Role of Fas and granule exocytosis pathways in tumor-infiltrating T lymphocyte-induced apoptosis of autologous human lung-carcinoma cells

We have isolated a cytotoxic T lymphocyte (CTL) clone, Heu161, that reacts specifically with the human autologous lung carcinoma cell line IGR-Heu. We first demonstrated that IGR-Heu lacked Fas-receptor expression and was res istant to CD95-induced apoptosis. To further elucidate the role of pas in t umor immune surveillance, we have stably transfected IGR-Heu with a pas-exp ression vector and isolated CD95-sensitive and -resistant clones. Our data indicated that the resistance of 2 selected Fas-transfected clones to CD95- mediated lysis correlated with down-regulation of caspase-8 or its lack of cleavage and subsequent activation. All pas transfectants, either sensitive or resistant to anti-pas agonistic antibody, were as efficiently lysed by the CTL clone as the parental cell line. In addition, neither anti-fas-bloc king antibody nor Fas-Fc molecule inhibited T-cell lysis of Fas-sensitive t umor clone. This cytotoxicity was extracellular Ca2+-dependent and abolishe d in the presence of EGTA, indicating that it was mainly granzyme-mediated, Interestingly, although the caspase inhibitor z-VAD-fmk had no effect on t umor-cell lysis, it efficiently blocked target DNA damage triggered by auto logous CTLs via the granule exocytosis pathway, indicating that the latter event was caspase-dependent. The present results suggest that lung carcinom a-specific CTLs use mainly a granule exocytosis-dependent pathway to lyse a utologous target cells and that these effectors are able to circumvent alte ration of the pas-triggered intracellular signalling pathway via activation of a caspase-independent cytoplasmic death mechanism. (C) 2001 Wiley-Liss, Inc.