Single-Cell RNA Sequencing of Bone Marrow ILC2p and CHILPs, and Lung and Mesenteric Lymph Node ILC2s

To explore potential tissue origins of iILC2 during helminth infection, we performed single-cell RNA sequencing (scRNA-seq) of ILC2 and ILC progenitors using the 10x Genomics platform. Intestine draining mesenteric lymph nodes (Mes LN), lung, and bone marrow (BM) were collected 5 days post infection (dpi) with Nippostrongylus brasiliensis. Overall design: ILC2 and ILC progenitors were enriched by lineage depletion prior to cell sorting. BM cells were sorted for progenitor cell markers CD45+Lin- CD127+ a4b7+ Flt3-. These markers account for both the common helper-like ILC progenitors (CHILP) and ILC2 progenitors (ILC2p). Although CHILP are upstream of ILC2p, they were included in our sorting strategy due to their similarity to the migratory lung iILC2 population in terms of markers CD90- Arg1- ST2- a4b7+.22 Mes LN and lung cells were sorted for conventional ILC2 markers CD45+ Lin- CD127+ and KLRG1low/+.