Figure 5. Cytokine production and TCR repertoire analysis of M3R Th17 cells.

(A) Representative FACS plot of 3 pSS ELISpot-positive patients (pSS1, -3, -5) shows that IL-17⁺CD4⁺ cells were increased together with IL-17⁺IFN-γ⁺ double-positive CD4⁺ cells after culturing PBMCs with M3R AA83-95 peptide for 8 days. (B) Statistical analysis of each subset in above 3 pSS. *P < 0.05, by Kruskal-Wallis test. (C) Sorting strategy of M3R AA83-95–induced IL-17⁺CD4⁺ T cells from pSS9. Dead cells are previously gated out. (D)TCR repertoire overlap analysis between LSG infiltrating T cells and peripheral M3R-reactive Th17 cells in 1 pSS ELISpot-positive patient (pSS9). Frequency of overlapping clones between samples are shown for the top 20 overlapping clones (colored), as well as the clones those frequency were below the top 20 (black) and nonoverlapping (gray) clones. Overlapping peripheral M3R Th17 cells TCR repertoire were seen in about 10% of LSG TCR repertoire. (E) Overlapping clones between each sample were plotted with log₁₀ frequencies in each sample. Each plot represents overlapping clonotype. Major clones in peripheral M3R Th17 cells were not as dominant in LSG, and vice versa.