Fig. 8 | Nature Communications

Fig. 8

From: Hypoxia-inducible factor-1α is a critical transcription factor for IL-10-producing B cells in autoimmune disease

Fig. 8

HIF-1α expression in B cells regulates helper T cell polarization in vitro. a Naïve CD4 T cells isolated from WT mice were co-cultured with CD1dhiCD5+ B cells from WT mice with anti-IL-10 antibody or isotype control antibody at a 1:1 T/B ratio in a trans-well system for 3 days under T cell-polarizing conditions. Percentage of IFN-γ+CD4+(Th1) cells, IL-17+CD4+(Th17) cells, and IL-10+CD4+ (Tr1) cells were determined (n = 3 per group). b Naïve CD4 T cells isolated from WT mice were co-cultured with CD1dhiCD5+ B cells from WT mice (CD1dhiCD5+(WT)), Mb1creHif1af/f mice (CD1dhiCD5+Hif1a)), Mb1creHif1af/f mice after transduction of pDBR lentivirus (mock-CD1dhiCD5+Hif1a)), or Mb1creHif1af/f mice after transduction of pDBR-IL-10 lentivirus (IL-10-CD1dhiCD5+Hif1a)) at a 1:1 T/B ratio in a trans-well system for 3 days under T cell-polarizing conditions. Percentage of IFN-γ+CD4+(Th1) cells, IL-17+CD4+(Th17) cells, and IL-10+CD4+(Tr1) cells were determined (n = 3 per group). c Naïve CD4 T cells isolated from WT mice were co-cultured with CD1dhiCD5+ B cells from WT mice (CD1dhiCD5+(WT)) or Mb1creHif1af/f mice (CD1dhiCD5+Hif1a)) at a 1:1 T/B ratio in a trans-well system for 3 days. Percentage of IL-10+CD4+ T cells and Foxp3+CD4+ T cells were determined (n = 6 per group). Data are shown as mean ± s.e.m. Pictures are representative of three independent experiments. NS not significant; *P < 0.05, **P < 0.01, and ***P < 0.001(unpaired, two-tailed Student’s t-test)

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