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颗粒化重组戊型肝炎病毒衣壳蛋白及其抗原性与免疫原性
引用本文:何志强,张军,李少伟,林鉴,刘如石,陈毅歆,王颖彬,夏宁邵.颗粒化重组戊型肝炎病毒衣壳蛋白及其抗原性与免疫原性[J].生物工程学报,2004,20(2):262-268.
作者姓名:何志强  张军  李少伟  林鉴  刘如石  陈毅歆  王颖彬  夏宁邵
作者单位:厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,福建省医学分子病毒学研究中心,厦门,361005
基金项目:福建省科技重大项目 (No .2 0 0 2F0 1 3),教育部跨世纪优秀人才培养计划~~
摘    要:过去的研究发现大肠杆菌表达的戊型肝炎病毒(HEV)衣壳蛋白ORF2的aa394-606片段NE2可以形成同源多聚体,并具有良好的免疫保护性,但纯化后的免疫原性较弱。这里表达了3个NE2蛋白的N端延伸突变体,发现对应于ORF2 aa368_606的重组蛋白HEV239在体外可以形成颗粒性抗原。HEV 239抗原颗粒与戊肝患者血清反应性良好,对中和性单克隆抗体8C11的反应性与NE2抗原相当,而对另一中和性单克隆抗体8H3的反应性较NE2抗原有显著提高,表明HEV 239抗原颗粒具有比NE2更好的抗原性。纯化后的HEV 239抗原颗粒直径约为15~30nm。铝佐剂吸附的HEV 239免疫Balb/c小鼠的半数有效剂量(ED50)在0.08~0.25μg之间,而同样以铝佐剂吸附的NE2抗原60μg剂量免疫的抗体阳转率仅25%,表明HEV 239抗原颗粒具有更好的免疫原性。

关 键 词:戊型肝炎病毒,  衣壳蛋白,  疫苗,  颗粒性抗原,  原核表达
文章编号:1000-3061(2004)02-0262-07
修稿时间:2003年8月15日

Particulate Recombinant Hepatitis E Virus Capsid Protein and Its Antigenicity and Immunogenicity
HE Zhi_Qiang\ ZHANG Jun\ LI Shao_Wei\ LIN Jian\ LIU Ru_ShiCHEN Yi_Xin\ WANG Ying_Bin\ XIA Ning_Shao.Particulate Recombinant Hepatitis E Virus Capsid Protein and Its Antigenicity and Immunogenicity[J].Chinese Journal of Biotechnology,2004,20(2):262-268.
Authors:HE Zhi_Qiang\ ZHANG Jun\ LI Shao_Wei\ LIN Jian\ LIU Ru_ShiCHEN Yi_Xin\ WANG Ying_Bin\ XIA Ning_Shao
Institution:The Key Laboratory of Ministry of Educationfor Cell Biology and Tumor Cell Engineering, Research Center for Medical molecular virology of Fujian Province, Xiamen University, Xiamen 361005, China.
Abstract:An E. coli expressed recombinant antigen NE2 was reported to aggregate into homo-oligomer, and can induce protective antibodies on rhesus monkey, but its immunogenicty was much weak after being purified. In this study, three N-terminal extension mutant of NE2 were expressed in E. coli, one of which named HEV 239 was found to aggregate into particle. HEV 239 antigen had good reactivity with sera of hepatitis E patients. The reactivity of HEV 239 against neutralization monoclonal antibody 8C11 was similar as NE2 antigen, while the reactivity of it against another neutralization monoclonal antibody 8H3 is much better than NE2 antigen, which indicated better antigenicity of HEV 239 than NE2. The diameter of purified HEV 239 particulate antigen was between 15 nm to 30 nm. The ED50 of immunization of HEV 239 particle adsorbed by aluminum adjuvant to BALB/c mice was between 0.08 microg to 0.25 microg. In contrast, the seraconversion rate of mice immunized by NE2 antigen adsorbed by aluminium adjuvant was only 25% on 60 microg vaccination. These results suggested that HEV 239 antigen particle has better immunogenicity as well as antigenicity than those of NE2 antigen, so it is a better vaccine candidate against HEV.
Keywords:hepatitis E virus  capsid protein  vaccine  particulate antigen  prokaryotic ex pression
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