Monoclonal antibodies specific for a dimeric cell surface activation antigen (gp33/27), preliminary designated as activation inducer molecule (AIM), are capable of triggering interleukin 2 (IL 2) synthesis, IL 2 receptor expression and T cell proliferation when used in conjunction with phorbol esters. We have analyzed the functional relationship between the AIM and the CD3/TcR-mediated activation pathways. Transient modulation of the CD3/TcR complex in the Jurkat cell line, as well as the stable loss of the CD3/TcR surface expression in variant subclones, determined an inhibition of the IL 2 production triggered by anti-AIM monoclonal antibody (mAb). In contrast, neither the surface expression of AIM nor the ability to respond to a Ca2+ ionophore were affected. Similar results were observed in peripheral blood T lymphocytes, detecting after CD3 modulation an inhibition of both the IL 2 synthesis and the proliferative response to anti-AIM mAb. Altogether our data indicate that the activation pathway triggered by anti-AIM mAb is functionally linked to the expression of the CD3/TcR complex in mature T cells.