Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1984 Aug 1;160(2):552–563. doi: 10.1084/jem.160.2.552

The influenza A virus nucleoprotein gene controls the induction of both subtype specific and cross-reactive cytotoxic T cells

PMCID: PMC2187454  PMID: 6206181

Abstract

Using genetically typed recombinant influenza A viruses that differ only in their genes for nucleoprotein, we have demonstrated that repeated stimulation in vitro of C57BL/6 spleen cells primed in vivo with E61-13-H17 (H3N2) virus results in the selection of a population of cytotoxic T lymphocytes (CTL) whose recognition of infected target cells maps to the gene for nucleoprotein of the 1968 virus. Influenza A viruses isolated between 1934 and 1979 fall into two groups defined by their ability to sensitize target cells for lysis by these CTL: 1934- 1943 form one group (A/PR/8/34 related) and 1946-1979 form the second group (A/HK/8/68 related). These findings complement and extend our previous results with an isolated CTL clone with specificity for the 1934 nucleoprotein (27, 28). It is also shown that the same spleen cells derived from mice primed with E61-13-H17 virus in vivo, but maintained in identical conditions by stimulation with X31 virus (which differs from the former only in the origin of its gene for NP) in vitro, results in the selection of CTL that cross-react on target cells infected with A/PR/8/1934 (H1N1) or A/Aichi/1968 (H3N2). These results show that the influenza A virus gene for NP can play a role in selecting CTL with different specificities and implicate the NP molecule as a candidate for a target structure recognized by both subtype-directed and cross-reactive influenza A-specific cytotoxic T cells.

Full Text

The Full Text of this article is available as a PDF (870.5 KB).

