Single cell RNA sequencing and single cell TCR sequencing of Type 2 lymphocytes in human nasal polyps

We explored the heterogeneity of lymphocytes in primary human tissues with type 2 inflammation through single cell RNA sequencing. TCRs were sequenced through single cell TCR sequencing and used as barcodes to probe lineage relationships. Comparisons of transcriptional profiles were also made with CD8+ T cells and non-type 2 lymphocytes. We performed scRNA-Seq of CD4+ T cells, CD8+ T cells, and innate lymphoid cells (ILC) from human nasal polyps to define the heterogeneity of type 2 lymphocytes in diseases characterized by type 2 inflammation. We FACS sorted several subsets including: CD4+CD8-CCR4+CXCR3-CRTh2+/-CD45+ T cells; CD4-CD8+CD45+ T cells; CD8+CD4-CD45+ T cells; and ILCs (CD45+lineage-CD127+). For donors 1-3, paired samples from peripheral blood were obtained and CD45+CD4+CCR4+CRTh2+/- cells were sorted. Blood cells were hash labeled with Biolegend CD45 antibody (clone 2D1) to differentiate from polyp cells. All cells were then labeled with Biolegend CITESeq antibodies (TotalSeq C kit). For donors 4-7, cells were hash labeled with Biolegend hashtag antibodies. Cells were then loaded on the droplet-based Chromium system (10X Genomics). Single cell gene expression and TCR libraries were prepared using the 5’ 10X Genomics kit. ***Submitter declares that raw sequencing files will be submitted to dbGap due to privacy concerns.***