Figure 6.

IL-17C regulates both iTreg and nTreg cells during aGVHD. (A) Plate was coated with anti-CD3 and anti-CD28 overnight at 4°C. Naïve CD4⁺ T cells (CD4⁺CD62L⁺CD44⁻GFP⁻) were sorted from FoxP3-eGFP mice and seeded into 96-well plate in the presence or absence with rmIL-17C. Generation of iTreg cells were induced by rhIL-2 combined with rmTGF-β and examined 5 days later by flow cytometry. (B) BALB/C recipients were injected with IL-17C or control plasmid. 3 days later, recipients were lethally irradiated and transplanted with 1 × 10^∧7 bone marrow cells from CD45.1 mice and 2 × 10^∧6 naïve CD4⁺ T cells (CD45.2⁺CD4⁺CD62L⁺CD44⁻GFP⁻) from FoxP3-eGFP mice. Generation of iTregs were determined in H2-Kb⁺CD45.2⁺CD45.1⁻CD4⁺ populations at 10 days post transplantation in spleen; n = 5 per group. (C) BALB/C recipients were hydrodynamically injected with IL-17C plasmid or vector control. 3 days later, recipients were lethally irradiated and transplanted with 1 × 10^∧7 bone marrow cells together with 3 × 10^∧6 splenocytes from CD45.1 mice and 5 × 10^∧5 nTreg cells (CD45.2⁺CD4⁺GFP⁺) from FoxP3-eGFP mice. GFP expression was determined in H2-Kb⁺CD45.2⁺CD45.1⁻CD4⁺ populations at 10 days post transplantation in spleen; n = 3 per group. Data are representative of two experiments and presented as mean ± SEM. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001.