Identification of M3 muscarinic acetylcholine receptor reactive Th17 cells in patients with primary Sjögren's syndrome

Previously M3 muscarinic acetylcholine receptore (M3R) reactive Th1 cells were identified, and here, M3R reactive Th17 cells were identified in peripheral blood of primary Sjögren's syndrome patients using ELISpot method. To assess TCR repertoire overlap between identified M3R reactive Th17 cells, and salivary gland infiltrating T cells, we performed high throughput TCR sequencing of those cells from pSS patient. Overall design: From pSS patient who were detected positive for M3R reactive Th17 cells by ELISpot method, IL-17 producing CD4+ T cells were FACS sorted after stimulating with M3R peptide for 8 days. Salivary gland(SG) was taken at the time of diagnosis, and stored in RNA later solution. Then, we prepared libraries from these CD4+ T cell sample, together with salivary gland sample and performed unbiased high throughput TCR sequencing using an Ion Genestudio S5 next generation sequencer.