Constitutive androstane receptor directs developing Th1 cells towards the Tr1 lineage

Abstract

Constitutive androstane receptor (CAR; encoded by Nr1i3) is a nuclear xenobiotic receptor that mediates hepatic drug and bile acid metabolism. We previously identified CAR as also operating in CD4+ T helper (TH) cells, where CAR-dependent gene expression mitigates bile acid toxicity and promotes a Foxp3-IL-10+type 1 regulatory (Tr1)-like phenotype in the small intestine. Here, we show that CAR acts early and specifically during the priming of type 1 immune responses to stabilize Tr1 lineage commitment. CAR-dependent Tr1 cells formed during type 1 (Th1-associated) but not type 3 (Th17-associated) intestinal inflammation. In vitro, IL-27 upregulated CAR expression during naive TH cell activation, which contributed to Il10 induction. Single-cell analysis revealed that naive TH cells primed with IL-27 adopt a multipotent "Th1/Tr1 precursor" (THR1p) transcriptional state, which subsequently diverges into Th1 or Tr1 developmental trajectories. CAR transcriptional activity peaked in THR1p cells, upregulating Tr1 genes, including Il10, and repressing Th1 genes. Moreover, glucocorticoid receptor activation--which increases CAR expression in hepatocytes--synergized with IL-27 to augment both CAR expression and CAR-dependent Tr1 differentiation. Together, these results suggest that CAR acts in a lineage-biased manner to enforce Tr1-mediated immune tolerance during type 1 intestinal inflammation, and this pathway is amplified by glucocorticoids.