Resident regulatory T cells reflect the immune history of individual lymph nodes

Regulatory T cells (T_regs) are present in lymphoid and nonlymphoid tissues where they restrict immune activation, prevent autoimmunity, and regulate inflammation. T_regs in nonlymphoid tissues are typically resident, whereas those in lymph nodes (LNs) are considered to recirculate. However, T_regs in LNs are not a homogenous population, and circulation kinetics of different T_reg subsets are poorly characterized. Furthermore, whether T_regs can acquire memory T cell properties and persist for extended periods after their activation in LNs is unclear. Here, we used in situ labeling with a stabilized photoconvertible protein to uncover turnover rates of T_regs in LNs in vivo. We found that, whereas most T_regs in LNs recirculate, 10 to 20% are memory-like resident cells that remain in their respective LNs for weeks to months. Single-cell RNA sequencing revealed that LN-resident cells are a functionally and ontogenetically heterogeneous population and share the same core residency gene signature with conventional CD4⁺ and CD8⁺ T cells. Resident cells in LNs did not actively proliferate and did not require continuous T cell receptor (TCR) signaling for their residency. However, resident and circulating T_regs had distinct TCR repertoires, and each LN contained exclusive clonal subpopulations of resident T_regs. Our results demonstrate that, similar to conventional T cells, T_regs can form resident memory-like populations in LNs after adaptive immune responses. Specific and local suppression of immune responses by resident T_regs in draining LNs might provide previously unidentified therapeutic opportunities for the treatment of local chronic inflammatory conditions.