Homeostatic maturation programs drive human cDC2s into a tolerogenic state

• Single-cell multi-omics reveals a homeostatic maturation state in human cDC2s (DC2 hm ) • The IL7Rα + CCR7 + DC2 hm upregulate tolerance-associated machinery • DC2 hm potently induce regulatory T cell (Treg cell) differentiation • TSLP triggers homeostatic cDC2 maturation that polarizes Treg cells Single-cell multi-omics reveals a homeostatic maturation state in human cDC2s (DC2 hm ) The IL7Rα + CCR7 + DC2 hm upregulate tolerance-associated machinery DC2 hm potently induce regulatory T cell (Treg cell) differentiation TSLP triggers homeostatic cDC2 maturation that polarizes Treg cells Dendritic cells (DCs) serve as central regulators of immune activation and tolerance. While DC-mediated protective immunity is well established, the mechanisms underlying human peripheral tolerance remain unclear. Through the multi-omics analysis of human splenic DC maturation trajectories, we identified a homeostatic maturation program in type 2 conventional DCs (cDC2s). These homeostatically matured cDC2s (DC2 hm ) demonstrated the coordinated upregulation of (1) tolerance-associated transcripts (including tissue-restricted self-antigens), (2) regulatory T cell (Treg cell)-polarization machinery, and (3) the thymic stromal lymphopoietin (TSLP) receptor complex (interleukin [IL]-7Rα and TSLP receptor [TSLPR]). Functionally, IL-7Rα + CCR7 + DC2 hm exhibited potent capacity to convert naive CD4 + T cells into FOXP3 + Treg cells. TSLP stimulation recapitulated the DC2 hm phenotype in human ex vivo models and humanized mice, establishing this cytokine as a critical regulator of DC-mediated tolerance. Our findings delineate a human cDC2 homeostatic maturation program and identify DC2 hm as potential mediators of human peripheral tolerance, providing insights into autoimmune disease mechanisms and tolerance-inducing therapeutic strategies.