TGF-beta is a 25-kDa homodimeric protein that has been shown to have multiple roles in the regulation of lymphocyte activation. Previous studies have shown that TGF-beta is an inhibitor of numerous T and B lymphocyte activities. This study shows that TGF-beta is able to inhibit Th- and mitogen-induced murine B cell proliferation, as well as mitogen-induced B cell cycle entry and immunoglobulin secretion. Acridine orange analysis established that TGF-beta inhibits the LPS-induced B cell transition from G0 to G1A phase of the cell cycle. Evaluation of the effects of TGF-beta on the interactions between Th and B cells showed that TGF-beta inhibited antigen-specific Th-B cell physical interactions. Flow cytometric data showed that the ability of TGF-beta to interfere with the formation of Th-B cell conjugates was not due to decreased expression of IgD, IgM, class I, class II, LFA-1, or ICAM1 by the B cell. Taken together, these data establish that TGF-beta is able to act at multiple sites within the immune system.