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

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February 2002

The Immunization Study of Hepatitis C Virus E2 Gene DNA in Experimental Animal[J]. Virologica Sinica, 2002, 17(1): 11-16.

Citation: . The Immunization Study of Hepatitis C Virus E2 Gene DNA in Experimental Animal .VIROLOGICA SINICA, 2002, 17(1) : 11-16.

丙型肝炎病毒E2基因DNA免疫实验动物研究

1.
武汉大学生命科学学院病毒研究所．湖北武汉43OO72

摘要

抽要：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r~-DNA 3．1(一) 中的CMVIE启动子下游，掏建成HCVE2重组真棱表达质粒pcE2。EL1SA法检测pcE2DNA免疫兔血清中的融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抗体水平保持平德，抗体滴度达到L：1600左右。流式细胞计数仪(FACS)检测pcE2DNA免疫鼠CIM 、c瑚 T淋巴细胞变化情况．与注射空载体pCDNA3．1(一)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poE2的小鼠组织中有明显的阳性着色．而注射口cDNA3 1(一)的殖照组小鼠免疫组化结果为阴性以上结果表明：pcE2在宴验动物内表达出的HCVE2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。
- 丙型肝炎病毒(HCV)
- , DNA免疫
- , 免疫组化

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

1.
Institute of Virology，Wuhan University．Wuhan 430072，China

Abstract

Abstract：HCV E2 DNA fragment encoding E2 pmtein(amino acid 417—750)was cloned into the eu— caryotic expressing plasmid pCDNA3 to get the recombinant plasmid pcE2．The pcD2 DNA was used as gene vaccine to immunize rabbi~ and mice respectively．The specific antibody against HCV E2 was detected by ELISA at 20 day after immunization，and WaS raising to highest level at 40 day，then main· tained 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_ 1 increased 35 46％ aS measured by FACS．The immuuo-histo— chemicaI assay of the tissue from immune mice demostrated that the E2 gene had expressed inside the cells．this results indicated that E2 DNA has a good immunity
- Hepatitis C virus(Hcv)
- , DNA Immunization
- , Immunohistochemical asay

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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

1. Institute of Virology，Wuhan University．Wuhan 430072，China

Abstract: Abstract：HCV E2 DNA fragment encoding E2 pmtein(amino acid 417—750)was cloned into the eu— caryotic expressing plasmid pCDNA3 to get the recombinant plasmid pcE2．The pcD2 DNA was used as gene vaccine to immunize rabbi~ and mice respectively．The specific antibody against HCV E2 was detected by ELISA at 20 day after immunization，and WaS raising to highest level at 40 day，then main· tained 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_ 1 increased 35 46％ aS measured by FACS．The immuuo-histo— chemicaI assay of the tissue from immune mice demostrated that the E2 gene had expressed inside the cells．this results indicated that E2 DNA has a good immunity