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乙型肝炎病毒复制调控元件对HBV DNA疫苗诱导的免疫应答
引用本文:赫兢,辛绍杰,毛远丽,王海滨,周讯,杨健洋,孔维,貌盼勇. 乙型肝炎病毒复制调控元件对HBV DNA疫苗诱导的免疫应答[J]. 中华传染病杂志, 2006, 24(5): 311-315
作者姓名:赫兢  辛绍杰  毛远丽  王海滨  周讯  杨健洋  孔维  貌盼勇
作者单位:100039,北京,解放军第三○二医院传染病研究所病毒研究
摘    要:目的研究乙型肝炎病毒(HBV)复制调控元件增强子Ⅰ(ENHⅠ)及前S2(Pre-S2)抗原基因对HBV DNA疫苗诱导的免疫应答的影响。方法采用常规聚合酶链反应(PCR)从HBV adr亚型全基因DNA序列中分别扩增HBsAg、PreS2-HBsAg、HBsAg-ENHI和PreS2-HBsAg-ENHⅠ基因片段,重组到VR1012载体中,构建4种HBV DNA疫苗,转染HepG2细胞并免疫Balb/C小鼠。通过细胞免疫化学、酶联免疫分析(ELISA)、酶联免疫斑点试验(ELISPOT)等方法检测其在HepG2细胞内的表达及小鼠的体液及细胞免疫应答。结果转染的HepG2细胞表达相应的目的蛋白.ENHⅠ及Pre-S2抗原基因均可增强HBV DNA疫苗转染HepG2细胞表达HBsAg;免疫接种小鼠后第2周产生抗-HBs及HBsAg特异性细胞毒T淋巴细胞(CTL),Pre—S2抗原基因可增强HBV DNA疫苗免疫Balb/C小鼠诱导的抗-HBs及HBsAg特异性CTL的产生,ENHⅠ基因对免疫应答无影响。结论ENHI及Pre—s2抗原基因均可增强HBVDNA疫苗转染HepG2细胞表达HBsAg.Pre-S2抗原基因可增强HBVDNA疫苗免疫Balb/C小鼠引起的免疫应答。

关 键 词:肝炎  乙型 疫苗 DNA 肝炎抗原  乙型 增强子元件(遗传学)
收稿时间:2005-11-15
修稿时间:2005-11-15

The influence of HBV replication regulator on the immune response induced by HBV DNA vaccine
HE Jing, XIN Shao-jie, MAO Yuan-li, et al. The influence of HBV replication regulator on the immune response induced by HBV DNA vaccine[J]. Chinese Journal of Infectious Diseases, 2006, 24(5): 311-315
Authors:HE Jing   XIN Shao-jie   MAO Yuan-li   et al
Affiliation:Department of Virology, Institute of Infectious Disease,302 Hospital of PLA , Beijing 100039, China
Abstract:Objective To study the influence of HBV replication regulator,enhancer I and Pre- S2,on the immune response of HBV DNA vaccine.Methods DNA fragments of HBsAg,PreS2 HBsAg,HBsAg-enhancer I and PreS2-HBsAg-enhancer I region of HBV were amplified by PCR using the complete genome DNA of HBV adr subtype,and inserted into VR1012 vectors,respective- ly.The recombinant plasmids were transfected into HepG2 cells,and injected into Balb/C mice.The expression of HepG2 cells and the cellular and humoral immune response of mice were tested by cell immuno-chemistry,ELISA and ELISPOT.Results The target protein were expressed by transfected HepG2 cells,enhancer I and Pre-S2 can promote the expression of HBsAg in transfected cells.The HBsAb and the HBsAg specific CTL in inoculated mice were found in the second week after injection, PreS2 but not enhancer I can promote the immune response in inoculated mice.Conclusions When inserted into HBV DNA vaccine,enhancer I and PreS2 can promote the expression of HBsAg in transfected HepG2 cells,PreS2 can promote the immune response in inoculated Balb/C mice.
Keywords:Hepatitis B  Vaccine  DNA  Hepatitis B antigens  Enhancer elements(genetics)
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