丙型肝炎病毒E2基因DNA免疫实验动物研究  

The Immunization Study of Hepatitis C Virus E2 Gene DNA in Experimental Animal

作　　者：刘源洁[1] 叶林柏[1] 徐进平[1] 郜金荣[1] 冯磊[1] 孙明颢[1] 李江[1] 吴正辉[1] 

机构地区：[1]武汉大学生命科学学院病毒研究所,湖北武汉430072

出　　处：《中国病毒学》2002年第1期11-16,共6页Virologica Sinica

摘　　要：将编码丙型肝炎病毒（ＨＣＶ）Ｅ２蛋白４１７～７５０位氨基酸的ＤＮＡ片段　克隆到真核表达载体ｐｃＤＮＡ　３．１（－）中的ＣＭＶ　ＩＥ启动子下游，构建成ＨＣＶ　Ｅ２重组真核表达质粒　ｐｃＥ２。ＥＬＩＳＡ法检测ｐｃＥ２　ＤＮＡ免疫兔血清中的Ｅ２抗体变化和维持规律，结果显示免疫２０ｄ已有　抗体产生，３０ｄ后开始进入高峰，４０ｄ时达到最高值，至第９０ｄ抗体水平保持平稳，抗体滴度　达到１∶１６００左右。流式细胞计数仪（ＦＡＣＳ）检测ｐｃＥ２　ＤＮＡ免疫鼠ＣＤ４＋、ＣＤ８＋Ｔ淋巴细胞变　化情况，与注射空载体ｐＣＤＮＡ３．１（－）的阴性鼠相比，ＣＤ４＋淋巴细胞水平略有上升，ＣＤ８＋　细胞水平有较大升高，增幅达３５．４６％。免疫组化检测结果显示注射ｐｃＥ２的小鼠组织中有明显　的阳性着色，而注射ｐｃＤＮＡ３．１（－）的对照组小鼠免疫组化结果为阴性。以上结果表明：ｐｃＥ２　在实验动物内表达出的ＨＣＶ　Ｅ２蛋白可以引起免疫动物的体液免疫应答和细胞免疫应答，尤其　是ＭＨＣ－１限制性杀伤性ＣＤ８＋Ｔ淋巴细胞水平的提高对清除　病毒是十分有利的，因此ＨＣＶ　Ｅ２　ＤＮＡ免疫有可能成为预防和治疗ＨＣＶ感染的一条新途径。HCV E2 DNA fragment encoding E2 protein(amino acid 417-7 50) was cloned into the eucaryotic expressing plasmid pCDNA3 to get the recombin ant plasmid pcE2.The pcD2 DNA was used as gene vaccine to immunize rabbits and mice respectively.The specific antibody against HCV E2 was detected by ELISA at 20 th day after immunization,and was raising to highest level at 40th day ,then maintained at the titer of 1∶1 600.The CD4+ and CD8+ T-lymphocyte in immune mice increased compared with the control group,the CD8+ T-cell increa sed 35.46% as measured by FACS.The immuno-histochemical assay of the tissue fro m immune mice demostrated that the E2 gene had expressed inside the cells,this r esults indicated that E2 DNA has a good immunity.

关 键 词：丙型肝炎病毒 HCV DNA免疫 免疫组化 E2基因 实验动物 

分 类 号：R392[医药卫生—免疫学]