EBV-transformed B lymphoblastoid cell lines and clones were cytotoxic in vitro against Actinomycin D-pretreated WEHI 164 sarcoma cells, a system in which mononuclear phagocytes were previously identified as effectors. Similarly, unlike resting B cells, normal B lymphoblasts stimulated for 72 hr with Staphylococcus aureus Cowan I and purified according to the expression of the B cell surface marker B1, demonstrated appreciable cytolytic activity. B lymphoblastoid cells and derived supernatants were cytotoxic for Actinomycin D-pretreated WEHI 164 cells, and killing was inhibited by an anti-lymphotoxin but not an anti-tumor necrosis factor antiserum. Thus, B lymphoblasts have cytotoxic potential, mediated by lymphotoxin or a lymphotoxin-like soluble product.