Autologous Tumor-Specific Cytotoxicity of Tumor-Infiltrating Lymphocytes Derived from Human Renal Cell Carcinoma

Abstract

Conditions for generating and expanding cytotoxic tumor-specific, tumor-infiltrating lymphocytes (TIL) were studied to improve the efficacy of adoptive cancer immunotherapy. Thus, we have examined the growth and cytolytic patterns of bulk culture TIL from human renal cell carcinoma (RCC) cultured in low (20 U/ml) or high (1,000 U/ml) dose interleukin (IL)-2, with or without irradiated autologous tumor stimulation. By 55 days in culture, TIL grown in the presence of IL-2 without tumor stimulation lost their lytic activity, whereas those exposed to tumor stimulation maintained high levels of cytotoxicity against autologous and/or nonautologous tumor targets. Only TIL grown with low dose IL-2 and autologous tumor maintained long-term (over 4 months in culture) specific cytotoxicity against the autologous tumor, even upon cryopreservation and regrowth. These TIL were 88-97% and 80% positive for CD3 and CD8, with a persistent subset exhibiting CD4 + CD8 + double positive staining. Their specific cytotoxic activity was major histocompatibility complex Class I-restricted and inhibited by pretreating the TIL with anti-CD3 monoclonal antibody. TIL exposed to the four types of culture conditions, low or high dose IL-2, with or without irradiated autologous tumors, and exhibiting different lytic specificities, all expressed mRNA for interferon-gamma and tumor necrosis factor (TNF)-alpha, but not for IL-1-beta, IL-4, IL-6, and granulocyte-macrophage colony stimulating factor. The degree of TNF-alpha mRNA expression correlated with the degree of autologous tumor-specific cytotoxicity of these TIL. This initial report demonstrates that antigen-specific cytotoxicity against the autologous tumor does, in fact, exist within the RCC tumors. Furthermore, by modifying culture conditions, cytotoxic T-lymphocytes with anti-tumor activity can be selectively grown from the heterogeneous bulk cultures of TIL. Key Words: Lymphokine expression-Renal cell carcinoma-Specific immunity-Tumor-infiltrating lymphocytes.Lymphocytes in suspensions of the original tumor mass, when cultured in the presence of interleukin (IL)-2, expand and destroy the tumor cells to yield a pure population of tumor-infiltrating lym-

Flow cytometric analysis of tumour‐infiltrating lymphocytes in patients with renal cell carcinoma

“…A strong association between these factors study shows that about 70% of renal tumours have a detectable infiltrate comprising more CD8+ than CD4+ might indicate putative suppressor factors released at the tumour site that inhibit the function of infiltrating T cells. The predominance of CD3+ CD8+ cells in some RCC tumours has been reported [6,7] and agrees with lymphocytes. B cells can act as antigen-presenting cells (APC) and recent reports based on other tumour types and using diÂerent methods [8][9][10].…”

Characterization of fresh (uncultured) tumour-infiltrating lymphocytes (TIL) and TIL-derived T cell lines from patients with renal cell carcinoma

“…Several laboratories have previously developed T cell lines from RCC TIL using various concentrations of rIL-2 [5,[14][15][16][17][18][19][20]. Most of these T cell lines exhibited a wide range of cytotoxicity against autologous and allogeneic renal tumour cells and NK-sensitive targets.…”

“…Recently, Koo et ai [20] developed a RCC-TIL-derived T cell line exhibiting autologous tumour-specific cytotoxicity, using 20 Cetus units/ml (= 120IU/ml) of rIL-2 and frequent restimulation with irradiated autologous tumour cells. We report here the development ofa RCC-TIL-derived T cell line that exhibited preferential cytotoxicity against autologous tumour cells.…”

“…RCC-TIL grown in high doses of riL-2 (up to 6000U/tnl) exhibit a wide range of cytotoxicity against autologous and allogeneic targets [7,) 4-!9]. Recently, Koo et al [20] reported the development of a TIL-derived T cell line from a patient with RCC, exhibiting primarily autologous tumour-specific cytotoxicity. Both restimulation with autoiogous tumour celis and low concentrations of rIL-2 (i 20 U/ml) were critical for the development of this T celi line with autologous tumour-specific cytotoxicity.…”

Gene transfer of human interferon gamma complementary DNA into a renal cell carcinoma line enhances MHC-restricted cytotoxic T lymphocyte recognition but suppresses non-MHC-restricted effector cell activity

“…[14][15][16] More rarely, classical MHC-restricted CTL have been identified in RCC patients. [17][18][19][20][21][22][23][24] CTL lysis of autologous tumor cells is dependent upon triggering of their T cell receptors (TCR) with a ligand composed of a tumor-associated peptide complexed with an MHC molecule. 25,26 Interferon gamma (IFN␥) has several important immune modulating capacities that might contribute to the optimal development of both types of antitumor response in RCC patients.…”