Single cell analysis of CD4 T cells and an ILCs in nasal polyps

Chronic rhinosinusitis with nasal polyps (CRSwNP) is characterized by a chronic inflammatory process often associated with comorbid asthma. In this study, we analysed the transcriptomes of single T helper (Th) cells from nasal polyps of patients with CRS and validated these findings using multiparameter flow cytometry. Polyp tissue contained suppressive Treg and Th2 cells, type 2 innate lymphoid cells (ILC2) and 3 transcriptionally distinct subsets of cytotoxic CD4 T cells (CD4 CTL). GATA3 expression was a feature of polyp Treg while Th2 cells highly expressed TCN1, CD200R, HPGDS and were enriched for genes involved in lipid metabolism. Only a portion of polyp Th2 cells expressed the prostaglandin D2 receptor CRTH2, while a subpopulation of CD109+CRTH2- Th2 cells expressed mRNA for common inhibitor receptors and produced IL-10. Taken together, we resolve the complexity of T helper cells in CRSwNP patients to identify several distinct clusters of CD4 CTL and a population of CD109+CRTH2- Th2 cells with putative regulatory potential. Herein, we performed scRNA-Seq of CD4 T cells and functionally related innate lymphoid cells  (ILC) in nasal polyp tissue, with a view to defining the functional heterogeneity of these cells in CRSwNP patients. Viable Th cells (CD4+ CD3+ CD20- CD14- ) and ILC2 (CD4- CD3- CD20- CD14- CRTH2+ ) were sorted using an Aria FUSION sorter using FACS DIVA software (BD Biosciences). Cells were then washed in cold PBS prior to loading on the droplet-based Chromium microfluidic system (10X Genomics). Paired gene expression and TCR libraries were prepared using the 5’ v1 10X Genomics kit. ------------------------- >>>Submitter declares that the raw data for samples will be deposited in EGA due to patient privacy concerns.<<<