T lymphocytes can be induced to produce interleukin (IL)-2 and proliferate upon T cell receptor (TcR) occupancy together with a CD28-induced co-stimulatory signal. The T cell surface molecule CD28 is believed to function as a regulator in T cell activation at both the level of lymphokine mRNA stabilization and gene transcription. Activation of IL-2 gene transcription via CD28 has been shown to be mediated through a kappa B-like sequence, called CD28RE. DNA binding analysis revealed that the CD28-induced signal is involved in the induction of CD28RE binding activity. Here, we demonstrate that the induction of CD28RE binding activity is not specific for the CD28-induced signal. Our data indicate that distinct mitogenic T cell activation signals converge on the induction of CD28RE binding activity, and suggest a crucial role for this activity in the IL-2 enhancer responsiveness to different modes of T cell activation.