Gene expression profiling of virus-specific CD8 T cells shows enrichment in IFN-I and CD28 driven genes.

(A) Experimental outline for comparing the transcriptional signature of virus-specific CD8 T cells. RNA-Seq was performed on FACS-sorted DbGP276+ CD8 T cells from spleen. (B) Cell purity following FACS-sorting of virus-specific CD8 T cells. (C) PCA comparing the transcriptional landscape of rescued CD8 T cells following PD-L1 blockade alone or combined LPS and PD-L1 blockade. (D) Heat map of the most differentially expressed genes between single and combined treatment. (E) GSEA plots demonstrating enrichment for IFN-I signaling genes in virus-specific CD8 T cells following combined therapy. (F) Radar plots showing expression of various IFN-I driven genes. (G) GSEA plots demonstrating enrichment for CD28 costimulation genes in virus-specific CD8 T cells following combined therapy. In panels E and G, DN and UP mean downregulated or upregulated, respectively, relative to previously identified transcriptional signatures used as reference. (H) Radar plots showing expression of various CD28 driven genes. The genes selected were shown to be enriched following CD28 costimulation in a prior publication [71]. Presented data are from one experiment, control (n = 3), PD-L1 therapy alone (n = 3), or combined LPS and PD-L1 therapy (n = 4) at day 15 post-treatment.