Fig. 7: moDCs comprise three different subsets that are defined by characteristic protein expression profiles and differential susceptibilities to HCMV infection. | Nature Communications

Fig. 7: moDCs comprise three different subsets that are defined by characteristic protein expression profiles and differential susceptibilities to HCMV infection.

From: Human cytomegalovirus exploits STING signaling and counteracts IFN/ISG induction to facilitate infection of dendritic cells

Fig. 7

moDCs were differentiated from 16 independent donors, CLEC12A, CD1a, CD86, CCL18, CCL17, CCL22, CD115, CD88 and CD85d were immunolabeled and analyzed by flow cytometry. a UMAP of live cells showing heatmap coloring to indicate the abundance of each of the immunolabeling markers, data of one representative donor are shown. b Mock-treated moDCs were immunolabeled as described above and the three moDC subsets were discriminated by gating of CD1a−/CD86− (M1, blue dots), CD1a+ (M2, red dots) and CD86+ (M3, green dots) cells. c Frequencies and d relative fluorescence intensities (RFI) of each of the analyzed markers in the three subset gates were determined. Data represents mean ± SEM of 16 different donors from 7 independent experiments. Each dot represents a single donor. ****p < 0.0001, *p = 0.0182 using two-sided paired Wilcoxon signed-rank test (c) and ****p < 0.0001 (CD1a, CD86), ***p = 0.0010 (CCL17, CCL18), p = 0.0003 (CLEC12A), p = 0.0005 (CCL18, CCL22), **p = 0.0039 (CD115), p = 0.0093 (CCL18), 0.0020 (CD85d), p = 0.0015 (CCL17, CCL22), *p = 0.0137 (CD88) using two-sided paired Wilcoxon signed-rank test (d). e moDCs were infected with HCMV-NG, immunolabeled and analyzed as in a. UMAP of NG fluorescence and the 3 most discriminative moDC subset markers, i.e., CD1a, CD86, and CLEC12A. Insets show the expression of the above-mentioned markers only in the NG+ cluster. f Mock-treated moDC subsets were sorted using the gating strategy shown in (b) and analyzed morphologically (scale bar 50 µm). moDC cultures (g) or sorted moDCs (h) as described in b were infected with HCMV-NG and percentages of NG+ cells were determined 24 hpe. Data represents mean ± SEM of 6 different donors from 3 independent experiments. Each dot represents a single donor. *p = 0.0313 using two-sided paired Wilcoxon signed-rank test (g) and ± SEM of 3 different donors from 1 experiment. Each dot represents a single donor. i Quantification of STING protein expression from the subsets described in b. Data represents mean ± SEM of 7 different donors from 4 independent experiments. Each dot represents a single donor. *p = 0.0469 (M2 vs. M1), p = 0.0156 (M3 vs. M2), and p = 0.0313 (M3 vs. M1) using two-sided paired Wilcoxon signed-rank test.

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