Distribution of Hepatitis C virus (HCV) genotypes in studied samples (n = 182).

<p>Distribution of Hepatitis C virus (HCV) genotypes in studied samples (n = 182).</p

Characteristics of the commercially available HCV genotyping assays.

<p>Characteristics of the commercially available HCV genotyping assays.</p

Genotyping results with Roche HCV genotyping kit on samples with low HCV viral load.

<p>(p = 0.006) Successes: filled circles; failures: filled squares; (p = 0.006).</p

Correspondence between results obtained with both techniques for the regular sample panel (n = 101).

<p>Correspondence between results obtained with both techniques for the regular sample panel (n = 101).</p

Correspondence between results obtained with both techniques for the low viral load samples (n = 42).

<p>Correspondence between results obtained with both techniques for the low viral load samples (n = 42).</p

Patient 3 HCV alignments

6 HCV sequence alignments for patient 3 - Core and NS5B regions, 3 time sample

Patient 2 HVC alignments

10 HCV sequence alignments for patient 2 - Core and NS5B regions, 5 time samples

Direct HBV-specific CD8 T-cell response evolution during Peg-IFN-α therapy.

<p>Direct HBV-specific CD8 T cells measurements following Peg-IFN-α treatment using tetramers. (A) Representative dot plots of the tetramer labeling of HBc_18-27- and HBs_335-343-specific T cells (gated on CD8 T cells) before and 24 weeks after treatment in patients treated with nucleos(t)ide analog alone (<i>left panel</i>) or together with Peg-IFN-α (<i>right panel</i>). (B) Evolution of the basal percentages of HBc_18-27- (left panel) and HBs_335-343- (right panel) specific CD8 T cells in patients with CHB infection treated with nucleos(t)ide analog alone (<i>white bars</i>, n = 7) or together with Peg-IFN-α (<i>grey bars</i>, n = 7). The gray area represents the period of Peg-IFN-α administration.</p

B-cell subsets during Peg-IFN-α therapy.

<p>(A) B-cell gating strategy. The peripheral B-cell subsets were classified according to the most common lineage/differentiation markers CD19, CD10, CD27, CD38, IgD, and IgM. B-cell subsets were defined as: transitional B cells (CD19⁺CD27^-IgD⁺CD10⁺CD38^high); naive B cells (CD19⁺CD27^-IgD⁺CD10^-CD38^low); natural memory B cells (CD19⁺CD27⁺IgD⁺); post-GC memory B cells (CD19⁺CD27⁺IgD^-CD10^-CD38^low); plasmablasts (CD19⁺CD27⁺IgD^-CD10^- CD38^high); CD27^-IgD^- memory B cells (CD19⁺CD27^-IgD^-) and activated B cells (CD19⁺CD27⁺IgD^-CD10⁺CD38^low). Post-GC memory B cells were further subdivided into IgM⁺ and IgM^- switched B cells. Representative dotplots from one patient with CHB infection. (B) Modulation of total B cells by Peg-IFN-α. Frequency (within PBMC) and absolute numbers of total B cells in patients with CHB infection treated with nucleos(t)ide analog alone (<i>open circles</i>, n = 11–14) or together with Peg-IFN-α (<i>black circles</i>, n = 8–9). The gray area represents the period of Peg-IFN-α administration. Bars represent median. <i>P</i> values were calculated using the Wilcoxon test (<i>straight lines</i>) or the Mann-Whitney test (<i>dashed lines</i>). * p<0.05, ** p<0.01, *** p<0.001.</p

Functions of Liver Natural Killer Cells Are Dependent on the Severity of Liver Inflammation and Fibrosis in Chronic Hepatitis C

<div><p>During chronic hepatitis C virus (HCV) infection, the role of intra-hepatic (IH) natural killer (NK) cells is still controversial. To clarify their functions, we investigated anti-viral and cytotoxic activity of NK cells in human fresh liver biopsies. We compared the functions of IH-NK cells in HCV-infected and NASH patients in physiological conditions as well as after stimulation using flow cytometric and immunohistochemical analyses. Interestingly, few IH-NK cells produced anti-viral cytokine IFN-γ in HCV-infected patients similarly as in non-infected individuals. Spontaneous degranulation activity was extremely low in peripheral NK cells compared to IH-NK cells, and was significantly higher in IH-NK cells from HCV-infected patients compared to non-infected individuals. Immunohistochemical analysis revealed that perforin granules were polarized at the apical pole of IH-NK cells. The presence of CD107a and perforin in IH-NK cells demonstrated that NK cells exerted a cytolytic activity at the site of infection. Importantly, IH-NK cell functions from HCV-infected patients were inducible by specific exogenous stimulations. Upon <i>ex vivo</i> K562 target cell stimulations, the number of degranulating NK cells was significantly increased in the pool of IH-NK cells compared to circulating NK cells. Interestingly, after stimulation, the frequency of IFN-γ-producing IH-NK cells in HCV-infected patients was significantly higher at early stage of inflammation whereas the spontaneous IH-NK cell degranulation activity was significantly impaired in patients with highest inflammation and fibrosis Metavir scores. Our study highlights that some IH-NK cells in HCV-infected patients are able to produce INF-γ and degranulate and that those two activities depend on liver environment including the severity of liver injury. Thus, we conclude that critical roles of IH-NK cells have to be taken into account in the course of the liver pathogenesis associated to chronic HCV infection.</p></div