Selected References

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

  1. Askonas B. A., Webster R. G. Monoclonal antibodies to hemagglutinin and to H-2 inhibit the cross-reactive cytotoxic T cell populations induced by influenza. Eur J Immunol. 1980 Feb;10(2):151–156. doi: 10.1002/eji.1830100215. [DOI] [PubMed] [Google Scholar]
  2. Baez M., Palese P., Kilbourne E. D. Gene composition of high-yielding influenza vaccine strains obtained by recombination. J Infect Dis. 1980 Mar;141(3):362–365. doi: 10.1093/infdis/141.3.362. [DOI] [PubMed] [Google Scholar]
  3. Biddison W. E., Doherty P. C., Webster R. G. Antibody to influenza virus matrix protein detects a common antigen on the surface of cells infected with type A influenza viruses. J Exp Med. 1977 Sep 1;146(3):690–697. doi: 10.1084/jem.146.3.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braciale T. J., Andrew M. E., Braciale V. L. Heterogeneity and specificity of cloned lines of influenza-virus specific cytotoxic T lymphocytes. J Exp Med. 1981 Apr 1;153(4):910–923. doi: 10.1084/jem.153.4.910. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braciale T. J., Braciale V. L., Henkel T. J., Sambrook J., Gething M. J. Cytotoxic T lymphocyte recognition of the influenza hemagglutinin gene product expressed by DNA-mediated gene transfer. J Exp Med. 1984 Feb 1;159(2):341–354. doi: 10.1084/jem.159.2.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Braciale T. J. Immunologic recognition of influenza virus-infected cells. I. Generation of a virus-strain specific and a cross-reactive subpopulation of cytotoxic T cells in the response to type A influenza viruses of different subtypes. Cell Immunol. 1977 Oct;33(2):423–436. doi: 10.1016/0008-8749(77)90170-8. [DOI] [PubMed] [Google Scholar]
  7. Braciale T. J. Immunologic recognition of influenza virus-infected cells. II. Expression of influenza A matrix protein on the infected cell surface and its role in recognition by cross-reactive cytotoxic T cells. J Exp Med. 1977 Sep 1;146(3):673–689. doi: 10.1084/jem.146.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Braciale T. J. Specificity of cytotoxicity T cells directed to influenza virus hemagglutinin. J Exp Med. 1979 Apr 1;149(4):856–869. doi: 10.1084/jem.149.4.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doherty P. C., Effros R. B., Bennink J. Heterogeneity of the cytotoxic response of thymus-derived lymphocytes after immunization with influenza viruses. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1209–1213. doi: 10.1073/pnas.74.3.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Effros R. B., Doherty P. C., Gerhard W., Bennink J. Generation of both cross-reactive and virus-specific T-cell populations after immunization with serologically distinct influenza A viruses. J Exp Med. 1977 Mar 1;145(3):557–568. doi: 10.1084/jem.145.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Hackett C. J., Askonas B. A., Webster R. G., van Wyke K. Quantitation of influenza virus antigens on infected target cells and their recognition by cross-reactive cytotoxic T cells. J Exp Med. 1980 May 1;151(5):1014–1025. doi: 10.1084/jem.151.5.1014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Kees U., Krammer P. H. Most influenza A virus-specific memory cytotoxic T lymphocytes react with antigenic epitopes associated with internal virus determinants. J Exp Med. 1984 Feb 1;159(2):365–377. doi: 10.1084/jem.159.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koszinowski U. H., Allen H., Gething M. J., Waterfield M. D., Klenk H. D. Recognition of viral glycoproteins by influenza A-specific cross-reactive cytolytic T lymphocytes. J Exp Med. 1980 Apr 1;151(4):945–958. doi: 10.1084/jem.151.4.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lin Y. L., Askonas B. A. Biological properties of an influenza A virus-specific killer T cell clone. Inhibition of virus replication in vivo and induction of delayed-type hypersensitivity reactions. J Exp Med. 1981 Aug 1;154(2):225–234. doi: 10.1084/jem.154.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lubeck M. D., Palese P., Schulman J. L. Nonrandom association of parental genes in influenza A virus recombinants. Virology. 1979 May;95(1):269–274. doi: 10.1016/0042-6822(79)90430-6. [DOI] [PubMed] [Google Scholar]
  18. McMichael A. J., Askonas B. A. Influenza virus-specific cytotoxic T cells in man; induction and properties of the cytotoxic cell. Eur J Immunol. 1978 Oct;8(10):705–711. doi: 10.1002/eji.1830081007. [DOI] [PubMed] [Google Scholar]
  19. Meier-Ewert H., Compans R. W. Time course of synthesis and assembly of influenza virus proteins. J Virol. 1974 Nov;14(5):1083–1091. doi: 10.1128/jvi.14.5.1083-1091.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Murre C., Reiss C. S., Bernabeu C., Chen L. B., Burakoff S. J., Seidman J. G. Construction, expression and recognition of an H-2 molecule lacking its carboxyl terminus. Nature. 1984 Feb 2;307(5950):432–436. doi: 10.1038/307432a0. [DOI] [PubMed] [Google Scholar]
  21. Reiss C. S., Schulman J. L. Influenza type A virus M protein expression on infected cells is responsible for cross-reactive recognition by cytotoxic thymus-derived lymphocytes. Infect Immun. 1980 Aug;29(2):719–723. doi: 10.1128/iai.29.2.719-723.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. SCHULMAN J. L., KILBOURNE E. D. INDUCTION OF PARTIAL SPECIFIC HETEROTYPIC IMMUNITY IN MICE BY A SINGLE INFECTION WITH INFLUENZA A VIRUS. J Bacteriol. 1965 Jan;89:170–174. doi: 10.1128/jb.89.1.170-174.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schild G. C., Oxford J. S., Newman R. W. Evidence for antigenic variation in influenza A nucleoprotein. Virology. 1979 Mar;93(2):569–573. doi: 10.1016/0042-6822(79)90259-9. [DOI] [PubMed] [Google Scholar]
  24. Skehel J. J. Early polypeptide synthesis in influenza virus-infected cells. Virology. 1973 Nov;56(1):394–399. doi: 10.1016/0042-6822(73)90320-6. [DOI] [PubMed] [Google Scholar]
  25. Spiess P. J., Rosenberg S. A. A simplified method for the production of murine T-cell growth factor free of lectin. J Immunol Methods. 1981;42(2):213–222. doi: 10.1016/0022-1759(81)90151-4. [DOI] [PubMed] [Google Scholar]
  26. Tevethia S. S., Tevethia M. J., Lewis A. J., Reddy V. B., Weissman S. M. Biology of simian virus 40 (SV40) transplantation antigen (TrAg). IX. Analysis of TrAg in mouse cells synthesizing truncated SV40 large T antigen. Virology. 1983 Jul 30;128(2):319–330. doi: 10.1016/0042-6822(83)90259-3. [DOI] [PubMed] [Google Scholar]
  27. Townsend A. R., Skehel J. J. Influenza A specific cytotoxic T-cell clones that do not recognize viral glycoproteins. Nature. 1982 Dec 16;300(5893):655–657. doi: 10.1038/300655a0. [DOI] [PubMed] [Google Scholar]
  28. Townsend A. R., Skehel J. J., Taylor P. M., Palese P. Recognition of influenza A virus nucleoprotein by an H-2-restricted cytotoxic T-cell clone. Virology. 1984 Mar;133(2):456–459. doi: 10.1016/0042-6822(84)90413-6. [DOI] [PubMed] [Google Scholar]
  29. Townsend A. R., Taylor P. M., Mellor A. L., Askonas B. A. Recognition of Db and Kb gene products by influenza-specific cytotoxic T cells. Immunogenetics. 1983;17(3):283–294. doi: 10.1007/BF00364412. [DOI] [PubMed] [Google Scholar]
  30. Vitiello A., Sherman L. A. Recognition of influenza-infected cells by cytolytic T lymphocyte clones: determinant selection by class I restriction elements. J Immunol. 1983 Oct;131(4):1635–1640. [PubMed] [Google Scholar]
  31. Winter G., Fields S. The structure of the gene encoding the nucleoprotein of human influenza virus A/PR/8/34. Virology. 1981 Oct 30;114(2):423–428. doi: 10.1016/0042-6822(81)90223-3. [DOI] [PubMed] [Google Scholar]
  32. Yap K. L., Ada G. L., McKenzie I. F. Transfer of specific cytotoxic T lymphocytes protects mice inoculated with influenza virus. Nature. 1978 May 18;273(5659):238–239. doi: 10.1038/273238a0. [DOI] [PubMed] [Google Scholar]
  33. Yetter R. A., Barber W. H., Small P. A., Jr Heterotypic immunity to influenza in ferrets. Infect Immun. 1980 Aug;29(2):650–653. doi: 10.1128/iai.29.2.650-653.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Yewdell J. W., Frank E., Gerhard W. Expression of influenza A virus internal antigens on the surface of infected P815 cells. J Immunol. 1981 May;126(5):1814–1819. [PubMed] [Google Scholar]
  35. 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]
  36. Zweerink H. J., Courtneidge S. A., Skehel J. J., Crumpton M. J., Askonas B. A. Cytotoxic T cells kill influenza virus infected cells but do not distinguish between serologically distinct type A viruses. Nature. 1977 May 26;267(5609):354–356. doi: 10.1038/267354a0. [DOI] [PubMed] [Google Scholar]
  37. 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