Single cell analysis of activated iNKT cells from murine epididymal adipose tissue and spleen

We show that iNKT cells undergo rapid and extensive transcriptional remodeling after activation with the lipid antigen αGalactosylceramide (αGalCer). A common transcriptional framework underpins the activation of heterogeneous iNKT cell populations, including NKT1, NKT2 and NKT17 cells. We show that regulatory iNKT cell populations, including iNKT cells from epididymal adipose tissue, undergo blunted activation and show reduced transcriptional remodeling after αGalCer. We find that regulatory iNKT cell populations are enriched for memory-like KLRG1+ and cMAF+ iNKT subsets, and express gene signatures associated with adaptive Tr1 cells. IL-10 producing NKT10 cells express cMAF, and show enrichment for a cMAF-associated gene network. We also show that NKT10 cells are also phenotypically similar to adaptive Tr1 cells and NKTFH cells. Unbiased single cell sequencing of sorted whole murine iNKT cells from epididymal adipose tissue (three samples) and spleen (three samples). Mice were treated with 1μg αGalCer and iNKT cells were isolated after 4 hours or 72 hours, or mice were treated with 4μg αGalCer and iNKT cells were isolated after 4 weeks without further activation, or after reactivation for 4 hours with 1μg αGalCer in vivo. This data was generated in association with the data in GSE142845.