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The infection of a number of new and established B‐cell lines by human T‐cell lymphotropic virus III (HTLV‐III) was investigated. The B lymphocytes differed in their expression of T4 antigens detected by specific monoclonal antibodies (MAbs) and the presence of Epstein Barr virus (EBV)‐DNA or antigens. The presence of the EBV genome was the only requirement for infection of B‐lymphocytes by HTLV‐III, although its presence did not ensure infection. Two EBV genome and T4 antigen‐positive B‐cell lines, lacking EBV early antigens (EA) and viral capsid antigens (VCA), could be productively infected with no induction of known EBV antigens. Two other EBV genome‐positive cell lines, lacking T4, EA, and VCA could also be infected. Another genome‐positive cell line (P3HR‐I) that was EBV‐EA, VCA‐positive and produced non‐transforming EBV, could also be infected by HTLV‐III. However, 3 EBV genome …