Treatment of adult rats with a monoclonal antibody specific for the CD5 antigen led to a dramatic reduction in the number of CD5+ cells. However, a substantial number of T cells remained as assessed by other T cell-specific antibodies. These CD5- T cells did not proliferate in response to alloantigen or mitogenic stimulation, did not generate cytotoxic T lymphocytes in vitro, and did not induce graft-vs.-host disease when injected into susceptible recipients in vivo. Re-expression of the CD5 antigen occurred when CD5- T cells were placed in an environment devoid of the anti-CD5 antibody. Re-expression of the antigen was followed by return of the T cell proliferative responses. While CD5- T cells could not proliferate in response to alloantigen they could produce interleukin 2 following a short pulse with the T cell mitogen concanavalin A. However, T cell proliferative or cytotoxic responses could not be rescued by the addition of an exogenous source of interleukin 2. We conclude that the CD5 antigen appears to be required for proliferation of resting T cells.