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. 2022 Apr 12;13:1970. doi: 10.1038/s41467-022-29539-3

Fig. 3. Regulatory T cells (Tregs) are enriched and have enhanced suppressive functions in synovial fluid (SF) of patients with arthritis as an immune-related adverse event.

Fig. 3

a, b Identification of two subclusters of Tregs and compartmental distribution of Treg subclusters. c Quantification of the subclusters and comparison of their cellular fractions between 8 peripheral blood (PB) and SF samples. Two-sided unpaired t test. Subcluester 1, ****P < 0.0001; Subcluster 2, ****P < 0.0001. Bars indicate the mean and SEM. d Volcano plot showing differentially expressed genes between PB Tregs and SF Tregs. FC, fold change. See Supplementary Data 4 for all differentially expressed genes (DEGs) and Supplementary Data 5 for DEGs with log2 fold changes >0.5 or < −0.5 and -log10 P > 15. e Representative flow cytometry plots of Tregs (left panels) and quantification analysis (right panel). n = 4 PB and matching SF samples. Two-sided paired t test. *P = 0.0354. f Mean fluorescence intensity (MFI) of CD25, FOXP3, CTLA-4, and PD-1 on non-Tregs (CD25lo CD127hi/lo) and Tregs (CD25hi CD127lo). Representative flow cytometry plots (upper panels) and quantification analysis (lower panel) are shown. n = 4 PB and matching SF samples. Two-sided paired t test. CD25, **P = 0.0030; FOXP3, *P = 0.0224; CTLA-4, *P = 0.0460; PD-1, *P = 0.0116. g Suppression of polyclonal response of naïve CD4+ T cells with autologous PB Tregs or SF Tregs. The percentage of suppression of cell proliferation (CFSE dilution) and cytokine production were calculated as described in the Methods. Parallel experiments were performed with PB Tregs from patients in the no-irAE group. n = 4 PB and matching SF samples from arthritis-irAE patients; n = 5 PB samples from no-irAE patients. One-way analysis of variance. Cell proliferation, ***P = 0.0002, ****P < 0.0001; IFNγ production, *P = 0.0201, ***P = 0.0003; IL-2 production, *P = 0.0130, **P = 0.0011. Bars indicate the mean and SEM. IFNγ, interferon gamma; IL-2, interleukin-2. Source data are provided as a Source Data file.