Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1990 Dec;64(12):6246–6251. doi: 10.1128/jvi.64.12.6246-6251.1990

Overlapping cytotoxic T-lymphocyte and B-cell antigenic sites on the influenza virus H5 hemagglutinin.

C E Hioe 1, N Dybdahl-Sissoko 1, M Philpott 1, V S Hinshaw 1
PMCID: PMC248799  PMID: 1700833

Abstract

To define the recognition site of cytotoxic T lymphocytes (CTLs) on influenza virus H5 hemagglutinin (HA), an H5 HA-specific CTL clone was examined for the ability to recognize monoclonal antibody-selected HA variants of influenza virus A/Turkey/Ontario/7732/66 (H5N9). On the basis of 51Cr release assays with the variants, a CTL epitope was located near residue 168 of H5 HA. To define the epitope more precisely, a series of overlapping peptides corresponding to this region was synthesized and tested for CTL recognition. The minimum peptide recognized by the CTL clone encompassed residues 158 to 169 of H5 HA. Relative to the H3 HA three-dimensional structure, this CTL epitope is located near the distal tip of the HA molecule, also known as a major B-cell epitope on H3 HA. A single mutation at residue 168 (Lys to Glu) in the H5 HA variants abolished CTL recognition; this same amino acid was shown previously to be critical for B-cell recognition (M. Philpott, C. Hioe, M. Sheerar, and V. S. Hinshaw, J. Virol. 64:2941-2947, 1990). Additionally, mutations within this region of the HA molecule were associated with attenuation of the highly virulent A/Turkey/Ontario/7732/66 (H5N9) (M. Philpott, B. C. Easterday, and V.S. Hinshaw, J. Virol. 63:3453-3458, 1989). When tested for recognition of other H5 viruses, the CTL clone recognized the HA of A/Turkey/Ireland/1378/83 (H5N8) but not that of A/Chicken/Pennsylvania/1370/83 (H5N2), even though these viruses contain identical HA amino acid 158-to-169 sequences. These results suggest that differences outside the CTL epitope affected CTL recognition of the intact HA molecule. The H5 HA site defined in these studies is, therefore, important in both CTL and B-cell recognition, as well as the pathogenesis of the virus.

