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Supplementary material for this article can be found on the H EPATOLOGY website ( http://interscience.wiley.com/jpages/0270-9139/suppmat/index.html ).

FilenameFormatSizeDescription
jws-hep.21438.fig1A.pdf17KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig2B.pdf61KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1C.pdf32KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1D.pdf66KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1E.pdf22KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1F.pdf69KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1G.pdf23KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig1H.pdf60KFigure S1:Detection of intrahepatic HCV-specific CD8 T lymphocytes using confocal microscopy. Merged colors: CD8 (red), HCV-specific CD8 (green) and an overlay with DIC image are shown for a single HCV-tetramer staining:a,b: NS3-2 (1406); c,d: Core (41); e,f: Core (35); g,h: NS5B (2727).
jws-hep.21438.fig2A.pdf77KFigure S2:Detection of HCV-specific CD8 T lymphocytes in peripheral blood. a:Gating strategy used. Lymphocytes were gated on CD8high cells to exclude NK cells and monocytes, dead cells were excluded using 7AAD staining, and two HCV-tetramer (PE-labeled and APC-labeled) dot-plot was used to eliminate double positive cells. To discriminate the low fluorescence intensity events, regions of tetramer positivity (R4 and R5) were defined on a control sample and applied to the patient sample analysis. b:Examples of HCV-tetramer staining on PBMC. Numbers indicate the percentage of CD8highcells positive for HCV-tetramer staining. c:Tetramer staining of polyclonally-expanded intrahepatic lymphocytes (IHL). IHL isolated from liver biopsies of HCV-infected patients were stimulated with PHA, IL2 and autologous PBMC for 2 weeks. Dot-plots show tetramer staining on CD8high 7-AAD(-) lymphocytes.
jws-hep.21438.fig2C.pdf70KFigure S2:Detection of HCV-specific CD8 T lymphocytes in peripheral blood. a:Gating strategy used. Lymphocytes were gated on CD8high cells to exclude NK cells and monocytes, dead cells were excluded using 7AAD staining, and two HCV-tetramer (PE-labeled and APC-labeled) dot-plot was used to eliminate double positive cells. To discriminate the low fluorescence intensity events, regions of tetramer positivity (R4 and R5) were defined on a control sample and applied to the patient sample analysis. b:Examples of HCV-tetramer staining on PBMC. Numbers indicate the percentage of CD8highcells positive for HCV-tetramer staining. c:Tetramer staining of polyclonally-expanded intrahepatic lymphocytes (IHL). IHL isolated from liver biopsies of HCV-infected patients were stimulated with PHA, IL2 and autologous PBMC for 2 weeks. Dot-plots show tetramer staining on CD8high 7-AAD(-) lymphocytes.
jws-hep.21438.fig3A.pdf18KFigure S3:Examples of TUNEL staining (CD8 - red, tetramer+CD8+ - yellow, TUNEL ? blue).a:NS5N (2595)-tetramer staining;b:merged image tetramer Core 132 and DIC picture. Few TUNEL+ cells were observed in the vicinity of tetramer-positive cells.
jws-hep.21438.fig3B.pdf54KFigure S3:Examples of TUNEL staining (CD8 - red, tetramer+CD8+ - yellow, TUNEL ? blue).a:NS5N (2595)-tetramer staining;b:merged image tetramer Core 132 and DIC picture. Few TUNEL+ cells were observed in the vicinity of tetramer-positive cells.
jws-hep.21438.fig4A.pdf15KFigure S4:Examples of laminin staining.a-c:low laminin expression in the regions containing tetramer+ cells: NS3-1 (1073) HCV-tetramer(a) , NS3-2 (1406)(b) , NS4B (1807)(c) .d:High laminin expression in a region of liver without any tetramer+ cell.
jws-hep.21438.fig4B.pdf17KFigure S4:Examples of laminin staining.a-c:low laminin expression in the regions containing tetramer+ cells: NS3-1 (1073) HCV-tetramer(a) , NS3-2 (1406)(b) , NS4B (1807)(c) .d:High laminin expression in a region of liver without any tetramer+ cell.
jws-hep.21438.fig4C.pdf16KFigure S4:Examples of laminin staining.a-c:low laminin expression in the regions containing tetramer+ cells: NS3-1 (1073) HCV-tetramer(a) , NS3-2 (1406)(b) , NS4B (1807)(c) .d:High laminin expression in a region of liver without any tetramer+ cell.
jws-hep.21438.fig4D.pdf24KFigure S4:Examples of laminin staining.a-c:low laminin expression in the regions containing tetramer+ cells: NS3-1 (1073) HCV-tetramer(a) , NS3-2 (1406)(b) , NS4B (1807)(c) .d:High laminin expression in a region of liver without any tetramer+ cell.

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