Fig. 5: scRNAseq transcriptomic and TCR repertoire analysis of ex vivo MPXV-specific CD8⁺ T cells from vaccinated (n = 6) and convalescent (n = 4) individuals.

Tetramer⁺CD8⁺ T cells were used for single-cell transcriptomic and TCR repertoire analysis. A UMAP plot of tetramer⁺CD8⁺ T cells integrated based on patient of origin, identifying 4 clusters of cells. B Heatmap of the differentially expressed marker genes of the 4 cell clusters. C Beeswarm plot showing the log-fold distribution of cell abundance changes between convalescent and vaccinated samples. Neighbourhoods overlapping the same cell population are grouped together, neighbourhoods exhibiting differential abundance are coloured in red and are circled. D Violin plots of cluster 2 marker genes between vaccinated and convalescent samples. E and F Circos plots showing the TRAV-TRAJ and TRBV-TRBJ gene pairing for convalescent (E) and vaccinated (F) samples. G UMAP plot of the tetramer⁺CD8⁺ T cells, split by whether their TCR alpha (top) and beta (bottom) clonotypes are expanded. Differential expression between clusters was carried out using the MAST test. The Wilcoxon signed-rank test was used to test differences between convalescent and vaccinated samples and two-tailed P values were calculated. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.