Previous investigations of mononuclear cell antibody-dependent cell-mediated cytotoxicity (ADCC) toward tumor cells suggest that K lymphocytes and not monocytes are active in this cytotoxic reaction. This report, however, demonstrates that human monocytes are able to carry out ADCC toward three different human tumor cell lines (CEM T lymphoblasts, Raji bone marrow-derived (B) lymphoblasts, and HeLa cells). The cytolytic event was found to be temperature dependent and rapid, with most of the lysis occurring in the first 4 h of incubation. The extent of lysis was directly related to the number of monocytes (effector cells) and to the degree of antibody sensitization of the target cells. The antibody-dependent cell contact-mediated nature of the cytolytic event was confirmed by inhibition with competing nonspecific monomeric immunoglobulin and by the ability of monocytes in "innocent bystander" experiments to lyse antibody-coated targets but not nonantibody-coated target cells. Evidence that monocytes were clearly the effector cells in the monocyte preparations included the observation that preincubation of effector cells with opsonized zymosan particles abolished ADCC by monocytes, but had little effect on lymphocyte ADCC. Furthermore, no evidence for Fc receptor K lymphocyte contamination of the monocyte preparations was found using antibody-coated target cells that were selectively lysed by lymphocytes but not monocytes. We suggest that ADCC toward tumor cell targets may prove to be a useful assay of monocyte function in normal and disease states.