Full text

PDF
6246

Selected References

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

  1. Barnett B. C., Graham C. M., Burt D. S., Skehel J. J., Thomas D. B. The immune response of BALB/c mice to influenza hemagglutinin: commonality of the B cell and T cell repertoires and their relevance to antigenic drift. Eur J Immunol. 1989 Mar;19(3):515–521. doi: 10.1002/eji.1830190316. [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., Moss B. Anti-influenza virus cytotoxic T lymphocytes recognize the three viral polymerases and a nonstructural protein: responsiveness to individual viral antigens is major histocompatibility complex controlled. J Virol. 1987 Apr;61(4):1098–1102. doi: 10.1128/jvi.61.4.1098-1102.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Braciale T. J., Henkel T. J., Lukacher A., Braciale V. L. Fine specificity and antigen receptor expression among influenza virus-specific cytolytic T lymphocyte clones. J Immunol. 1986 Aug 1;137(3):995–1002. [PubMed] [Google Scholar]
  6. Braciale T. J., Sweetser M. T., Morrison L. A., Kittlesen D. J., Braciale V. L. Class I major histocompatibility complex-restricted cytolytic T lymphocytes recognize a limited number of sites on the influenza hemagglutinin. Proc Natl Acad Sci U S A. 1989 Jan;86(1):277–281. doi: 10.1073/pnas.86.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown L. E., Ffrench R. A., Gawler J. M., Jackson D. C., Dyall-Smith M. L., Anders E. M., Tregear G. W., Duncan L., Underwood P. A., White D. O. Distinct epitopes recognized by I-Ad-restricted T-cell clones within antigenic site E on influenza virus hemagglutinin. J Virol. 1988 Jan;62(1):305–312. doi: 10.1128/jvi.62.1.305-312.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Caton A. J., Brownlee G. G., Yewdell J. W., Gerhard W. The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype). Cell. 1982 Dec;31(2 Pt 1):417–427. doi: 10.1016/0092-8674(82)90135-0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Chambers T. M., Kawaoka Y., Webster R. G. Protection of chickens from lethal influenza infection by vaccinia-expressed hemagglutinin. Virology. 1988 Dec;167(2):414–421. [PubMed] [Google Scholar]
  11. De B. K., Brownlee G. G., Kendal A. P., Shaw M. W. Complete sequence of a cDNA clone of the hemagglutinin gene of influenza A/Chicken/Scotland/59 (H5N1) virus: comparison with contemporary North American and European strains. Nucleic Acids Res. 1988 May 11;16(9):4181–4182. doi: 10.1093/nar/16.9.4181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gerhard W., Hackett C., Melchers F. The recognition specificity of a murine helper T cell for hemagglutinin of influenza virus A/PR/8/34. J Immunol. 1983 May;130(5):2379–2385. [PubMed] [Google Scholar]
  13. 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]
  14. Gould K. G., Scotney H., Townsend A. R., Bastin J., Brownlee G. G. Mouse H-2k-restricted cytotoxic T cells recognize antigenic determinants in both the HA1 and HA2 subunits of the influenza A/PR/8/34 hemagglutinin. J Exp Med. 1987 Sep 1;166(3):693–701. doi: 10.1084/jem.166.3.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Graham C. M., Barnett B. C., Hartlmayr I., Burt D. S., Faulkes R., Skehel J. J., Thomas D. B. The structural requirements for class II (I-Ad)-restricted T cell recognition of influenza hemagglutinin: B cell epitopes define T cell epitopes. Eur J Immunol. 1989 Mar;19(3):523–528. doi: 10.1002/eji.1830190317. [DOI] [PubMed] [Google Scholar]
  16. Hackett C. J., Dietzschold B., Gerhard W., Ghrist B., Knorr R., Gillessen D., Melchers F. Influenza virus site recognized by a murine helper T cell specific for H1 strains. Localization to a nine amino acid sequence in the hemagglutinin molecule. J Exp Med. 1983 Aug 1;158(2):294–302. doi: 10.1084/jem.158.2.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hioe C. E., Hinshaw V. S. Induction and activity of class II-restricted, Lyt-2+ cytolytic T lymphocytes specific for the influenza H5 hemagglutinin. J Immunol. 1989 Apr 1;142(7):2482–2488. [PubMed] [Google Scholar]
  18. Hurwitz J. L., Herber-Katz E., Hackett C. J., Gerhard W. Characterization of the murine TH response to influenza virus hemagglutinin: evidence for three major specificities. J Immunol. 1984 Dec;133(6):3371–3377. [PubMed] [Google Scholar]
