Journal: Nature Communications
Article Title: IL-6 receptor blockade corrects defects of XIAP-deficient regulatory T cells
Figure Lengend Snippet: Anti-IL-6R reduces the expression of IFN-γ in activated Xiap −/− Treg cells. a , b Anti-IL-6R decreases IFN-γ expression in re-stimulated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated with anti-CD3/CD28 and IL-2, or with additional IL-12 as indicated, in the absence or presence of anti-IL-6R (50 μg ml −1 ) for 4 days. iTreg cells were re-stimulated with TPA/A23187 for 24 h and IFN-γ production was determined by ELISA ( a ), or reactivated with TPA/A23187 for 5 h and the expressions of Foxp3 and IFN-γ were determined by intracellular staining ( b ). c , d Anti-IL-6R inhibits IFN-γ expression in human Treg cells. Control and human XIAP-knockdown iTreg cells were stimulated as in ( a , b ), with additional IL-6 as indicated, and secretion ( c ) or intracellular expression ( d ) of IFN-γ was determined. e Inability of anti-TNF or anti-IL-1R to inhibit the production of IFN-γ in activated Xiap −/− Treg cells. WT and Xiap −/− iTreg cells were stimulated, as described in ( a ), in the presence or absence of anti-TNF or anti-IL-1R (50 μg ml −1 each) for 4 days. Treg cells were re-stimulated with TPA/A23187 for 24 h and the secreted IFN-γ was determined by ELISA. f Anti-IL-6R rescues the impaired suppressive activity of Xiap −/− tTreg cells in vivo. CD45.2 + WT or Xiap −/− tTreg cells were co-transferred with CD45.1 + CD4 + CD25 - effector T cells into male CD45.1 + RagI −/− mice. Anti-IL-6R antibody (500 μg per mouse) was intraperitoneally administrated at day 0, followed by weekly dosing of 500 μg. The body weights of mice were monitored at the indicated time-points. *** P
Article Snippet: Recombinant mouse IL-2, human IL-2 and mouse IL-6 were purchased from eBioScience.
Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, In Vivo, Mouse Assay