Interleukin-2 expanded lymphocytes from lymph node and tumor biopsies of human renal cell carcinoma, breast and ovarian cancer

Adoptive immunotherapy with immune effector cells has proved to be potent f or treatment of tumors, however neither the attendant criteria for potentia l clinical efficacy of the injected cells, nor the method to prepare these cells are presently well established. Our procedure of collecting lymphocyt es from biological samples, was based on the use of low IL-2 concentrations (90 to 150 IU/ml) and on the stringent separation of lymphocytes from tumo r cells at the very early stages of their outgrowth in culture. When lympho cytes were derived from tumor biopsies (TIL), we observed differences depen ding on the histological type of tumor. In renal cell carcinoma, natural ki ller cells were expanded in 4/11 biopsies contrary to what was observed in breast cancer (92 +/- 5% of T lymphocytes from 9 biopsies). The outgrowth o f lymphocytes from breast tumors was slower and lower than from renal carci nomas. The autologous tumor cell line was more difficult to obtain from bre ast carcinoma (23%) than from renal cell carcinoma (61%) biopsies. For ovar ian cancer, short-term culture of tumor cells could be obtained for half of the tumor-invaded biological samples. Eight of the 23 tumor-derived cultur es contained more than 40% CD8 T. TIL were consistently cytolytic each time they could be evaluated. For ascitic and pleural fluids, data were of simi lar range. Tn ascitic-derived cultures, tumor cells and antigen-presenting cells are present and can be supposed to rechallenge T cells with tumor ant igens, Lymphocytes derived from lymph nodes could be expanded to a larger n umber than TIL, However, only 1/18 of these cultures contained more than 40 % CD8 T. The presence of few tumor cells in this culture was in favor of si gnificant specific and non-specific cytotoxicity in RCC lymph node cultures and higher percentages of CD8 T in breast cancer lymph nodes. Correlations could not be established between CD8 T percentages and specific in vitro c ytotoxicity in our polyclonal populations. Our conclusion is that phenotypi c and functional quality of lymphocytes is of interest when the T cells are derived 1) from tumors (RCC, breast or ovarian cancer) and isolated very e arly to avoid inhibitor factors secreted from tumor cells or 2) from lymph nodes and ascitic and pleural fluids when very few tumor cells are co-culti vated with lymphocytes at initial steps of culture. Final expansion to a nu mber of lymphocytes suitable for therapy (> 10(9)) could be attained in a s econd step of the procedure by the use of 1,000 IU/ml IL-2 each time it was assayed with 50.10(6) lymphocytes. In view of these data it appears that p henotypic and functional changes occur during culture depending on the pres ence of a particular ratio of tumor antigens, This could be artificially re produced.