Fig. 1.

γ-H2AX (Ser-139) expression on circulating CD8+ T lymphocytes in healthy controls and HCV-infected subjects. (A) Gating strategy for γ-H2AX; cells within the live lymphocyte gate by scatter characteristics (left panel) and positive staining for CD8 (centre panel) were studied. Isotype control staining pattern in filled histogram; positive control staining pattern from irradiated cells in dashed histogram, and experimental sample in bold histogram (right panel). (B) Association between telomere length in CD8 + CD45RO+ subsets and γ-H2AX expression. Correlation by Spearman’s rank. (C) γ-H2AX levels in CD8+ T lymphocytes (27 controls, 27 HCV-RNA- HCV-exposed cases, 59 HCV-RNA+ patients with mild fibrosis and 48 HCV-RNA+ patients with severe fibrosis) by study group allocation. (D) Co-expression of γ-H2AX and phospho-p53 (Ser 15) in CD8+ T lymphocytes from subjects with viraemic HCV infection. (E and F) Co-expression of γ-H2AX and Ki67 in CD8+ T lymphocytes from 10 HCV-RNA+ subjects. Analysis by Wilcoxon signed rank test. (G) Cell-surface phenotype of circulating γ-H2AX + CD8+ T lymphocytes from subjects (n = 60) with HCV infection. CD8+ lymphocytes were divided based on the expression of the surface markers CD27 and CD57. Statistical analysis by Friedman test with Dunn’s multiple comparison test. (H) CD8+ T lymphocyte telomere length from CD8+ subsets based on CD27 expression from 10 HCV-RNA+ subjects. (I) Immunoblot of magnetic-bead separated CD8 + CD27+ and CD27- cells for γ-H2AX and β-actin. Irradiated whole CD8+ lymphocytes were a positive control for γ-H2AX expression.