An Epstein-Barr virus (EBV)-negative lymphoblastoid cell line (LCL), BJA-B, was established from an African Burkitt's lymphoma (BL) which contained no detectable EBV DNA and did not express the EBV specified antigen EBNA, BJA-B cells grow in typically large, flat clumps. All carry surface-bound immunoglobulins, a B lymphocyte marker, and do not form rosettes with sheep erythrocytes. After infection of BJA-B cells by EBV the infected cells may produce either EBV-determined nuclear antigen (EBNA) or both EBNA and early antigen (EA), depending on the strain of EBV. The homogeneity of the BJA-B cell population with respect to immunological and isoenzyme markers and size suggests a clonal origin of the line. BJA-B is the first EBV-negative LCL established from an African Burkitt's lymphoma and demonstrates that an EBV-independent continuous B cell line can be established "in vitro" from other than leukemia or myeloma cells.