Figure 3

Expansion of Th17 cells in PBMCs of patients with active SSc. (A) Human PBMCs were labeled with antibody against lymphocytes (anti-CD3 and -CD8). IL-17-expressing cells were detected by intracellular cytokine staining assay, and the percentage of IL-17⁺ cells among CD3⁺CD8^- T cells was determined with flow cytometry. (B) Results of flow-cytometric analysis of Th17 cells in patients with active SSc (n = 13), patients with stable SSc (n = 32), and controls (n = 24). (C) Longitudinal monitoring of Th17 cells in 10 patients. The percentage of Th17 cells was measured initially during active SSc and again after resolution after treatment. (D) Real-time RT-PCR analysis of IL-17, Foxp3, and RORγt mRNA expression in freshly isolated PBMCs of patients with active SSc (n = 13), patients with stable SSc (n = 32), and controls (n = 24). (E) Positive correlation between the proportion of Th17 cells and clinical severity in active SSc patients, scored by using the Valentini score (n = 13). r = 0.675, P < 0.01. (F) Results of flow cytometric analysis of CD4⁺CD25⁺CD127^- Treg cells in patients with active SSc (n = 13), patients with stable SSc (n = 32), and controls (n = 24).