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* Department of Virology and WHO National Influenza Center, Erasmus MC, Rotterdam, The Netherlands;
Department of Clinical Viro-Immunology, Sanquin Research at Central Laboratory for Bloodtransfusion, Amsterdam, The Netherlands;
Department of Zoology, University of Cambridge, Cambridge, United Kingdom;
Santa Fe Institute, Santa Fe, NM 87501;
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Los Alamos National Laboratory, Los Alamos, NM 87545; and
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Sanquin Bloodbank Rotterdam, Laboratory for Histocompatibility and Immunogenetics, Dordrecht, The Netherlands
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.
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