Cells from the murine B lymphoma I.29, expressing IgM or IgA of identical idiotype, were found inducible by lipopolysaccharide to differentiate into plasma cells. Within 3 days, differentiating cells lost membrane-bound immunoglobulin (Ig) and accumulated large quantities of intracytoplasmic Ig. At day 6 of culture, IgA secretion increased 50-100-fold, as determined by enzyme-linked immunoassay. Proliferation increased for the first days of culture but decreased thereafter; by day 10 very few viable cells were present in lipopolysaccharide-stimulated cultures. Similar results were obtained by culturing I.29 cells in the presence of supernatants of certain B cell lines (e.g. BFO.3). The finding of a strict correlation between the inductive activity and presence of contaminating Mycoplasma fermentans suggested that factor(s) released by mycoplasma were responsible for the mitogenic activities. This was further indicated by the findings that: the supernatants of BFO.3 that were rendered free of mycoplasma were not inductive, and a nonactive cell line could be made active by infection with supernatants of BFO.3 cells containing viable microorganisms. Thus, supernatants of mycoplasma-infected cell lines may act as potent polyclonal activators on both normal and malignant B lymphocytes. The ability to induce membrane Ig on 70Z/3 cells indicates that mycoplasma-related mitogens are also active on pre-B cells. The possibility of mycoplasma contamination should thus be carefully excluded when presumptive factors of cloned cell lines are being evaluated.