乙型肝炎病毒感染者单核细胞来源的树突状细胞中维甲酸诱导基因-I的表达及功能  

The expression and function of retinoic acid-inducible gene I in monocyte-derived dendritic ceils in patients with hepatitis B virus infection

作　　者：赵钢德[1] 谢青[1] 王晖[1] 安宝燕[1] 周慧娟[1] 贾妮娜[1] 林兰意[1] 石翠翠[1] 郭清[1] 俞红[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院感染科,200025

出　　处：《中华传染病杂志》2009年第12期727-732,共6页Chinese Journal of Infectious Diseases

基　　金：基金项目：国家自然科学基金资助项目（30671838）;十一五科技重大专项课题项目（2008ZX1002005、2008ZX1002-007）

摘　　要：目的探讨HBV感染后维甲酸诱导基因I（RIG-1）在单核细胞来源的树突状细胞（MoDC）中的表达和功能，并了解RIGI和MoDC在HBV感染疾病进程中的作用。方法采集28例HBV感染者外周血，21例为慢性乙型肝炎（CHB）患者，7例为急性乙型肝炎（AHB）患者；18例健康志愿者作为对照。采用CD14免疫微珠分离纯化外周血CD14^＋单核细胞，用粒-巨噬细胞集落刺激因子（GMCSF）和IL-4联合诱导培养7d成为树突状细胞（DC），用水泡性口炎病毒（VSV）感染刺激RIG-I活化。在不同时间点收集细胞抽提RNA，实时定量PCR检测RIG-I、干扰素启动子刺激基因1（IPS—1）和IFN-β的表达。正态分布数据采用方差分析，连续性变量两组间比较采用Mann—Whitney U检验，多组间比较采用Kruskal—Wallis检验。结果VSV感染活化RIG-I后，CHB、AHB组和健康对照组的RIG1mRNA表达水平在16h（2．44±2．03，19．54±3．15，21．48±8．39；F=7．451，P=0．002）和24h（2．68±2．93，10．31±3．88，14．01±5．04；F=7．908，P=0．001）时差异均有统计学意义；IPS-1在CHB和AHB组中的表达在16h（2．05±1．08，1．99±1．56）和24h（2．27±2．16，3．24±1．21）均高于健康对照组（16h：0．60±0．31，F=7，246，P=0．003；24h：1．08±0．73，F=13．598，P=0．01）。CHB组中IFN-β水平在16h和24h均低于AHB组和健康对照组，差异有统计学意义。结论RIG-I和IPS-1在HBV感染者MoDC中的表达明显异常，提示RIGI信号通路可能被HBV干扰，RIG—I和MoDC的功能缺陷在HBV感染和慢性化中发挥重要作用。Objective To investigate the expression and function of retinoic acid inducible gene I （RIG-I） in monocyte derived dendritic ceils （MoDC） at different stages of hepatitis B virus（HBV） infection and to explore the role of RIG-I in the disease progression after HBV infection. Methods Peripheral blood samples were collected from 28 hepatitis B virus infected persons, including 21 cases of chronic hepatitis B （CHB） and 7 of acute hepatitis B （AHB）. Eighteen healthy subjects were recruited as controls. Purified CD14^＋ monocytes were isolated by CD14 microbeads. MoDCs were induced from CD14^- rnonoeytes with granulocyte macrophage-colony stimulating factor （GM CSF） and interleukin （IL）-4 for 7 days, and then were infected with vesicular stomatitis virus （VSV） to stimulate RIG-I expression. The mRNA expression levels of RIG-I, interferon （IFN）-promoter stimulating factor-1 （IPS 1） and IFN-β at 16 hours and 24 hours after infection with VSV were measured by real-time quantitative polymerase chain reaction （PCR）. Data with normal distribution were tested by analysis of variance. Continuous variables between groups were compared using Mann Whitney U test. Comparison among multiple groups was clone by Kruskal-Wallis test. Results The expression levels of RIG-I in MoDCs from CHB patients were significantly lower than those in AHB patients and healthy controls at 16 hours （2.44±2.03, 19.54±3.15, 21.48±8.39, respectively; F=7. 451,P= 0.002） and 24 hours （2.68±2.93, 10.31± 3. 88, 14.01±5.04, respectively; F=7.908,P=0.001） following VSV stimulation. The IPS-1 levels in both CHB patients and AHB patients were higher than those in healthy controls at 16 hours （2.05± 1.08, 1.99 ± 1.56, 0.60 ± 0. 31, respectively; F= 7. 246, P =0.003） and24 hours （2. 27±2. 16, 3.24±1.21, 1.08±0.73, respectively; F=13. 598,P=0.001）. Furthermore, the IFN-β expression levels were significantly lower in CHB patients compared to AHB patients and healthy controls at 16 hours and 2

关 键 词：肝炎 乙型 慢性 树突细胞 维甲酸 基因疗法 干扰素类 免疫 天然 

分 类 号：R512.62[医药卫生—内科学]