慢性乙型肝炎病毒感染者不同免疫状态下自然杀伤细胞G2D表达及其活化意义  

Significance of natural killer cell G2D expression and activation in patients with different immune status of chronic hepatitis B virus infection

作　　者：王亚东[1] 王玮[1] 申川[1] 张莉[1] 商小波 梁乾飞 赵彩彦[1] Wang Yadong Wang Wei Shen Chuan Zhang Li Shang Xiaobo Liang Qian fei Zhao Caiyan.(Department of Infectious Diseases, the Third Affiliated Hospital of Hebei Medical University, Shijiazhuang 050051 , China)

机构地区：[1]河北医科大学第三医院感染科,石家庄050051

出　　处：《中华传染病杂志》2017年第1期5-10,共6页Chinese Journal of Infectious Diseases

摘　　要：目的观察慢性HBV感染者不同免疫状态下NK细胞G2D（NKG2D）表达及其活化差异，探讨NKG2D在介导HBV感染后机体免疫炎性损伤过程中的意义。方法2010年1月至2011年12月河北医科大学第三医院收治慢性HBV携带者（免疫耐受）、CHB患者（免疫活动）及HBV相关慢加急性（亚急性）肝功能衰竭（HBV-ACLF）患者（免疫过强）各15例，流式细胞学检测PBMC中NK及NKG2D＋ NK细胞频率，实时荧光定量PCR检测肝组织NKG2D mRNA表达，免疫组织化学染色观察肝组织NKG2D＋细胞数量与分布，ELISA法检测血清γ干扰素、TNF-α、穿孔素、颗粒酶B水平。正态分布且方差齐时，采用单因素方差分析，组间比较采用SNK-q检验。非正态分布或方差不齐时，采用非参数Kruskal-Wallis H秩和检验，组间比较采用Nemenyi检验。分类变量采用Pearson χ^2检验。结果健康对照组外周血PBMC中NK细胞频率为（13.58±3.24）%，慢性HBV携带者组为（5.42±2.18）%，HBV-ACLF组为（7.92±2.85）%，CHB组为（8.43±2.92）%，健康对照组与慢性HBV携带者组、HBV-ACLF组、CHB组比较差异均有统计学意义（F＝22.04，P〈0.05）。其中NKG2D＋ NK细胞占全部NK细胞比例，以HBV-ACLF组患者最高，为（18.92±5.85）%，CHB组为（12.85±3.39）%、健康对照组为（8.45±2.86）%，慢性HBV携带者组为（3.36±1.05）%，各组间比较差异均具有统计学意义（H＝46.09，P〈0.01）。HBV-ACLF患者肝组织NKG2D mRNA与NKG2D＋细胞表达分别为6.58±1.86和30.69±6.67，CHB组为3.25±0.95和17.36±4.13，慢性HBV携带者组为0.69±0.20和3.16±1.24，各组间比较差异均具有统计学意义（H值分别为52.10和52.73，均P〈0.01）。血清γ干扰素、TNF-α、穿孔素、颗粒酶B水平变化趋势与肝组织NKG2D变化一致。结论慢性HBV感染者不同免疫状态NKG2D表达存在差异，NKG2D活化参与HBV感染后免疫病理损伤过程。Objective To investigate the differences of expression and activation of natural killer （NK） cell G2D （NKG2D） in patients with different immune status of chronic hepatitis B virus （HBV） infection, and to explore the significance of NKG2D-mediated immune injury in HBV infection. Methods Fifteen chronic HBV carriers （immune tolerance）, 15 chronic hepatitis B （CHB, immune activation） patients, 15 HBV-related acute/subacute-on-chronic liver failure （HBV-ACLF, immune over-activation） patients were enrolled in this study from January 2010 to December 2011 in the Third Hospital of Hebei Medical University. The frequencies of NK cells and NKG2D＋ NK cells in peripheral blood mononuclear cells （PBMC） were detected by flow cytometry. The NKG2D mRNA expressions were measured by real- time fluorescent quantitative polymerase chain reaction. Localization and hemi-quantitative analysis of NKG2D＋ cells in liver tissue were performed by immunohistochemistry staining. Concentrations of serum interferon（IFN）-γ, tumor necrosis factor（TNF）-α, perforin and granzyme B were quantified by enzyme linked immunosorbent assay （ELISA）. Normally distributed continuous variabIes were analyzed using one-way analysis of variance （ANOVA）, followed by Student-Newman-Keuls q test for evaluating variances between each two groups. For non-normally distributed data or heterogeneity of variance, differences between groups were analyzed using nonparametric Kruskal-Wallis H test, followed by Nemenyi test for pairwise comparisons. Pearson chi-square test was used to analyze categorical variables. Results The percentages of NK cells in PBMC were （13. 58 ± 3. 24）% in healthy controls, （5. 42± 2.18）% in chronic HBV carriers, （7.92±2.85）% in HBV-ACLF group and （8.43±2. 92）% in CHB group. The percentage of NK cells in PBMCs was lower in each chronic HBV-infected group compared with healthy controls （F=22.04, P〈0.05）. The frequency of NKG2D＋ NK ceils in HBV-ACLF group （18.92±5.85）

关 键 词：杀伤细胞 天然 NKG2D 肝炎病毒 乙型 肝炎 乙型 慢性 慢加急性肝衰竭 

分 类 号：R512.62[医药卫生—内科学]