CD8 T cells expressing DN Ikaros have enhanced expression of CD25 (IL-2 receptor alpha chain) in an IL-2 independent manner.

<p>(A) Relative abundance of CD25 in CD8 T cells determined by flow cytometry, analyzing cells that were mock transduced (gray), transduced with control vector (blue, Thy1.1+ cells in MiT transduced cultures) or with DN Ikaros retrovirus (red, Thy1.1+ cells in MiT-DN Ikaros transduced cultures). Cells were cultured in mIL-2 (20 ng/mL) (top), mIL-12 (5 ng/mL) (middle) or no exogenous cytokine (bottom). (B) DN Ikaros expression does not enhance CD69 expression in activated CD8 T cells cultured in mIL-12 (5 ng/mL). Data representative of two (IL-2) to five (IL-12) independent experiments. (C) Anti-IL2 antibody effectively neutralizes IL-2 in vitro, as measured by decreased cell size (forward scatter, FSC) or decreased CD25 following culture of activated CD8 T cells in IL-2 (20 ng/mL), comparing treatment with either an isotype control antibody (black) or an anti-IL2 antibody (gray). (D) DN Ikaros-transduced cells cultured in IL-12 (5 ng/mL) have enhanced CD25 expression in an IL-2 independent manner. Data show CD25 expression in DN Ikaros-transduced cultures at day 4 or 6 of culture, in cells treated with isotype control (black) or anti-IL2 antibody (red), relative to cultures treated with an isotype antibody (gray); results representative of two independent experiments.