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 CD1d^hiCD5⁺ 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 CD1d^hiCD5⁺ B cells from WT mice (CD1d^hiCD5⁺(WT)), Mb1^creHif1a^f/f mice (CD1d^hiCD5⁺(ΔHif1a)), Mb1^creHif1a^f/f mice after transduction of pDBR lentivirus (mock-CD1d^hiCD5⁺(ΔHif1a)), or Mb1^creHif1a^f/f mice after transduction of pDBR-IL-10 lentivirus (IL-10-CD1d^hiCD5⁺(ΔHif1a)) 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 CD1d^hiCD5⁺ B cells from WT mice (CD1d^hiCD5⁺(WT)) or Mb1^creHif1a^f/f mice (CD1d^hiCD5⁺(ΔHif1a)) 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)