Fig. 7

Macrophage differentiation and function is impaired in patients with WAS. Monocyte-derived macrophages form healthy donors (control) and patients with WAS were differentiated into M1 macrophages. a Expression of M1-specific markers was analysed by qPCR. b Naive CD4⁺CD25⁻ T cells were CFSE labelled and cultured in the presence of plate-bound anti-CD3 (2 μg/ml) and either control or WAS patient-derived M1 macrophages for 3 days. T-cell proliferation was determined by flow cytometer. Data are representative of two independent experiments with two patient samples. Monocyte derived macrophages form healthy donors (control) and patients with WAS were differentiated into M2r macrophages. c Expression of M2-specific markers was analysed by qPCR. d M2r differentiated monocyte-derived macrophages were restimulated with LPS for 4 h. Expression of pro-inflammatory genes were analysed by qPCR. e Naive CD4⁺CD25⁻ T cells were CFSE labelled and cultured in the presence of plate-bound anti-CD3 (2 μg/ml) and either control or WAS patient-derived M2r macrophages for 4 days. T-cell proliferation was determined by flow cytometer. Data are representative of two independent experiments with two patient samples. f Naive CD4⁺CD25⁻ T cells were cultured in presence of plate-bound anti-CD3 (2μg/ml), TGF-β (2 ng/ml) and either control or WAS patient-derived M2r macrophages for 5 days. Treg generation was examined by analysing FOXP3 expression using flow cytometry. Data are representative of two independent experiments with two patient samples. g Naive CD4⁺CD25⁻ T cells from independent donor were cultured in the presence of plate-bound anti-CD3 (2 μg/ml) and either control or WAS patient-derived M2r macrophages for 3 days. Cells were restimulated with PMA (10 ng/ml), ionomycin (500 ng/ ml) in the presence of GogiStop for last 4 h. Intracellular cytokine expression was analysed by flow cytometry. Data are representative of two independent experiments with two patient samples