FUNCTIONAL AND T-CELL RECEPTOR GENE USAGE ANALYSIS OF CYTOTOXIC T-LYMPHOCYTES IN FRESH TUMOR-INFILTRATING LYMPHOCYTES FROM HUMAN HEAD AND NECK-CANCER

Twenty-one cytotoxic T lymphocyte (CTL) clones or lines that killed au tologous tumor cells, but not allogeneic tumor, K562, or Daudi cells, were established from fresh tumor-infiltrating lymphocytes of two indi viduals (HP-1 and HP-2) with head and neck cancer by limiting dilution in the presence of recombinant interleukin-2. Sixteen (76%) of these 21 clones or lines comprised CD4(+) CTLs and the other Eve comprised C D8(+) CTLs. These observations suggest that autologous tumor cell-spec ific CD4(+) CD8(-) and CD4(-) CD8(+) CTLs are present in vivo at the t umor site in head and neck cancer. Analysis of T cell receptor (TCR) g ene arrangements in 20 of the 21 CTL isolates with reverse transcripta se and the polymerase chain reaction revealed that five of 12 and five of eight isolates from HP-1 and HP-2, respectively, were clones, the other isolates being lines comprised of two or more clones. Each CTL c lone showed a different combination of V alpha and V beta gene express ion, suggesting that more than five different tumor-associated antigen s may be expressed on head and neck cancer cells. In spite of the dive rsity of TCR alpha beta combinations, TCR V alpha 1, V alpha 3, V alph a 8, V alpha 10, V beta 8, V beta 9, and V beta 17 were also frequentl y expressed in both patients. These data suggest that specific CTLs pr oliferate oligoclonally and contribute to the specific immune response against head and neck cancer in vivo.