Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Oct 1;168(4):1211–1224. doi: 10.1084/jem.168.4.1211

Defective presentation to class I-restricted cytotoxic T lymphocytes in vaccinia-infected cells is overcome by enhanced degradation of antigen

PMCID: PMC2189091  PMID: 2459295

Abstract

Vaccinia infection interferes with the presentation of influenza Haemagglutinin (HA) and Nucleoprotein (NP) to class I-restricted CTL. The inhibitory effect is selective for certain epitopes, and is more profound during the late phase of infection. For influenza A/NT/60/68 NP, the block is present during both early and late phases of infection, and is selective for the COOH-terminal epitope defined by peptide 366-379, having no detectable effect on the presentation of the NH2-terminal epitope 50-63. The presentation of HA is inhibited only during the late phase of vaccinia infection. For both proteins, presentation is partially (NP) or completely (HA) restored by expression of rapidly degraded protein fragments in the vaccinia infected target cell. For HA, deletion of the NH2-terminal signal sequence completely overcomes the block. For NP, either a large NH2- terminal deletion or the construction of a rapidly degraded ubiquitin- NP fusion protein partially restores presentation. These results illustrate the relationship between degradation of viral proteins in the cytoplasm of an infected cell and recognition of epitopes at the cell surface by class I-restricted T cells.

Full Text

The Full Text of this article is available as a PDF (931.1 KB).

