Research Article

IL-27 Driven Upregulation of Surface HLA-E Expression on Monocytes Inhibits IFN- Release by Autologous NK Cells

Figure 2

Modulation of HLA-G and HLA-E expression in human monocytes and functional assays on NK cells. (a) HLA-G and -E expression was investigated in monocytes cultured with medium alone (white bars), 100 ng/mL IL-27 (light grey bars), or 100 ng/mL IL-30 (grey bars). Results are represented as MRFI. Histograms represent mean of six different experiments ± SD. Asterisks indicate statistical significance. One representative FACS analysis is reported in (b). Grey profiles show staining with irrelevant fluorochrome and isotype matched mAb. Black profiles show staining with specific mAb. (c) IFN- was analyzed in supernatants from NK cells cultured with (i) medium alone (stripped bar), (ii) in the presence of coated anti-NKp46 mAb (black bar) or in the presence of coated anti-NKp46 mAb and autologous monocytes, (iii) untreated (white bar), (iv) pretreated with IL-27 (light grey bar), or (v) pretreated with IL-30 (grey bar). Results are represented as ng/mL IFN- . Histograms represent mean of six different experiments ± SD. Asterisks indicate statistical significance. (d) Expression of CD107a was evaluated on NK cells cultured with (i) medium alone, (ii) target cells (K562) at 4 : 1 effector : target ratio, and target cells (K562) in the presence of autologous monocytes, (iii) untreated, (iv) pretreated with IL-27, or (v) pretreated with IL-30. One representative experiment out of three performed is shown. The percentage of CD107a+ cells (gating on NK cells using physical parameters) is indicated.
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