Fig. 2
WJ-Muse display immunosuppressive properties. A Left: Representative flow analysis of HLA-G5 (soluble human leukocyte antigen-G5 marker), HLA-G1 (membrane-bound human leukocyte antigen-G1 marker), HLA-DR (MHC Class II, human leukocyte antigen marker) and PD-L1 (human Programmed death-ligand 1) in naive WJ-Muse. IgG were used as reference controls. Right: Representative flow analysis of HLA-G1, HLA-DR and PD-L1 in TNFα/IFNγ-primed WJ-Muse compared to naive WJ-Muse. B Evaluation of IDO, Cox2, PD-L1 and TGFβ1 expression level by quantitative real time PCR, in TNFα/IFNγ-primed WJ-Muse compared to naive WJ-Muse. Data are represented with means ± SEM, **p < 0.01; ***p < 0.001; ****p < 0.0001 (unpaired Student’s t test). C 105 human peripheral blood mononuclear cells (hPBMC) or murine spleen lymphocytes (mSL) were co-cultured with 104 WJ-Muse in order to evaluate their allogeneic or xenogeneic immune privilege. Frequency and proliferation of BrdU-labeled CD3+ T-cells were analyzed by flow cytometry and compared to hPBMC or mSL alone. Data are represented with means ± SEM. D Human peripheral blood mononuclear cells (hPBMC) or murine spleen lymphocytes (mSL) activated with concanavalin A were co-cultured with 5 × 103, 10 × 103 or 20 × 103 WJ-Muse to determine their allogeneic and xenogeneic immunosuppressive potential, respectively. Frequency (top) and proliferation (bottom) of human (left) or murine (right) BrdU-labeled CD3+ T-cells were analyzed by flow cytometry and compared, respectively, with activated hPBMC or mSL alone. Data are represented with means ± SEM, *p < 0.05, **p < 0.01 (two-tailed Mann–Whitney U test). E Arginase-1 (Arg1) and Nitric oxide synthase-2 (Nos2) mRNA expression were measured in bone marrow-derived macrophages (BMDM), which have been co-cultured for 7 days with WJ-Muse. Data are represented with mean ± SEM