Figure 5: DC expression of Mbd2 is vital for optimal Th2 induction and development.

(a,b) CD4⁺GFP⁻ T cells from KN2xIL-10eGFP or KN2xIL-13eGFP mice were cultured for 3 days with WT (blue) or Mbd2^−/− (red) BMDCs, anti-CD3 mAb±IL-4 (Th2 conditions) and assessed for IL-4 (huCD2) protein production, and IL-10 and IL-13 mRNA expression by flow cytometry (five to six replicate culture wells, one of six experiments). (c) CD4⁺GFP⁻ T cells from KN2xIL-13eGFP mice were cultured for 3 days with WT (grey) or Mbd2^−/− (black) BMDCs, anti-CD3 mAb±IL-4 (Th2 conditions) and supernatants assessed for IL-10 and IL-13 protein secretion by ELISA. (d,e) CD4⁺GFP⁻ T cells from IL-10eGFP mice were cultured for 3 days with WT (grey) or Mbd2^−/− (black) DCs, anti-CD3 mAb±IL-12 (Th1 conditions) or IL-6/TGF-β/IL-23 (Th17 conditions) and supernatants assessed for IFNγ and IL-17 protein secretion by ELISA or IL-10 mRNA expression was determined by flow cytometry (four replicate wells per group, one of three experiments). (f,g) WT (grey) or Mbd2^−/− (black) BMDCs were cultured overnight in medium alone (M) or with SEA and injected s.c. into WT mice. Seven days later the pLNs were harvested, cells restimulated for 72 h with SEA and cytokine secretion assessed by ELISA (four to five mice per group, one of three experiments). Bar graphs show mean+s.e.m. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 (ANOVA). mAb, monoclonal antibody; s.c., subcutaneously; TGF-β, transforming growth factor-β.