Autologous lymph node cell-derived tumor-specific cytotoxic T-cells for use in adoptive immunotherapy of human melanoma.

The in vitro development of tumor-specific cytotoxic T-cells from draining and tumor-involved lymph nodes obtained from melanoma patients were examined. Fresh draining or tumor-involved lymph node cells (LNC) demonstrate no significant cytotoxic activity against a variety of tumor targets including autologous melanoma. Natural killer cell (NK) activity is very low or absent in all of these specimens. Culture of the cells with irradiated autologous tumor and expansion in recombinant interleukin 2 (rIL-2) results in strong cytotoxicity for autologous tumor cells. The cultured cells are T-cells of mixed CD4 and CD8 phenotypes. Following restimulation with autologous tumor, these lines are capable of becoming specifically cytotoxic for autologous tumor as tested in direct killing and in cold target inhibition studies. The LNC yield from fresh specimens ranges from 1 X 10(7) to more than 1 X 10(9) cells averaging 5 X 10(8) cells. After the cells are cultured, we can achieve up to a 60-fold or more increase in cell numbers, that demonstrate strong cytotoxicity for melanomas. The potential for adoptive immunotherapy using such specifically sensitized cytotoxic T-cells of mixed phenotypes is discussed.