HBeAg特异性细胞免疫反应体外抗乙肝病毒作用  

Anti-HBV effect of HBeAg specific cellular immune response in vitro

作　　者：陈瑾[1] 吴金明[2] 林贤凡[2] 孙慧[2] 金思思[2] 申苏建[2] 

机构地区：[1]台州市立医院儿科,浙江台州318000 [2]温州医学院附属第一医院消化内科

出　　处：《胃肠病学和肝病学杂志》2010年第4期313-316,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：温州市2009年科技计划项目(Y20090036)

摘　　要：目的探讨体外HBeAg特异性细胞免疫反应对乙型肝炎病毒(HBV)mRNA表达及蛋白合成的影响。方法自正常人外周血单核细胞中诱导培养树突状细胞(DC),经不同浓度的HBeAg负载后与自身T细胞共同培养,诱导产生HBeAg特异性T细胞,同时设与无HBeAg刺激的DC共同培养的T细胞、单纯T细胞为对照组,作用于HepG2.2.15细胞系,逆转录PCR(RT-PCR)检测HepG2.2.15细胞内乙肝表面抗原(HBsAg)、乙肝核心抗原(HBcAg)的mRNA水平,酶联免疫法(ELISA)检测细胞培养上清中HB-sAg和乙肝e抗原(HBeAg)水平。结果经HBeAg刺激激活的T细胞可以有效地抑制HBsAg、HBcAg mRNA的表达(P<0.05)及HBsAg、HBeAg的合成(P<0.05),且随着HBeAg刺激浓度的升高,对上述指标的抑制率逐渐上升。结论HBeAg特异性细胞免疫反应可显著抑制HBV mRNA和蛋白的表达,为开发HBeAg相关的治疗性疫苗奠定理论基础。Objective To investigate the effect of HBeAg specific cellular immune response on hepatitis B virus mRNA and protein expression in vitro.Methods The monocyte-derived dendritic cells（DC） were generated from healthy people and cocultured with self-blood T lymphocyte after loaded with different concentration of HBeAg to induce specific T lymphocyte.While DC without HBeAg loaded group,only T lymphocyte group were established as control group.HepG2.2.15 cells and culture supernatant were collected after stimulated with T lymphocyte.Intracellular hepatitis B surface antigen（HBsAg） and hepatitis B core antigen（HBcAg） mRNA were analyzed by reverse transcription-PCR（RT-PCR）.HBsAg and hepatitis B e antigen（HBeAg） in supernatant were detected by enzyme-linked immunosorbent assay（ELISA）.Results Antigen-activated T lymphocyte could suppress the expressions of HBsAg,HBcAg mRNA（P〈0.05） and decrease the levels of HBsAg,HBeAg（P〈0.05） in a dose-dependant manner.Conclusion HBeAg specific cellular immune response can inhibit the expressions of HBV mRNA and protein.This will lay theoretical foundation for developing therapeutic vaccine about HBeAg.

关 键 词：乙型肝炎病毒 树突状细胞 细胞免疫 HEPG2.2.15细胞 

分 类 号：R512.62[医药卫生—内科学]