Fig. 5
IL-27 p28 deficiency restrains Treg cell differentiation via the IFN-γ/STAT1 signaling pathway. a, b Splenocytes from WT or CD11c-p28f/f mice were induced for Treg polarization in the presence or absence of IL-27 p28 antibody (10 μg/ml). Representative figures and summary data of the frequency of Tregs are depicted. c BALB/c recipients were injected with 1 × 107 TCD-BM cells from CD45.1 mice, along with an equal number of T cells (2 × 106) from CD11c-p28f/f mice and CD45.1 congenic mice. Treg cell reconstitution was evaluated in vivo 14 days post transfer. d BALB/c recipients were transplanted with 5 × 106 TCD-BM cells plus 1 × 106 conventional T cells (Tconv) either from WT or CD11c-p28f/f mice and transferred with 7.5 × 105 Treg cells from WT or CD11c-p28f/f mice respectively. The overall survival curve is depicted (n = 10 per group). e BALB/c recipients were transplanted with 5 × 106 TCD-BM cells plus 1 × 106 Tconv cells from WT donors and transferred with or without 7.5 × 105 Tregs from WT or CD11c-p28f/f mice (n = 10 per group). f–h Anti-IFN-γ, anti-IL-6R, or anti-IL-12 were added during Treg polarization at a concentration of 10 μg/ml. Percentage of Treg cells was detected by FACS (n = 3 per group). i IFN-γ production in the supernatant of polarized Tregs was detected by ELISA (n = 4 per group). j The expression of phosphorylated STAT1 among Treg populations is shown (n = 5 per group). k STAT1 inhibitor (10 μM) was added during Treg polarization. Percentages of Treg cells were detected by FACS (n = 3 per group). l, m BALB/c recipients were transplanted with 1 × 107 WT BMs together with 5 × 106 splenocytes from either WT or CD11c-p28f/f mice. Recipients were injected with αIFN-γ (250 μg per mouse) after BMT. Percentages of Tregs were detected 2 weeks after BMT (n = 4 per group). The overall survival curve is depicted (n = 10 per group). Data are representative of three independent experiments and presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001
