Fig. 8: Effect of DAA treatment and HMTs/p53 modulation on HCV-specific CD8+ T cells from chronic patients.

a Glucose uptake (measured by the incorporation of the glucose analog 2-NBDG), percentage of depolarized mitochondria (by staining with the mitochondrial membrane potential sensitive dye JC-1), PD-1 expression, proteasomal function (by ProteoStat staining) and repressive H3K9me2 mark were assessed in virus-specific, dextramer-stained CD8+ T cells from T2/late HCV patients (n = 9) at baseline and at EOT, from T2/late resolved patients and from healthy controls after overnight anti-CD3/anti-CD28 stimulation (see Methods section for details). All data were analyzed with the Kolmogorov-Smirnov test, followed by Wilcoxon matched-pairs signed rank test (paired for chronic patients at baseline vs. EOT). Conversely, differences between multiple patient groups were evaluated with the non-parametric Kruskal-Wallis test; p-values were corrected for pairwise multiple comparisons, with the Dunn’s test. b PBMC from chronic HCV patients (T2/late; n = 6) at baseline and at EOT were stimulated for 40 h with HCV-NS3 peptides in the presence or absence of the EZH2 inhibitors GSK126 (GSK) and EPZ005687 (EPZ) (red dots), of the G9a inhibitors UNC0638 (UNC) and BIX01294 (BIX) (green dots), and of the p53 inhibitor pifithrin-alfa (p53) (blue dots), followed by co-staining for IFN-γ, IL2, and TNFα (left). c PBMC from baseline and EOT of T2/late chronic HCV patients (n = 11) were stimulated for 10 days with HCV-NS3 peptides as in b. Data in b and c are presented as the ratio (fold-change) between cytokine producing CD8+ T cells detected in treated vs. untreated cultures from individual patients; statistical analysis was performed with the Wilcoxon signed-rank test; horizontal lines represent median values.