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N. van Leeuwen-Kerkhoff et al.
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Figure 5. Functional capacities of dendritic cell subsets and slan+ monocytes. (A) Maturation capacity of cDC2 and slan+ monocytes in myelodysplastic syndrome (MDS) bone marrow (BM) upon toll-like receptor (TLR)-stimulation. Expression levels of CD80, CD86 and HLA-DR were assessed by flow cytometry at baseline (T=0) and after overnight stimulation with LPS + R848 (+). Mean fluorescence intensity (MFI) values of these three markers were measured. Median values of 4-7 exper- iments are shown. (B) Up-regulation of CD80, CD86 and HLA-DR after overnight TLR-stimulation in normal bone marrow (NBM)- (n=4) and MDS-derived (n=4-7) cDC2 and slan+ monocytes. Median values are shown. (C) Cytokine secretion assay. Culture supernatants of healthy (n=4) and MDS (n=10) BM-derived unstimulated and stimulated cDC2 and slan+ monocytes were analyzed for the presence of different cytokines by cytometric bead array. Median values are shown. (D) Allogeneic mixed leukocyte reaction (MLR). Peripheral blood lymphocytes (PBL) were labeled with carboxyfluorescein succinimidyl ester (CFSE) and co-cultured with healthy (n=2-4) or MDS-derived (n=1-5) stimulated cDC1, cDC2 or slan+ monocytes. The percentage of CFSE-diluted T cells was determined by flow cytometry. Median values of dif- ferent experiments are shown. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. pg: picogram.
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haematologica | 2022; 107(3)