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Additional Supporting Information may be found in the online version of this article.

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HEP_23977_sm_SuppInfoFigure1.tif7836KSupporting Figure 1. The quantitative analysis of CD56+ (A) and CD3+ (B) cells in both portal and lobular areas in the liver tissues of these subjects. Each dot represents one individual. The horizontal bars indicate the median percentiles. The significant P values are shown. hpf, high-power field.
HEP_23977_sm_SuppInfoFigure2.tif5634KSupporting Figure 2. The gating strategy of intrahepatic NK cells. Gate 1 was used to select CD45+ cells and to exclude red blood cells and cell debris. Gate 2 was used to separate liver-infiltrating lymphocytes from other cells (excluding monocytes and granulocytes) by forward and side scatter. Finally, the cell subsets including CD3-CD56+ NK cells (left-upper quadrants), CD3+CD56+ NKT cells (right-upper quadrants) and CD3+CD56− T cells (right-lower quadrants) were selected.
HEP_23977_sm_SuppInfoFigure3.tif17529KSupporting Figure 3. Representative dot plots depict the expression of activation receptors (NKp30, NKp44, NKp46 and NKG2D) and inhibitory receptors (CD158a, CD158b and NKG2A) in NK cells from LILs and PBMCs in the three groups. CD3−CD56+ NK cells were gated. Values in the upper-right quadrants represent the percentages of CD3−CD56+ NK cells that express NK receptors. LILs, liver-infiltrating lymphocytes; PBMCs, peripheral blood mononuclear cells; HC, healthy controls; IT, immune tolerant subjects with chronic HBV infection; IA, immune activated patients with chronic HBV infection.
HEP_23977_sm_SuppInfoFigure4.tif12407KSupporting Figure 4. The spontaneous degranulation of NK cells was significantly increased in IA patients. (A) Representative dot plots depict the spontaneous degranulation of both intrahepatic and peripheral NK cells among IA patients and IT and HC subjects. CD3−CD56+ NK cells were gated. Values in the upper-right quadrants represent the percentages of CD3−CD56+ NK cells that express CD107a. (B) Pooled data show the percentages of intrahepatic and peripheral CD3−CD56+ NK cells that express CD107a from the three groups. Each dot represents one individual. The horizontal bars indicate the median percentiles. Multiple comparisons were made using the Kruskal-Wallis H non-parametric test among the different groups. The Mann-Whitney U test was used to compare data from two different groups. The significant P values are shown.
HEP_23977_sm_SuppInfoFigure5.tif6679KSupporting Figure 5. NKp30 predominantly accounted for the NCR-mediated NK cell degranulation seen in IA patients. The redirected assay of NK cells against P815 target cells using mixed anti-NCR Abs or individual anti-NKp30, NKp44 and NKp46 antibodies was performed. Pooled data show the percentages of NK cells expressing IFN-? (A) and CD107a (B) in the redirected assay. The Wilcoxon matched pairs T test was used to compare the data from same individuals. *P < 0.05.
HEP_23977_sm_SuppInfoFigure6.tif10694KSupporting Figure 6. Hepatic mRNA expression analysis of NK receptor ligands in IA patients and HC and IT subjects. Statistical analysis of the mRNA expression of the ligands of NK receptors in the liver, including the NKG2D ligands, MICa, MICb, ULBP1, 2, 3 and 4, the NKG2A ligand, HLA-E, and NKp30 ligands, BAT3 and B7H6, from IA patients (n = 10) and IT (n = 6) and HC subjects (n = 7). The Mann-Whitney U test was used to compare data from two different groups. The P values are shown.
HEP_23977_sm_SuppInfo.doc106KSupporting Information

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