Fig. 9

Aggravating EAE in Hif1a-deficient mice is rescued by ecotopic expression of IL-10 in Hif1a-deficient CD1d^hiCD5⁺ B cells. a Clinical score of Mb1^creHif1a^f/f mice immunized with MOG_35-55 after adoptive transfer of CD1d^hiCD5⁺ B cells from WT mice (CD1d^hiCD5⁺(WT)) (n = 5), Mb1^creHif1a^f/f mice (CD1d^hiCD5⁺(ΔHif1a)) (n = 6), Mb1^creHif1a^f/f mice after transduction of pDBR lentivirus (mock-CD1d^hiCD5⁺(ΔHif1a)) (n = 5), or Mb1^creHif1a^f/f mice after transduction of pDBR-IL-10 lentivirus (IL-10-CD1d^hiCD5⁺(ΔHif1a)) (n = 5). b Pathophysiological pictures and quantifications of inflammatory loci (arrows) and demyelinated area (dashed line) in the spinal cord sections from Mb1^creHif1a^f/f mice immunized with MOG_35-55 after adoptive transfer of CD1d^hiCD5⁺(WT) (n = 5), CD1d^hiCD5⁺(ΔHif1a) (n = 6), mock-CD1d^hiCD5⁺(ΔHif1a) (n = 5), or IL-10-CD1d^hiCD5⁺(ΔHif1a) cells (n = 5). Scale bars, 500 μm. c IL-17, IFN-γ, IL-10, and TGF-β expression by splenocytes isolated from Mb1^creHif1a^f/f mice immunized with MOG_35-55 after adoptive transfer of CD1d^hiCD5⁺(WT) (n = 5), CD1d^hiCD5⁺(ΔHif1a) (n = 6), mock-CD1d^hiCD5⁺(ΔHif1a) (n = 5), or IL-10-CD1d^hiCD5⁺(ΔHif1a) cells (n = 5) followed by in vitro re-stimulation with MOG_35-55 (10 μM) for 48 h. d Representative plots and quantification of IL-17⁺CD4⁺(Th17), IFN-γ⁺CD4⁺(Th1), IL-10⁺CD4⁺(Tr1), and CD25⁺Foxp3⁺CD4⁺(Treg) cells in draining lymph nodes from Mb1^creHif1a^f/f mice immunized with MOG_35-55 after adoptive transfer of CD1d^hiCD5⁺(WT) (n = 5), CD1d^hiCD5⁺(ΔHif1a) (n = 6), mock-CD1d^hiCD5⁺(ΔHif1a) (n = 5), or IL-10-CD1d^hiCD5⁺(ΔHif1a) cells (n = 5). 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 (two-way analysis of variance with Bonferroni’s post-test (a) or two-tailed unpaired Student’s t-test (b–d)) (see also Supplementary Figure 8)