丙型肝炎病毒多表位基因载体的构建与表达  

Construction and expression of recombined plasmids carrying genes of the hepatitis C virus that encode multiple epitopes

作　　者：邝玉[1] 孙梦雯[1] 杨远[1] 马巨辉[1] 李明远[1] 

机构地区：[1]四川大学基础医学与法医学院微生物学教研室,四川成都610041

出　　处：《中国病原生物学杂志》2015年第4期293-298,共6页Journal of Pathogen Biology

基　　金：四川大学实验技术立项资助课题(No.0083304116008)

摘　　要：目的构建编码多个丙型肝炎病毒(HCV)抗原表位的真核表达载体,建立稳定转染的细胞株,并利用小鼠移植瘤模型初步评估疫苗在动物体内的免疫反应状况。方法合成含10条CTL细胞表位的M2基因,与含多条B细胞及T细胞表位的M1基因连接合成M1M2基因;将M2、M1M2基因分别与pcDNA3.1(+)连接,构建真核表达载体pcDNA3.1(+)/M2及pcDNA3.1(+)/M1M2;将上述两种载体与pcDNA3.1(+)/M1分别转入SP2/0细胞中,并检测目的蛋白表达;用3种重组质粒分别免疫BALB/c小鼠移植瘤模型,初步评估疫苗引起的免疫反应。结果 3种真核表达载体在SP2/0细胞中均能得到稳定表达,免疫荷瘤小鼠后,小鼠瘤体的大小M1M2组<M2组<M1组<空质粒对照组,实验组均能观察到肿瘤组织的坏死现象。结论成功构建了HCV多表位基因的真核表达载体pcDNA3.1(+)/M1M2,用该表达载体免疫小鼠可引起较强的特异性免疫应答。Objectives To construct vectors for eukaryotic expression of genes of the hepatitis C virus （HCV） that en- code multiple epitopes, to establish stable cell lines by transfecting those vectors into SP2/0 cells, and to attempt to eval- uate the immune reaction induced by a vaccine in a mouse xenograft tumor model. Methods The gene M2, which in cludes 10 epitopes of CTL, was synthesized, and the gene M1M2 was synthesized by combining M2 with M1, which includes various B cell and T cell epitopes. The genes M2 and M1 M2 were obtained and inserted into pcDNA3.1 （ ＋ ） to con- struct the eukaryotic expression vectors pcDNA3.1 （＋）M2 and pcDNA3.1 （＋）M1 M2. SP2/0 cells were transfected with the three recombinant plasmids （pcDNA3.1 （ ＋ ） M1 , pcONA3.1 （ ＋ ） M2 , and pcDNA3.1 （ ＋ ） M1 M2 ） , and the expression of HCV proteins with multiple epitopes was determined. BALB/c mice bearing xenografted tumors were immunized with the three recombinant plasmids and their immune response was observed. Results The three recombinant plasmids were stably expressed in SP2/0 cells and tumor-bearing mice. Substantial tumor suppression was noted in all of the immunized mice. Conclusion Vectors for eukaryotic expression of genes of the HCV that encode multiple epitopes were successfully constructed, and the pcDNA3.1 （＋）M1 M2 plasmid induced a specific immune response in mice.

关 键 词：丙型肝炎病毒 多表位基因 核酸疫苗 

分 类 号：R373.21[医药卫生—病原生物学]