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CD4 cells receiving a strong IL-2-inducing stimulus in vitro do not undergo autocrine IL-2 signaling. Naive polyclonal CD4 cells were stimulatedinvitrowithplate-boundanti-CD3 for 16 h (CD3 16 h), rested 9 h in fresh medium (rest 9 h), subsequently cultured with exogenous IL-2 (rest → IL-2) or restimulated with either PI (rest → PI) or soluble anti-CD3 plus APCs (rest → CD3) and directly stained for Foxp3, IL-2, CD25, and pSTAT5.
