MULTIPLE SUB-SETS OF CD4(-CELL CLONES DIRECTED TO AUTOLOGOUS HUMAN-MELANOMA IDENTIFIED BY CYTOKINES PROFILES() AND CD8(+) CYTOTOXIC T)

CD4(+) and CD8(+) cytotoxic T-cell (CTL) clones, selected for T-cell-r eceptor (TcR)-dependent lysis of the autologous tumor and isolated fro m peripheral-blood lymphocytes (PBL) or tumor-infiltrating lymphocytes (TIL) of 3 melanoma patients, were characterized for the pattern of 1 3 different cytokines release by antibody- or tumor-mediated triggerin g. Induction or enhancement of cytokine release by anti-CD3 monoclonal antibody (MAb) led to the identification of 2 major sub-sets of CD8() CTL clones on the basis of production of IL-4. Within the 2 groups o f IL-4-producing or non-producing clones, further sub-sets could be id entified on the basis of differential production of IL-1 beta, IL-2, I L-6, IL-8, IL-10, TNF-alpha, TNF beta and IFN-gamma. A similar analysi s performed on a panel of CD4(+) CTL clones indicated multiple pattern s consistent with at least 4 major sub-sets, but further complexity wa s evident in each sub-set on the basis of differential production of I L-1, IL2, IL-6, IL-10 and G-CSF. The cytokine profile of CD4(+) and CD 8(+) clones, as determined after anti-CD3 stimulation, was different f rom the pattern seen after co-culture with autologous tumor, since man y clones released cytokines such as IL-4, IL-10, IFN-alpha and -gamma, TNF-alpha and GM-CSF after activation with only 1 of the 2 stimuli. T hese results indicate that CD4(+) and CD8(+) CTL clones reacting to hu man melanoma belong to a highly complex repertoire of functional subse ts characterized by distinct cytokine profiles. In addition, the cytok ine pattern of each T-cell sub-set can be modulated by changing the ac tivation signals delivered to the T cell. (C) 1994 Wiley-Liss, Inc.