A lymphokine demonstrating human B cell differentiation factor (BCDF)-like activity was isolated from immature (MOLT-4f, CCRF-CEM and CCRF-HSDB-2) and mature (HUT-78) malignant human T lymphoid cell lines and from human B lymphoblastoid cell lines (BJAB and ALL-7031-B). All the cell lines were grown long term in serum-free medium. This BCDF-like activity has a molecular mass in the range of 40-60 kDa and stimulates immunoglobulin synthesis of cell lines capable of producing IgA (GM-1056), IgG (GM-1500 and CESS) and IgM (CBL#3). It was not produced by a myeloid cell line. We were only able to identify the differentiation activity produced by the T and B cell lines by using appropriate molecular mass fractions from the serum-free medium as controls. This BCDF-like activity is different from that of the human BCDF so far described as it has a higher molecular mass and is constitutively produced by malignant T lymphoid cell lines which are human T cell leukemia virus-I negative and by B lymphoblastoid cell lines.