The Regulatory Effect of Tumor Necrosis Factor and Interleukin-2 on Tumor Infiltrating Lymphocytes from Renal Cell Carcinoma Patients

Choi, Hong Yong; Lee, Tchun Yong; Woo, Young Nam

Abstract

PURPOSE: To increase the potency of TIL(tumor infiltrating lymphocytes) in immunotheraphy, the effect of TNF(tumor necrosis factor) and IL-2(interleukin-2) were investigated in vitro on proliferation and cytotoxic effect of TIL to the various kinds of tumor lines and fresh frozen tumor cells from renal cell carcinoma patient. MATERIALS AND METHOD: For the target cell of the TIL cytotoxicity was used M-l 4(Melanoma tumor line) for investigation of LAK activity and erythroleukemic cell line(K562) for investigation of natural killer(NK) cell activity. To investigate the nonspecific cytotoxicity on renal cell carcinoma, autologous fresh frozen renal carcinoma cells were used as the autologous target, and allogeneic fresh frozen renal carcinoma cells and renal cell carcinoma line(444) as the allogeneic target cells. The dosage of TNF was 500univm1 and IL-2 was 1000Cetus unit/ml. The proliferation effect of TIL was checked by {H}Thymidine uptake assay and its cytotoxicity was checked with 4 hour Chromium release cytotoxicity assay.
RESULTS
The proliferation erect of TIL, treated with IL-2 at 20ays of culture was 11486+/-591cpm and 23817+/-1069cpm with combination of IL-2 and TNF. The nonspecific cytotoxicity of TIL for the 444 at 20th day of culture was 8.2Lytic unit(LU) in IL-2 only and 18,5LU with combination of 1L-2 and TNF. The cytotoxicity of IL for M-14 at 20th day of culture was 8.2LU in IL-2 only and 19.0LU with combination of IL-2 and TNF. The cytotoxicity of TIL for the K562 at 20th day of culture was 37.3LU in IL-2 only and 60.4LU with combination of IL-2 and TNF.
CONCLUSIONS
These results suggest that the combination of TNF and IL-2 has a synergistic effect on proliferation and nonspecific cytotoxicity of TIL at 20 6ays of culture in vitro. These results bear important practical implication for clinical trials of TIL in adoptive immunotherapy.