The molecular events involved in the establishment and maintenance of CD4⁺ central memory and effector memory T cells (T_CM and T_EM, respectively) are poorly understood. In this study, we demonstrate that ex vivo isolated T_CM are more resistant to both spontaneous and Fas-induced apoptosis than T_EM and have an increased capacity to proliferate and persist in vitro. Using global gene expression profiling, single cell proteomics, and functional assays, we show that the survival of CD4⁺ T_CM depends, at least in part, on the activation and phosphorylation of signal transducer and activator of transcription 5a (STAT5a) and forkhead box O3a (FOXO3a). T_CM showed a significant increase in the levels of phosphorylation of STAT5a compared with T_EM in response to both IL-2 (P < 0.04) and IL-7 (P < 0.002); the latter is well known for its capacity to enhance T cell survival. Moreover, ex vivo T_CM express higher levels of the transcriptionally inactive phosphorylated forms of FOXO3a and concomitantly lower levels of the proapoptotic FOXO3a target, Bim. Experiments aimed at blocking FOXO3a phosphorylation confirmed the role of this phosphoprotein in protecting T_CM from apoptosis. Our results provide, for the first time in humans, an insight into molecular mechanisms that could be responsible for the longevity and persistence of CD4⁺ T_CM.