Autologous tumor-specific cytotoxic T lymphocytes in the infiltrate of human metastatic melanomas. Activation by interleukin 2 and autologous tumor cells, and involvement of the T cell receptor.

Abstract

TIL from metastatic melanoma proliferated by greater than 1,000-fold (840-3,675, mean 1,543) after 6 wk in culture of mixtures of TIL and tumor cells with rIL-2 alone. Cytolysis was restricted to autologous tumor cells. CD8+ T cells were the predominant population of TIL before and after expansion, and were primarily responsible for autologous tumor-specific CTL activity. No other rIL-2-activated lymphocytes from peripheral blood, lymph nodes with melanoma metastasis, or TIL from sarcoma or renal cell carcinoma had autologous tumor-specific CTL activity. There were few or no CD16+ NK cells in TIL from metastatic melanoma before or after incubation with rIL-2, respectively. However, TIL from sarcoma or renal cell carcinoma contained a substantial proportion of CD3-CD16+ NK cells, which increased in number in culture with rIL-2. Purified CD16+ NK cells as well as CD3+CD16- T cells from rIL-2-activated TIL of renal cell carcinoma displayed MHC-nonrestricted cytotoxicity. At the clonal level as determined by limiting dilution, 8 of 10 clones from melanoma TIL displayed cytotoxicity restricted to autologous tumor cells, while all 13 clones from renal cancer TIL equally lysed autologous and allogeneic tumor cells. Anti-T cell receptor (TCR)-alpha/beta(WT31) mAb as well as anti-CD3 mAb inhibited autologous melanoma cell-specific CTL activity mediated by rIL-2-activated TIL at the effector phase. These two mAbs also inhibited rIL-2-dependent proliferation of these TIL when added to the culture. Pretreatment of fresh melanoma cells with mAb to MHC antigens followed by washing inhibited specific CTL activity. These results suggest that both TCR-alpha/beta on effector TIL and MHC antigens on fresh tumor cells are involved in the specific immune-recognition. After reaching maximum propagation, TIL from metastatic melanoma responded poorly to rIL-2 alone. However, stimulation with fresh autologous melanoma cells restored both CTL activity and proliferation in response to rIL-2. The latter is associated with IL-2 receptor (Tac antigen) expression on the surface. These results indicate that TIL from metastatic melanomas may have unique characteristics different from lymphocytes obtained from the other sources, and may contain precursor CTL sensitized in vivo to autologous tumor cells, and thus can be propagated in larger numbers with rIL-2 alone while retaining autologous tumor-specific CTL activity.

Limited heterogeneity in the T-cell receptor V-gene usage in lymphocytes infiltrating human colorectal tumours

“…Several groups have demonstrated autologous tumour-reactive, MHC-restricted tumourinfiltrating T lymphocytes (TILs) derived from malignant melanomas (Herin et al, 1987;Itoh et al, 1988), and similar results have been obtained with TILs from head and neck cancers (Yasumura et al, 1993) and ovarian carcinomas (Ioannides et al, 1993). In vivo, TILs appear to be more effective in adoptive immunotherapy than autologous peripheral blood lymphocytes (PBLs) (Rosenberg et al, 1988), suggestive of a primed population in the tumour area.…”

“…However, most studies so far have focused on the functional testing of T-cell cytotoxicity (Herin et al, 1987;Itoh et al, 1988;Yasumura et al, 1993) or cytokine secretion (Belldegrun et al, 1989;Hom et al, 1993 The fact that different sets of families predominate in each tumour therefore argue against a biased amplification due to variable quality of the primers. The inter-sample variability may actually serve as internal controls.…”

Localization of T cell receptor (TCR)- γδ+ T cells into human colorectal cancer: flow cytometric analysis of TCR-γδ expression in tumour-infiltrating lymphocytes

“…There is no doubt that tumour-specific T cells exist in TIL [17,18]. However, most lymphocytes in TIL seem not to l?e directed against autologous tumour cells, in as much as tumour-specific cytotoxic lymphocytes are difiicult to detect.…”

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

“…They did not lyse or respond to allogeneic tumour cells or K562 cells. They were of the CD3^CD8' phenotype in ovarian carcinoma [8,9,[26][27][28] and cither CD3^CD4'^CD8" or CD3'CD4"CD8' phenotype in melanoma [5,7,25]. Blocking experiments using MoAb recognizing the CD3 antigen or the a/^TCR inhibited cytotoxicity at the effector cell level, demonstrating that these cell surface structures were involved in the cytolytic process.…”

“…The intense presence of TIL has been associated in certain tumours with favourable prognosis and increased survival [2][3][4]. TIL from patients with malignant melanoma [5][6][7] or ovarian carcinoma [8,9] can be expanded in culture with recombinant IL-2 (rIL-2) over 1500-fold, and often result in T cell lines exhibiting primarily autologous tumour-specific cytotoxicity [5][6][7][8][9].…”