Recognition of Homo- and Heterosubtypic Variants of Influenza A Viruses by Human CD8+ T Lymphocytes1

In the present study, the recognition of epitope variants of influenza A viruses by human CTL was investigated. To this end, human CD8+ CTL clones, specific for natural variants of the HLA-B*3501-restricted epitope in the nucleoprotein (NP418–426), were generated. As determined in 51Cr release assays and by flow cytometry with HLA-B*3501-peptide tetrameric complexes, CTL clones were found to be specific for epitopes within one subtype or cross-reactive with heterosubtypic variants of the epitope. Using eight natural variants of the epitope, positions in the 9-mer important for T cell recognition and involved in escape from CTL immunity were identified and visualized using multidimensional scaling. It was shown that positions 4 and 5 in the 9-mer epitope were important determinants of T cell specificity. The in vivo existence of CD8+ cells cross-reactive with homo- and heterosubtypic variants of the epitope was further confirmed using polyclonal T cell populations obtained after stimulation of PBMC with different influenza A viruses. Based on the observed recognition patterns of the clonal and polyclonal T cell populations and serology, it is hypothesized that consecutive infections with influenza viruses containing different variants of the epitope select for cross-reactive T cells in vivo.

[1]  R Farber,et al.  The geometry of shape space: application to influenza. , 2001, Journal of theoretical biology.

[2]  J. Skehel,et al.  Cytotoxic T cells kill influenza virus infected cells but do not distinguish between serologically distinct type A viruses , 1977, Nature.

[3]  J Goodman,et al.  The value of a database in surveillance and vaccine selection , 2001 .

[4]  J. Ulmer,et al.  Heterologous protection against influenza by injection of DNA encoding a viral protein. , 1993, Science.

[5]  A. Osterhaus,et al.  A randomized, double blind study in young healthy adults comparing cell mediated and humoral immune responses induced by influenza ISCOM vaccines and conventional vaccines. , 2000, Vaccine.

[6]  G L Ada,et al.  Options for the control of influenza III. Cairns, North Queensland, Australia (4-9 May 1996). , 1997, Vaccine.

[7]  J. Woody,et al.  Antigen-specific human T lymphocyte clones: viral antigen specificity of influenza virus-immune clones. , 1982, Journal of immunology.

[8]  G. Oster,et al.  Theoretical studies of clonal selection: minimal antibody repertoire size and reliability of self-non-self discrimination. , 1979, Journal of theoretical biology.

[9]  B. Walker,et al.  Cytotoxic T-lymphocyte cross-reactivity among different human immunodeficiency virus type 1 clades: implications for vaccine development , 1997, Journal of virology.

[10]  A. Osterhaus,et al.  The Magnitude and Specificity of Influenza A Virus-Specific Cytotoxic T-Lymphocyte Responses in Humans Is Related to HLA-A and -B Phenotype , 2002, Journal of Virology.

[11]  T. Schumacher,et al.  Selective Expansion of Cross-Reactive Cd8+ Memory T Cells by Viral Variants , 1999, The Journal of experimental medicine.

[12]  F. Ennis,et al.  Human CD8+ and CD4+ T lymphocyte memory to influenza A viruses of swine and avian species. , 1999, Journal of immunology.

[13]  T. Croghan,et al.  Transgenic mice lacking class I major histocompatibility complex- restricted T cells have delayed viral clearance and increased mortality after influenza virus challenge , 1992, The Journal of experimental medicine.

[14]  A. Osterhaus,et al.  Antigenic Drift in the Influenza A Virus (H3N2) Nucleoprotein and Escape from Recognition by Cytotoxic T Lymphocytes , 2000, Journal of Virology.

[15]  J. Berzofsky,et al.  Selective expansion of high- or low-avidity cytotoxic T lymphocytes and efficacy for adoptive immunotherapy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[16]  C. Rouzioux,et al.  Patient-Specific Cytotoxic T-Lymphocyte Cross-Recognition of Naturally Occurring Variants of a Human Immunodeficiency Virus Type 1 (HIV-1) p24gagEpitope by HIV-1-Infected Children , 2001, Journal of Virology.

[17]  D. Baarle,et al.  Dysfunctional Epstein-Barr virus (EBV)-specific CD8(+) T lymphocytes and increased EBV load in HIV-1 infected individuals progressing to AIDS-related non-Hodgkin lymphoma. , 2001, Blood.

[18]  T. Braciale,et al.  Heterogeneity and specificity of cloned lines of influenza-virus specific cytotoxic T lymphocytes , 1981, The Journal of experimental medicine.

[19]  F. Ennis,et al.  Recognition of disparate HA and NS1 peptides by an H-2Kd-restricted, influenza specific CTL clone. , 1991, Molecular immunology.

[20]  A. Osterhaus,et al.  Sequence Variation in a Newly Identified HLA-B35-Restricted Epitope in the Influenza A Virus Nucleoprotein Associated with Escape from Cytotoxic T Lymphocytes , 2002, Journal of Virology.

[21]  G. Ada,et al.  Transfer of specific cytotoxic T lymphocytes protects mice inoculated with influenza virus , 1978, Nature.

[22]  G. M. Cross.  Hemagglutination inhibition assays , 2002 .

[23]  Philip J. R. Goulder,et al.  Phenotypic Analysis of Antigen-Specific T Lymphocytes , 1996, Science.

[24]  J. Bennink,et al.  Generation of both cross-reactive and virus-specific T-cell populations after immunization with serologically distinct influenza A viruses , 1977, The Journal of experimental medicine.

[25]  R N SHEPARD,et al.  Analysis of Proximities as a Technique for the Study of Information Processing in Man1 , 1963, Human factors.

[26]  R. Welsh,et al.  T cell immunodominance and maintenance of memory regulated by unexpectedly cross-reactive pathogens , 2002, Nature Immunology.

[27]  N. Masurel,et al.  Antibody response to immunization with influenza A/USSR/77 (H1N1) virus in young individuals primed or unprimed for A/New Jersey/76 (H1N1) virus , 1981, Journal of Hygiene.

[28]  J. Kruskal.  Multidimensional scaling by optimizing goodness of fit to a nonmetric hypothesis , 1964 .

[29]  J. Skehel,et al.  Cytotoxic T cells to type A influenza virus; viral hemagglutinin induces A‐strain specificity while infected cells confer cross‐reactive cytotoxicity , 1977, European journal of immunology.

[30]  A. Osterhaus,et al.  Comparison of RNA hybridization, hemagglutination assay, titration of infectious virus and immunofluorescence as methods for monitoring influenza virus replication in vitro. , 1998, Journal of virological methods.

[31]  F. Ennis,et al.  Cross-reactive protection against influenza A virus infections by an NS1-specific CTL clone. , 1990, Virology.

[32]  A. McMichael,et al.  Cytotoxic T-cell immunity to influenza. , 1983, The New England journal of medicine.

[33]  B. Askonas,et al.  Rapid recovery of lung histology correlates with clearance of influenza virus by specific CD8+ cytotoxic T cells. , 1989, Immunology.

[34]  F. Ennis,et al.  A single nine-amino acid peptide induces virus-specific, CD8+ human cytotoxic T lymphocyte clones of heterogeneous serotype specificities , 1995, The Journal of experimental medicine.

[35]  J. Ulmer,et al.  Dose dependence of CTL precursor frequency induced by a DNA vaccine and correlation with protective immunity against influenza virus challenge. , 1999, Journal of immunology.

[36]  A. Fraire,et al.  Memory CD8+ T cells in heterologous antiviral immunity and immunopathology in the lung , 2001, Nature Immunology.