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. 2016 Oct 14;7:423. doi: 10.3389/fimmu.2016.00423

Figure 1.

Figure 1

IL-4 downregulates Mincle mRNA expression in human antigen-presenting cells. (A) Monocyte-derived DC express less MINCLE than monocyte-derived macrophages. Human CD14+ cells were differentiated for 6 days in the presence of M-CSF or GM-CSF (to obtain macrophages) or GM-CSF + IL-4 (to obtain dendritic cells). After that, MINCLE mRNA levels were measured using qPCR. The data are depicted as mean ± SD of ΔCT values (Ct[PPIA] − Ct[MINCLE]) of cells from eight independent blood donors. (B) Reduced expression in DC is restricted to MINCLE, MCL, DECTIN2, and CLEC12A. Gene expression of human GM-CSF-elicited macrophages was compared to dendritic cells (induced via GM-CSF + IL-4) after 6 days of differentiation using qPCR. The data are depicted as mean ± SD of ΔΔCT values (using GM-CSF Mϕ as calibrators) from 6 independent blood donors. (C) IL-4 is sufficient to reduce MINCLE expression in human CD14+ cells. CD14+ peripheral blood mononuclear cells were stimulated as indicated. MINCLE expression was determined using qPCR. The data are depicted as mean ± SD (n = 2 for d0, n = 4 for d0.25, and n = 6 for all other time points). Asterisks indicate significant p-values for the comparison “IL-4 vs. untreated”, crosses for the comparison “GM-CSF + IL-4 vs. GM-CSF”. (D) Dose-dependence of IL-4 effect. CD14+ peripheral blood mononuclear cells were stimulated with increasing concentrations of IL-4 for 24 h. Mean ± SD of ΔCT values for MINCLE expression of cells from four independent donors. (E) Human CD14+ monocyte-derived M-CSF or GM-CSF elicited macrophages and dendritic cells were stimulated with IL-4 or left untreated for 48 h. Subsequently, MINCLE expression was determined using qPCR. The data are depicted as mean ± SD of cells from four independent blood donors. Statistical significance was tested using a paired t-test. *p < 0.05, **p < 0.01, ***p < 0.001, †††p < 0.001.