Human miR-583 downregulates NK cell activation by silencing IL2Rγ during NK cell differentiation.

<p>(a) The expression of IL2Rγ was analyzed by FACS 14 days after miR-control or miR-583 transfection in differentiating NK cells and CD56⁺CD3⁻ gated NK cells. Gray, IgG control; blue, miR-control; red, miR-583. The absolute numbers of differentiated NK cells are shown in the parentheses (×10⁵). (b) The expression of p-STAT5 was analyzed by FACS 7 days after miR-control or miR-583 transfection in differentiating NK cells. Gray, IgG control; blue, miR-control; red, miR-583. (c) The expression levels of NK cell activation receptors in differentiating NK cells and CD56⁺CD3⁻ gated NK cells were analyzed by FACS 14 days after miR-control or miR-583 transfection. Gray, IgG control; blue, miR-control; red, miR-583. (d). Cytotoxicity of differentiating NK cells was determined by ⁵¹Cr release assay against K562 cells at the indicated effector/target (E:T) ratios. *, <i>p</i><0.1. The expression levels of CD107a in differentiating NK cells and CD56⁺CD3⁻ gated NK cells were analyzed by FACS. Gray, IgG control; blue, miR-control; red, miR-583. (e) Differentiating NK cells were stimulated with PMA/IO or IL-18. After 16 h, the supernatants were assayed for IFN-γ production by ELISA. *, <i>p</i><0.1. Values represent the mean % of positive cells ± S.E.M. of triplicates. The data are representative of three independent experiments performed using three different UCB samples and represent the mean values ± S.E.M. of duplicates.