Figure 7
IL-4 inhibits TGF-β-induced adaptive Tr-cell conversion. MLN-DCs were loaded with 0.2 μg ml−1 OVA peptide during 6 h. After washing, 2.0 × 104 MLN-DCs were incubated with 5 × 105 CFSE-labeled CD4+KJ1.26+ T cells in the presence of TGF-β and retinoic acid during 96 h and analyzed for Foxp3 expression and CFSE content by flow cytometry. (a) Representative flow cytometric analysis of Foxp3 expression in the presence or absence of exogenous recombinant IL-4 (b) combined data n=3. (c) To determine whether the IL-4 that is produced during COX-2 inhibition directly interferes with Foxp3 expression, cells were cultured with NS-398 and anti-IL-4 (10 μg ml−1; purified from 11B11 hybridoma). At 96 h, the cells were analyzed for Foxp3 expression. CFSE, 5,6-carboxy-succinimidyl-fluoresceine ester; COX-2, cyclooxygenase-2; DCs, dendritic cells; IL, interleukin; MLN, mesenteric lymph node; OVA, ovalbumin; RA, retinoic acid; TGF-β, transforming growth factor-β.*P<0.05