  19. Kawaoka Y., Nestorowicz A., Alexander D. J., Webster R. G. Molecular analyses of the hemagglutinin genes of H5 influenza viruses: origin of a virulent turkey strain. Virology. 1987 May;158(1):218–227. doi: 10.1016/0042-6822(87)90256-x. [DOI] [PubMed] [Google Scholar]
  20. Kuwano K., Braciale T. J., Ennis F. A. Cytotoxic T lymphocytes recognize a cross-reactive epitope on the transmembrane region of influenza H1 and H2 hemagglutinins. Viral Immunol. 1989 Fall;2(3):163–173. doi: 10.1089/vim.1989.2.163. [DOI] [PubMed] [Google Scholar]
  21. Lamb J. R., Eckels D. D., Lake P., Woody J. N., Green N. Human T-cell clones recognize chemically synthesized peptides of influenza haemagglutinin. Nature. 1982 Nov 4;300(5887):66–69. doi: 10.1038/300066a0. [DOI] [PubMed] [Google Scholar]
  22. Lamb J. R., Green N. Analysis of the antigen specificity of influenza haemagglutinin-immune human T lymphocyte clones: identification of an immunodominant region for T cells. Immunology. 1983 Dec;50(4):659–666. [PMC free article] [PubMed] [Google Scholar]
  23. Lukacher A. E., Braciale V. L., Braciale T. J. In vivo effector function of influenza virus-specific cytotoxic T lymphocyte clones is highly specific. J Exp Med. 1984 Sep 1;160(3):814–826. doi: 10.1084/jem.160.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morrison L. A., Braciale V. L., Braciale T. J. Expression of H-2I region-restricted cytolytic activity by an Lyt-2+ influenza virus-specific T lymphocyte clone. J Immunol. 1985 Dec;135(6):3691–3696. [PubMed] [Google Scholar]
  25. Philpott M., Easterday B. C., Hinshaw V. S. Neutralizing epitopes of the H5 hemagglutinin from a virulent avian influenza virus and their relationship to pathogenicity. J Virol. 1989 Aug;63(8):3453–3458. doi: 10.1128/jvi.63.8.3453-3458.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Philpott M., Hioe C., Sheerar M., Hinshaw V. S. Hemagglutinin mutations related to attenuation and altered cell tropism of a virulent avian influenza A virus. J Virol. 1990 Jun;64(6):2941–2947. doi: 10.1128/jvi.64.6.2941-2947.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Sette A., Buus S., Colon S., Miles C., Grey H. M. Structural analysis of peptides capable of binding to more than one Ia antigen. J Immunol. 1989 Jan 1;142(1):35–40. [PubMed] [Google Scholar]
  29. Sweetser M. T., Braciale V. L., Braciale T. J. Class I major histocompatibility complex-restricted T lymphocyte recognition of the influenza hemagglutinin. Overlap between class I cytotoxic T lymphocytes and antibody sites. J Exp Med. 1989 Oct 1;170(4):1357–1368. doi: 10.1084/jem.170.4.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Torres J. V., Wyde P. R., Atassi M. Z. Cytotoxic T lymphocyte recognition sites on influenza virus hemagglutinin. Immunol Lett. 1988 Sep;19(1):49–53. doi: 10.1016/0165-2478(88)90118-6. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Townsend A. R., McMichael A. J. Specificity of cytotoxic T lymphocytes stimulated with influenza virus. Studies in mice and humans. Prog Allergy. 1985;36:10–43. doi: 10.1159/000409860. [DOI] [PubMed] [Google Scholar]
  33. Unanue E. R. Antigen-presenting function of the macrophage. Annu Rev Immunol. 1984;2:395–428. doi: 10.1146/annurev.iy.02.040184.002143. [DOI] [PubMed] [Google Scholar]
  34. Wiley D. C., Wilson I. A., Skehel J. J. Structural identification of the antibody-binding sites of Hong Kong influenza haemagglutinin and their involvement in antigenic variation. Nature. 1981 Jan 29;289(5796):373–378. doi: 10.1038/289373a0. [DOI] [PubMed] [Google Scholar]
  35. Wilson I. A., Skehel J. J., Wiley D. C. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution. Nature. 1981 Jan 29;289(5796):366–373. doi: 10.1038/289366a0. [DOI] [PubMed] [Google Scholar]
  36. Wysocka M., Hackett C. J. Class I H-2d-restricted cytotoxic T lymphocytes recognize the neuraminidase glycoprotein of influenza virus subtype N1. J Virol. 1990 Mar;64(3):1028–1032. doi: 10.1128/jvi.64.3.1028-1032.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yewdell J. W., Bennink J. R., Smith G. L., Moss B. Influenza A virus nucleoprotein is a major target antigen for cross-reactive anti-influenza A virus cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1785–1789. doi: 10.1073/pnas.82.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES