Fig. 3: Analysis of circulating CD3⁻ immune cell populations in healthy donors (HD), patients with newly diagnosed (ND) and relapsed/refractory (R/R) cHL and detailed analysis of NK cell subsets.

a Annotated UMAP of CD3⁻ single-cell expression profiles (n = 229,670 cells) by cell type. Each cluster is labeled with a distinct color and unique number, beginning with the most abundant cluster (Cluster 0). b Feature plots of selected cell lineage and differentiation markers. c Dot plot with relative expression of selected genes in NK cell clusters. Displayed genes were curated from the top differentially expressed genes that defined clusters using a two-sided Wilcoxon rank sum test, adjusted for multiple comparisons (adj p < 0.05, fold change > 1.75) and supplemented with relevant markers based on a priori knowledge. The size of the dot indicates the percentage of marker-expressing cells in each cluster and the z-score reflects mean marker expression across the clusters. d Relative abundance of NK cell clusters in HD (n = 13) and patients with ND cHL (n = 11). Differences between HD and patients with ND cHL were assessed by a two-sided Wilcoxon rank sum test. P values that remained significant after Benjamini–Hochberg correction FDR < 0.1 are noted (*). e Lineage trajectory analysis of NK cell populations (n = 40,844 cells) by STREAM overlaid on UMAP (left panel); Subway map plot (middle panel) showing all individual NK cells; Stream plot (right panel) visualization of cell density along different trajectories. At a given pseudotime, the width of each branch is proportional to the total number of cells. f Stream plots visualization of the selected marker genes, representing stages of NK cell maturation and/ or function. Source data are provided as a Source Data file.