To define the role of CD4 in modulating T cell signaling pathways and regulating Th1 and Th2 differentiation, we have examined the activation and differentiation characteristics of naive T cells from CD4 mutant mice. Using peptides with differing affinities for the moth cytochrome c-specific TCR, we test the hypothesis that differences in coreceptor recruitment and signaling explain the qualitatively distinct signaling pathways seen in CD4 T cells following high affinity agonist and low affinity altered peptide ligand (APL) ligation. We find that the absence of CD4 signaling during stimulation with a strong agonist peptide does not qualitatively change the pattern of early TCR-mediated biochemical signaling events into a pattern resembling the response of CD4+ T cells to APLs. In contrast, the response to APL stimulation, by T cells bearing the same TCR, does require a component of CD4 signaling. The proliferative response and calcium signals normally seen following APL stimulation are markedly diminished in the absence of CD4. In addition, we find that naive T cell differentiation into Th2 effector cells is impaired in the absence of CD4. These data suggest that the altered pattern of biochemical signals generated by APLs require CD4 coreceptor function and that some of these signals may be required to initiate Th2 differentiation.