Induction of a colitogenic phenotype in Th1-like cells depends on interleukin-23 receptor signaling [in vitro]

Single-cell RNA-sequencing of in vitro differentiated T cells cultured with IL-12+IL-21+IL-23 (Th1 cell condition) or IL-1b+IL-6+IL-23 (pathogenic Th17 cell condition) Naive T cells were isolated from reporter mice for IL-23R that carry an IRES-eGFP in the Il23r locus. Homozygous mice carrying the reporter allele (Il23reGFP/eGFP) represent a functional KO mouse. Naive T cells (CD44lowCD62LhiCD4+CD25-) were isolated from heterozygous mice (Il23rwt/eGFP=ctrl) and homozygous mice (Il23reGFP/eGFP=KO) by fluoresence-activated cell sorting (FACS) and then cultured for 4 days with full medium containing the aforementioned cytokine conditions. Cells were then sorted based on eGFP expression before single-cell sequencing.