Immunologic cytotoxicity against lymphocytes transformed or infected by Epstein-Barr virus (EBV) was mainly studied in an autologous in vitro system by ⁵¹Cr release assay and EBV-determined nuclear antigen (EBNA)-specific trypan blue exclusion method. When the cells of newly established EBV-transformed or spontaneously transformed lines were incubated with unfractionated autologous healthy donor lymphocytes or T-cell-depleted lymphocytes in the presence of EBV-positive autologous or allogeneic serum, the transformed cells were killed with high frequency. Exposure to lymphocytes alone or to EBV-positive serum alone was not effective. The cytotoxic reaction was directed against cells positive for EBV-induced membrane antigens (MA) but not against MA-negative transformed cells. A very small fraction (1 of 200) of healthy donor lymphocytes exposed to EBV converted into EBNA-positive and MA-positive cells, and these were also killed by the remaining autologous lymphocytes in the presence of EBV-positive serum. These results indicated that the present cytotoxic reaction represents antibody-dependent cellular cytotoxicity (ADCC), and this particular mechanism probably plays an important role in the immunologic surveillance in the protection against EBV-induced oncogenesis in seropositive individuals. Such ADCC, however, does not seem to function effectively in patients with systemic lupus erythematosus.