Fig. 4: IFNB1 correlates with viral IE gene expression, whereas the progression of viral infection inhibits IFNB1, but not IFNL1 induction.

a moDCs were exposed to HCMV-NG and supernatants were harvested completely and replenished with fresh medium 8, 16 and 24 hpe to determine the IFN-α, IFN-β or IFN-λ content by ELISA methods. Mean ± SEM of 6 different donors from 2 independent experiments. UMAP embeddings showing expression levels (SCTransform normalized values) of IFNB1 (b) and IFNL1 (c). d Spearman´s correlation coefficients of each individual viral gene with IFNB1 (x-axis) and IFNL1 (y-axis) expression for clusters B1-3 (1st graph), B4 (2nd graph), and P (3rd graph). White areas indicate statistically significant regions (p < 0.01, approximate two-sided t-test, Benjamini-Hochberg multiple testing correction). moDCs were treated one day before HCMV-NG exposure (−1) or at the time of HCMV-NG exposure (0) with e IFN-α2b (lavender bars), IFN-β (pink bars), or IFN-λ1 (blue bars), and f ADU-S100 (ADU) (green bars), or tumor necrosis factor (TNF) (purple bars), and NG⁺ cells were quantified by flow cytometry one (1) day post HCMV-NG exposure (dpe). Values for NG⁺ cells after HCMV-NG exposure and the treatment as indicated are shown relative to values for NG⁺ cells after HCMV-NG exposure without any other treatment, which were set to 100%. Values for HCMV-NG exposed cells without any other treatment are the same in e and f as the experiments were performed simultaneously. e Mean ± SEM of 7 different donors from 3 independent experiments. Each dot represents a single donor. *p = 0.0156 (−1 IFN-α vs. untreated), p = 0.0313 (−1 IFN-β vs. untreated) using two-sided paired Wilcoxon signed-rank test. (f) Mean ± SEM of 13 different donors from 6 independent experiments. *p = 0.0266, **p = 0.0078 using two-sided paired Wilcoxon signed-rank test. g M1, M2 and M3 moDC subsets were FACS sorted and left untreated (grey bar) or treated with ADU-S100 (ADU) or tumor necrosis factor (TNF) (green and purple bar, respectively) followed by exposure to HCMV-NG for 4 h. The genomic DNA was extracted from the soluble nuclear fraction and the relative abundance of HCMV genomes in relation to the housekeeping gene GAPDH was analyzed by qPCR. Mean ± SEM of 4 different donors from 1 experiment.