A Stk4-Foxp3–NF-κB p65 transcriptional complex promotes Treg cell activation and homeostasis

The molecular programs involved in regulatory T (Treg) cell activation and homeostasis remain incompletely understood. Here, we show that T cell receptor (TCR) signaling in Treg cells induces the nuclear translocation of serine/threonine kinase 4 (Stk4), leading to the formation of an Stk4–NF-κB p65–Foxp3 complex that regulates Foxp3- and p65-dependent transcriptional programs. This complex was stabilized by Stk4-dependent phosphorylation of Foxp3 on serine-418. Stk4 deficiency in Treg cells, either alone or in combination with its homolog Stk3, precipitated a fatal autoimmune lymphoproliferative disease in mice characterized by decreased Treg cell p65 expression and nuclear translocation, impaired NF-κB p65–Foxp3 complex formation, and defective Treg cell activation. In an adoptive immunotherapy model, overexpression of p65 or the phosphomimetic Foxp3S418E in Stk3/4-deficient Treg cells ameliorated their immune regulatory defects. Our studies identify Stk4 as an essential TCR-responsive regulator of p65-Foxp3–dependent transcription that promotes Treg cell–mediated immune tolerance. Description Stk4 combines with NF-κB and Foxp3 in a TCR-regulated trimolecular complex to mediate Treg cell activation and homeostasis. Transcriptional complexes in Treg cells Studies in individuals with a deficiency in the serine/threonine kinase 4 (STK4) and in mice lacking Stk3/4 have linked this kinase to Treg cell function, and Cui et al. now describe a role of Stk4 in regulating Treg cell transcriptional programs. T cell receptor (TCR) signaling in Treg cell–induced nuclear translocation of Stk4 and formation of a trimolecular complex composed of Stk4, NF-κB, and Foxp3. Stk4 phosphorylated Foxp3 on serine-418, which facilitated stable complex formation, and this complex regulated both Foxp3- and p65-dependent transcriptional activities. Mice with Stk3/4-deficient Treg cells failed to form this complex, had defects in Treg cell activation and function, and developed a lethal autoimmune lymphoproliferative disorder. Overexpression of p65 or expression of the phosphomimetic Foxp3S418E restored the defective Treg cell function in vivo. These findings highlight a role for Stk4 in regulating Treg cell transcription.

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