Murphy B. R., Moss B. Construction and characterization of an infectious vaccinia virus recombinant that expresses the influenza hemagglutinin gene and induces resistance to influenza virus infection in hamsters. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7155–7159. doi: 10.1073/pnas.80.23.7155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Townsend A., Ohlén C., Bastin J., Ljunggren H. G., Foster L., Kärre K. Association of class I major histocompatibility heavy and light chains induced by viral peptides. Nature. 1989 Aug 10;340(6233):443–448. doi: 10.1038/340443a0. [DOI] [PubMed] [Google Scholar]
  38. White H. D., Roeder D. A., Green W. R. An immunodominant Kb-restricted peptide from the p15E transmembrane protein of endogenous ecotropic murine leukemia virus (MuLV) AKR623 that restores susceptibility of a tumor line to anti-AKR/Gross MuLV cytotoxic T lymphocytes. J Virol. 1994 Feb;68(2):897–904. doi: 10.1128/jvi.68.2.897-904.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zweerink H. J., Gammon M. C., Utz U., Sauma S. Y., Harrer T., Hawkins J. C., Johnson R. P., Sirotina A., Hermes J. D., Walker B. D. Presentation of endogenous peptides to MHC class I-restricted cytotoxic T lymphocytes in transport deletion mutant T2 cells. J Immunol. 1993 Mar 1;150(5):1763–1771. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES