Identification of Kk-restricted T-cell epitope within influenza virus nucleoprotein

In order to determine the epitope structure in peptide NP50-63, which has been reported to be the only Kk-restricted T-cell antigen within the influenza virus (A/PR/8/34) nucleoprotein, a series of 13 peptides truncated from C- and N-termini of NP50-63 were synthesized and their sensitizing activities against Kk-restricted nucleoprotein-specific cytotoxic T lymphocytes (CTLs) were examined. One of the 13 peptides, NP50-57, sensitized L929 cells at the nM level, which was 100-1000 times lower in concentration than that at which the other peptides sensitized these cells. The presence of NP50-57 in A/PR/8/34-infected L929 cells was also investigated. Acid extracts of virus-infected cells were separated on a reverse-phase HPLC column and then anion-exchange column. By both separations, only one peak of sensitizing activity against nucleoprotein-specific CTLs was observed. The position of the peak coincided with that of the elution of NP50-57. These results strongly suggest that NP50-57 is the natural epitope in the antigenic structure, NP50-63.