Fig. 2: Comparative analyses of CD3⁺CD8^- (CD4⁺ enriched) clusters in healthy donors (HD), patients with newly diagnosed (ND) and relapsed/ refractory (R/R) cHL.

a TCR repertoire diversity, as determined by Chao1 diversity index, in CD4⁺ naïve/CM T cell clusters from HD (n = 13) and patients with R/R cHL at baseline [C1D1] stratified by the best overall response (BOR) to PD-1 blockade. CR Complete response, PR partial response, PD progressive disease. Number of patients with Chao1 diversity data per cluster: Cluster 0, HD (n = 13), CR (n = 8), PR (n = 5), PD (n = 6); Cluster 1, HD (n = 13), CR (n = 9), PR (n = 5), PD (n = 6); Cluster 11, HD (n = 13), CR (n = 8), PR (n = 5), PD (n = 6); Cluster 12, HD (n = 13), CR (n = 8), PR (n = 4), PD (n = 6); Cluster 15, HD (n = 13), CR (n = 8), PR (n = 4), PD (n = 6); Cluster 18, HD (n = 13), CR (n = 9), PR (n = 4), PD (n = 4); Cluster 21, HD (n = 13), CR (n = 7), PR (n = 2), PD (n = 3). b Relative abundance of CD4⁺ naïve/CM T cell clusters in HD (n = 13) and patients with R/R cHL on treatment [C4D1] (n = 20) stratified by BOR to PD-1 blockade, CR (n = 9), PR (n = 5), PD (n = 6). c Dot plot with relative expression of selected genes associated with T cell naïveté and activation in all CD4⁺ naïve/CM clusters (Cluster 0, 1, 9, 11, 12, 15, 18, 21, 24) in HD (n = 13), patients with ND cHL (n = 11) and patients with R/R cHL at baseline [C1D1] (n = 20) and C4D1 (n = 20), stratified by BOR. The size of the dot indicates the percentage of marker-expressing cells and the z-score reflects mean marker expression across cohorts. d Relative abundance of cHL-specific T cell populations. e Venn diagram illustrating the shared clonotypes between CD4⁺ CTLs (Cluster 5, n = 992 clonotypes) and IFN-responsive CD4⁺ CTLs (Cluster 22, n = 231 clonotypes) in patients with cHL. f, g Relative abundance of gamma delta VD2 T cells (Cluster 23) and cycling CTLA4⁺ T cells (Cluster 19). d, f, g HD (n = 13), ND (n = 11), C1D1 and C4D1: CR (n = 9), PR (n = 5), PD (n = 6). h Dot plot illustrating the mean expression (color) and percentage of cells (dot size) expressing genes that positively identified cycling CTLA4⁺ T cells (Cluster 19) in comparison to the additional CD3⁺CD8⁻ clusters. i Cell segmentation and phenotype map for a representative multiplex IF image of ND cHL (magenta, HRS cells; yellow, CD4⁺CTLA4⁺Ki67⁺ cells; blue outline, FOXP3⁺ cells; gray-fill, Other cells less than 75 μm from HRS cells; gray-outline, Other cells greater than or equal to 75 μm from HRS cells). The experiment was performed in 9 ND cHL samples. j Relative abundance of CD4⁺CTLA4⁺Ki67⁺ and CD4⁺CTLA4⁺Ki67⁺FOXP3⁻ cells in relation to HRS cells (red dots, less than 75 μm from HRS cells; blue dots, greater than or equal to 75 μm from the HRS cells) in patients with ND cHL (n = 9). The indicated nominal p values were calculated by using a two-sided paired T-test and adjusted for multiple comparisons using the Benjamini-Hochberg method; p values that remained significant are noted (*). a, b, d, f, g Differences between HDs and patients with ND cHL were assessed by a two-sided Wilcoxon rank-sum test. The one-sided Cuzick trend test was used to compare patients with R/R cHL by BOR (CRs, PRs and PDs). Nominal p values that were significant (p < 0.05) are listed and those that remained significant after Benjamini–Hochberg correction FDR < 0.1 are noted (*). a, b, d, f, g, j All box plots generated in R display the 25th and 75th percentiles (lower and upper hinges), median values, and whiskers. The whiskers extend from the hinges to the largest/smallest values within 1.5 times the interquartile range (IQR) from the hinge. Data points beyond the end of the whiskers are plotted individually. Source data are provided as a Source Data file.