Control of naive T cell reactivity and peripheral tolerance by ascorbate and TET activity

Peripheral tolerance depends on limiting conventional T cell responses to self-antigens. To define the contribution of nutritional factors and related epigenetic regulation, we perform in vivo CRISPR screening and identify the ascorbate transporter Slc23a2 as a key regulator of naive T cell (Tn) reactivity. T cell-specific loss of Slc23a2 reduces intracellular ascorbate, induces regional DNA hypermethylation, enhances differentiation of Tn cells into effector and memory populations, and promotes low-grade autoimmune inflammation. These phenotypes mirror those caused by ascorbate deprivation, TET dioxygenase deficiency, and aging. Mechanistically, Slc23a2 maintains TET enzymatic activity through ascorbate, dampens proximal TCR signaling, represses helper T cell lineage programs, and sustains Tcf1 expression and chromatin binding in Tn cells. Deletion of Tcf7 partially phenocopies Slc23a2 deficiency, whereas Tcf1 overexpression mitigates the effects. Together, these findings identify ascorbate and TET-dependent DNA demethylation as a checkpoint that restrains Tn cell reactivity to self-antigens, thereby maintaining peripheral tolerance, a process compromised with aging.