Fig. 1: Single-cell RNA sequencing reveals heterogeneity of human monocyte-derived dendritic cells.

Blood-derived CD14⁺ monocytes were differentiated to moDCs and exposed to HCMV-NG (NeonGreen) at MOI 6. a Flow cytometry analysis of mock-treated and HCMV-NG exposed moDCs 8 and 24 hours post virus exposure (hpe). b Schematic depiction of the experimental setup (symbols from BioRender). Mock-treated and HCMV-NG exposed moDCs were labeled 8 hpe with anti-CD45-ADT and anti-HLA-DR-ADT antibodies, respectively, and pooled prior to scRNA-seq. This experiment was performed in four runs with moDCs from two donors and with two independent virus preparations. c, d Data from all four scRNA-seq runs were combined for non-linear dimensionality reduction (UMAP) and unsupervised clustering (bordered and numbered areas). Log normalized feature counts are shown for CD45-ADT (cells shown in blue) (c) and HLA-DR-ADT (cells shown in orange) (d). Two clusters (shown without borders) were identified to be composed of doublets and were removed for further analysis. e UMAP visualization of SCTransform normalized feature counts for the viral UL123/NeonGreen gene (cells shown in green). f Donor #1 and g donor #2 comprised 3 mock treated clusters (M1-3, CD45-ADT⁺), 4-5 bystander clusters (B1-4, HLA-DR-ADT⁺/UL123^low), and 1 productively infected cluster (P, HLA-DR-ADT⁺/UL123^high). Different colors show clusters with divergent gene expression profiles between mock, bystander, and productively infected moDCs from the two different analyzed donors.