Fig. 2: Glucose metabolism is impaired in HCV-specific CD8+ T cells from chronically evolving acute patients.

a Representative examples of virus-specific CD8+ T cells stained with HLA-A2+ dextramers ex vivo after overnight anti-CD3/anti-CD28 stimulation. Glucose uptake (b), measured by the incorporation of the glucose analog 2-NBDG (MFI), and Glut1 expression levels (c) in virus-specific CD8+ T cells from T1/early HCV patients and healthy controls stimulated as in a. Data are presented as median fluorescence intensity (MFI) values; median values are indicated by horizontal lines. Different numbers of patients (represented by individual dots) were tested in each assay depending on dextramer-positive cell frequencies. b–c Differences between multiple groups were evaluated with the non-parametric Kruskal-Wallis test; p-values were corrected for pairwise multiple comparisons with the Dunn’s test. d metabolic flux profiling of purified CD8+ T cells from 6 T1/early chronically-evolving (acute) patients. Cells were stimulated overnight with either HCV-NS3 (red) or control (FLU-specific, CMV-specific and EBV-specific) peptides (blue), or were not stimulated (green). The extracellular acidification rate (ECAR) was measured in real-time ex vivo before (basal level) and after oligomycin treatment in order to determine the maximum glycolytic capacity (MGC) and glycolytic reserve (difference between MGC and baseline ECAR) (see Methods section for details on Seahorse analysis). e Metabolic flux profiling of purified CD8+ T cells from T1/early self-limited (acute) patients (n = 4) stimulated overnight as in d. ECAR, maximum glycolytic capacity and glycolytic reserve were measured as in d. In d and e, ECAR values are given as the mean ± SD in the left-side and are presented as box-and-whisker plots (with median and 5–95 percentile) in the right-side. d–e Statistical analysis was performed with the Friedman test to compare different stimuli; p-values have been corrected for pair-wise multiple comparisons with the Conover’s test.