Identification of CD80, CD86, and PD-L1 as potential therapeutic targets in cHL. (A) Schematic overview of components of the used multimodal target screening approach in cHL. (B) Volcano plot illustrating the log2 average fold change and P values of surface antigens absent on T cells and differentially overexpressed on HRSCs compared with GCB. Microarray data (GSE12453) were obtained from the gene expression omnibus¹⁷ (cHL, n = 12; RLN, n = 5). Labeled genes in red passed all filters (surface expression, absent on T cells), whereas non-highlighted gray dots indicate overexpressed genes that did not pass the filter thresholds. (C) Heat map visualizing the expression of the identified target antigens on microdissected control cells (left) or HRSCs (right). (D) Comparison of absolute densities of indicated antigens measured using flow cytometry on a panel of cHL cell lines (L-428, L-540, KM-H2) with that of the control cell line Nalm-6. Plotted is the pooled fold change ± standard error of the mean (SEM) of absolute molecule count in comparison to an isotype control stain of 3 different cHL cell lines. Statistical significance was calculated using 2-way analysis of variance (ANOVA) with Sidak multiple comparison correction. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. ns, P > .05. (E) Single-cell cross-organ off-target transcriptomic atlas screening for CD80, CD86, PD-L1, and CD30. The transcriptomic atlas consists of a total of 2.5 million sequenced cells comprising 11 different organs. A detailed summary of all used data sets is provided in the supplementary Material.