Figure 2.

CX₃CR1 expression in DC after maturation. Mo-derived iDC were further cultured in presence of LPS (500 ng/ml)+1000 U/ml IFN-γ for 48 h to obtain mDC. Subsequently, the cells were stained with FITC-conjugated anti-CD1a mAb, PE-conjugated anti-CD83 mAb, PC5-conjugated anti-CD86 mAb and FITC-conjugated anti-HLA-DR mAb and then analyzed by flow cytometry. (a) Representative histograms for CD1a, CD83, CD86 and HLA-DR membrane expression in iDC or mDC are shown. Dotted histograms represent the isotype controls. (b) The percentages of positive cells for CD1a, CD83, CD86 and HLA-DR in iDC or mDC are shown. Each bar represents the mean±s.e.m. from 4–6 different donors. *P<0.05 and **P<0.01, by Mann–Whitney U test calculated for each marker individually. (c) CX₃CR1 expression in Mo, iDC or mDC. The results are expressed as percentage of positive cells. Each bar represents the mean±s.e.m. from 7–10 different donors. ***P<0.001, according to Kruskal–Wallis test (P<0.001) followed by the Mann–Whitney U test. (d) Mo were cultured up to 3 days in the presence of GM-CSF (50 ng/ml)+IL-4 (IL4DMo) or IL-13 (20 ng/ml) (IL13DMo) with or without IFN-γ (1000 U/ml). Subsequently, the cells were stained with FITC-conjugated anti-CX₃CR1 mAb and then analyzed by flow cytometry. The percentages of CX₃CR1 positive cells in every cell population are shown. Each bar represents the mean±s.e.m. from 2–4 different donors. DC, dendritic cell; GM-CSF, granulocyte-macrophage colony stimulating factor; HLA, human leukocyte antigen; IFN, interferon; LPS, lipopolysaccharide; mDC, mature dendritic cell; PE, phycoerythrin.