Fig. 3

CD8⁺ T cells from LN patients displayed a greater tendency to differentiate into CD8⁺CD103⁺ cells. CD8⁺ T cells isolated from the PBMCs of patients with LN or HCs were triggered by anti-CD3/CD28 Dynabeads and 10 ng/ml TGF-β for 3 days to induce CD103⁺ cell differentiation. A and B: CD103 expression on CD8⁺ T cells was assessed by flow cytometry. Representative contour plots and percentages of CD8⁺CD103⁺ T cells are displayed (n = 5). C and D: Activated caspase-1 levels were quantified by flow cytometry, and representative images concentrating on CD8⁺CD103⁺ T cells are presented (n = 5). E–I: Following 3 days of stimulation, CD8⁺CD103⁺ cells from patients with LN or HCs were sorted. E–H: Cells were lysed and analyzed for NLRP3, pro caspase-1, caspase-1 p20, full length GSDMD, cleaved GSDMD, and GAPDH by western blotting. Representative bands and their densities are presented (n = 3). I: Sorted CD8⁺CD103⁺ cells were further cultured for an additional three days with anti-CD3/CD28 Dynabeads in the absence of TGF-β. The culture supernatant was then harvested, and IL-1β levels were assessed by ELISA (n = 3). Data are displayed as the mean ± SEM. *p < 0.05, **p < 0.01, and ****p < 0.0001, unpaired Student’s t-test