Interleukin-6 (IL-6) is a pleiotropic cytokine with multiple immunomodulatory functions. Although IL-6 enhances cytotoxic effector cell function in vitro, we report the paradoxical effect of IL-6-induced resistance of target cells to lysis by cytotoxic T lymphocytes (CTL). The CTL system employed autologous, Epstein-Barr virus-transformed B lymphoblastoid target cells infected with vaccinia virus vectors carrying the envelope gene from the human immunodeficiency virus (HIV). Effector cells were fresh peripheral blood mononuclear cells from HIV+ individuals. Resistance was induced by exposing B cell line targets to exogenous IL-6, or via an autocrine pathway in which IL-6 was secreted by the target cells themselves. The IL-6 effect was dose dependent and reversible by antibody to IL-6. A large proportion of B cell lines from HIV+ individuals produced IL-6, and the lysis of HIV envelope-expressing B cell targets was inversely proportional to the amounts of IL-6 produced by the cell lines. These findings have significance for the utility and interpretation of CTL assays as in vitro correlates of T cell competence and may be significant in vivo in situations such as HIV infection where IL-6 production is increased.