Selected References

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

  1. Bachmair A., Finley D., Varshavsky A. In vivo half-life of a protein is a function of its amino-terminal residue. Science. 1986 Oct 10;234(4773):179–186. doi: 10.1126/science.3018930. [DOI] [PubMed] [Google Scholar]
  2. Bastin J., Rothbard J., Davey J., Jones I., Townsend A. Use of synthetic peptides of influenza nucleoprotein to define epitopes recognized by class I-restricted cytotoxic T lymphocytes. J Exp Med. 1987 Jun 1;165(6):1508–1523. doi: 10.1084/jem.165.6.1508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennink J. R., Yewdell J. W., Smith G. L., Moller C., Moss B. Recombinant vaccinia virus primes and stimulates influenza haemagglutinin-specific cytotoxic T cells. Nature. 1984 Oct 11;311(5986):578–579. doi: 10.1038/311578a0. [DOI] [PubMed] [Google Scholar]
  4. Blanden R. V., Doherty P. C., Dunlop M. B., Gardner I. D., Zinkernagel R. M., David C. S. Genes required for cytotoxicity against virus-infected target cells in K and D regions of H-2 complex. Nature. 1975 Mar 20;254(5497):269–270. doi: 10.1038/254269a0. [DOI] [PubMed] [Google Scholar]
  5. Braciale T. J., Braciale V. L., Winkler M., Stroynowski I., Hood L., Sambrook J., Gething M. J. On the role of the transmembrane anchor sequence of influenza hemagglutinin in target cell recognition by class I MHC-restricted, hemagglutinin-specific cytolytic T lymphocytes. J Exp Med. 1987 Sep 1;166(3):678–692. doi: 10.1084/jem.166.3.678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carter P., Bedouelle H., Winter G. Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res. 1985 Jun 25;13(12):4431–4443. doi: 10.1093/nar/13.12.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chakrabarti S., Brechling K., Moss B. Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol. 1985 Dec;5(12):3403–3409. doi: 10.1128/mcb.5.12.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ciechanover A., Finley D., Varshavsky A. Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant ts85. Cell. 1984 May;37(1):57–66. doi: 10.1016/0092-8674(84)90300-3. [DOI] [PubMed] [Google Scholar]
  9. Clayberger C., Parham P., Rothbard J., Ludwig D. S., Schoolnik G. K., Krensky A. M. HLA-A2 peptides can regulate cytolysis by human allogeneic T lymphocytes. Nature. 1987 Dec 24;330(6150):763–765. doi: 10.1038/330763a0. [DOI] [PubMed] [Google Scholar]
  10. Coupar B. E., Andrew M. E., Both G. W., Boyle D. B. Temporal regulation of influenza hemagglutinin expression in vaccinia virus recombinants and effects on the immune response. Eur J Immunol. 1986 Dec;16(12):1479–1487. doi: 10.1002/eji.1830161203. [DOI] [PubMed] [Google Scholar]
  11. Davey J., Dimmock N. J., Colman A. Identification of the sequence responsible for the nuclear accumulation of the influenza virus nucleoprotein in Xenopus oocytes. Cell. 1985 Mar;40(3):667–675. doi: 10.1016/0092-8674(85)90215-6. [DOI] [PubMed] [Google Scholar]
  12. Gething M. J., Sambrook J. Construction of influenza haemagglutinin genes that code for intracellular and secreted forms of the protein. Nature. 1982 Dec 16;300(5893):598–603. doi: 10.1038/300598a0. [DOI] [PubMed] [Google Scholar]
  13. Gooding L. R., O'Connell K. A. Recognition by cytotoxic T lymphocytes of cells expressing fragments of the SV40 tumor antigen. J Immunol. 1983 Nov;131(5):2580–2586. [PubMed] [Google Scholar]
  14. Gotch F., Rothbard J., Howland K., Townsend A., McMichael A. Cytotoxic T lymphocytes recognize a fragment of influenza virus matrix protein in association with HLA-A2. 1987 Apr 30-May 6Nature. 326(6116):881–882. doi: 10.1038/326881a0. [DOI] [PubMed] [Google Scholar]
  15. Huddleston J. A., Brownlee G. G. The sequence of the nucleoprotein gene of human influenza A virus, strain A/NT/60/68. Nucleic Acids Res. 1982 Feb 11;10(3):1029–1038. doi: 10.1093/nar/10.3.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Koszinowski U., Gething M. J., Waterfield M. T-cell cytotoxicity in the absence of viral protein synthesis in target cells. Nature. 1977 May 12;267(5607):160–163. doi: 10.1038/267160a0. [DOI] [PubMed] [Google Scholar]
  17. Mackett M., Smith G. L., Moss B. General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol. 1984 Mar;49(3):857–864. doi: 10.1128/jvi.49.3.857-864.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marsh P. Ptac-85, an E. coli vector for expression of non-fusion proteins. Nucleic Acids Res. 1986 Apr 25;14(8):3603–3603. doi: 10.1093/nar/14.8.3603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maryanski J. L., Pala P., Corradin G., Jordan B. R., Cerottini J. C. H-2-restricted cytolytic T cells specific for HLA can recognize a synthetic HLA peptide. Nature. 1986 Dec 11;324(6097):578–579. doi: 10.1038/324578a0. [DOI] [PubMed] [Google Scholar]
  20. Pickup D. J., Ink B. S., Hu W., Ray C. A., Joklik W. K. Hemorrhage in lesions caused by cowpox virus is induced by a viral protein that is related to plasma protein inhibitors of serine proteases. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7698–7702. doi: 10.1073/pnas.83.20.7698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rougeon F., Kourilsky P., Mach B. Insertion of a rabbit beta-globin gene sequence into an E. coli plasmid. Nucleic Acids Res. 1975 Dec;2(12):2365–2378. doi: 10.1093/nar/2.12.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith G. L., Levin J. Z., Palese P., Moss B. Synthesis and cellular location of the ten influenza polypeptides individually expressed by recombinant vaccinia viruses. Virology. 1987 Oct;160(2):336–345. doi: 10.1016/0042-6822(87)90004-3. [DOI] [PubMed] [Google Scholar]
  23. St John T., Gallatin W. M., Siegelman M., Smith H. T., Fried V. A., Weissman I. L. Expression cloning of a lymphocyte homing receptor cDNA: ubiquitin is the reactive species. Science. 1986 Feb 21;231(4740):845–850. doi: 10.1126/science.3003914. [DOI] [PubMed] [Google Scholar]
  24. Taylor P. M., Davey J., Howland K., Rothbard J. B., Askonas B. A. Class I MHC molecules rather than other mouse genes dictate influenza epitope recognition by cytotoxic T cells. Immunogenetics. 1987;26(4-5):267–272. doi: 10.1007/BF00346521. [DOI] [PubMed] [Google Scholar]
  25. Tomley F., Binns M., Campbell J., Boursnell M. Sequence analysis of an 11.2 kilobase, near-terminal, BamHI fragment of fowlpox virus. J Gen Virol. 1988 May;69(Pt 5):1025–1040. doi: 10.1099/0022-1317-69-5-1025. [DOI] [PubMed] [Google Scholar]
  26. Townsend A. R., Bastin J., Gould K., Brownlee G. G. Cytotoxic T lymphocytes recognize influenza haemagglutinin that lacks a signal sequence. Nature. 1986 Dec 11;324(6097):575–577. doi: 10.1038/324575a0. [DOI] [PubMed] [Google Scholar]
  27. Townsend A. R., Gotch F. M., Davey J. Cytotoxic T cells recognize fragments of the influenza nucleoprotein. Cell. 1985 Sep;42(2):457–467. doi: 10.1016/0092-8674(85)90103-5. [DOI] [PubMed] [Google Scholar]
  28. Townsend A. R., Rothbard J., Gotch F. M., Bahadur G., Wraith D., McMichael A. J. The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell. 1986 Mar 28;44(6):959–968. doi: 10.1016/0092-8674(86)90019-x. [DOI] [PubMed] [Google Scholar]
  29. Townsend A. R., Skehel J. J. The influenza A virus nucleoprotein gene controls the induction of both subtype specific and cross-reactive cytotoxic T cells. J Exp Med. 1984 Aug 1;160(2):552–563. doi: 10.1084/jem.160.2.552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wabuke-Bunoti M. A., Taku A., Fan D. P., Kent S., Webster R. G. Cytolytic T lymphocyte and antibody responses to synthetic peptides of influenza virus hemagglutinin. J Immunol. 1984 Oct;133(4):2194–2201. [PubMed] [Google Scholar]
  31. Williams D. B., Swiedler S. J., Hart G. W. Intracellular transport of membrane glycoproteins: two closely related histocompatibility antigens differ in their rates of transit to the cell surface. J Cell Biol. 1985 Sep;101(3):725–734. doi: 10.1083/jcb.101.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yewdell J. W., Bennink J. R., Hosaka Y. Cells process exogenous proteins for recognition by cytotoxic T lymphocytes. Science. 1988 Feb 5;239(4840):637–640. doi: 10.1126/science.3257585. [DOI] [PubMed] [Google Scholar]
  33. Zweerink H. J., Askonas B. A., Millican D., Courtneidge S. A., Skehel J. J. Cytotoxic T cells to type A influenza virus; viral hemagglutinin induces A-strain specificity while infected cells confer cross-reactive cytotoxicity. Eur J Immunol. 1977 Sep;7(9):630–635. doi: 10.1002/eji.1830070910. [DOI] [PubMed] [Google Scholar]
  34. van Wyke K. L., Hinshaw V. S., Bean W. J., Jr, Webster R. G. Antigenic variation of influenza A virus nucleoprotein detected with monoclonal antibodies. J Virol. 1980 Jul;35(1):24–30. doi: 10.1128/jvi.35.1.24-30.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES