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）
作者
王亚东
王玮
申川
张莉
商小波
梁乾飞
赵彩彦
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)
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2017年第1期5-10,共6页
Chinese Journal of Infectious Diseases
