| 表达猪瘟病毒E2蛋白的重组腺病毒的构建及其在兔体内的免疫原性分析 |
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| 引用本文： | 孙元,祁巧芬,梁冰冰,成丹,李娜,于墨洋,王宇飞,刘霓虹,朱庆虎,仇华吉.表达猪瘟病毒E2蛋白的重组腺病毒的构建及其在兔体内的免疫原性分析[J].生物工程学报,2008,24(10):1734-1739. |
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| 作者姓名： | 孙元 祁巧芬 梁冰冰 成丹 李娜 于墨洋 王宇飞 刘霓虹 朱庆虎 仇华吉 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室猪传染病研究室,哈尔滨,150001 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 为了构建猪瘟重组腺病毒载体疫苗,通过细菌内同源重组法构建了含有猪瘟病毒E2基因的重组腺病毒rAdV-E2.测定其一步生长曲线,同时用间接免疫荧光试验和Western blotting检测外源基因表达,然后用rAdV-E2免疫家兔,免疫后6周用猪瘟兔化弱毒疫苗株(c株)进行攻击,攻毒后3 d取其脾脏,用实时荧光定量RT-PCR检测C株病毒RNA.结果表明,该重组腺病毒传至第10代时,毒价可达1.0×1010TCID<,50/mL;外源基因可在其中得到稳定表达;rAdV-E2接种兔免疫后2周产生猪瘟特异性抗体,免疫后5 W抗体达到峰值,攻毒后rAdV-E2接种兔和C株接种兔均未出现定型热反应,从其脾脏也未检测到C株病毒RNA,而野生型腺病毒接种兔均出现了定型热反应,并且从其脾脏检测大量C株病毒RNA,其含量达到了103拷贝/μL以上.由此表明,rAdV-E2可望开发为猪瘟候选疫苗.
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| 关 键 词： | 猪瘟病毒 E2基因 重组腺病毒 免疫原性 家兔 |
| 收稿时间： | 2008/3/16 0:00:00 |
Generation and Immunogenicity of a Recombinant Adenovirus Expressing the E2 Protein of Classical Swine Fever Virus in Rabbits |
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| Yuan Sun,Qiaofen Qi,Bingbing Liang,Dan Cheng,Na Li,Moyang Yu,Yufei Wang,Nihong Liu,Qinghu Zhu and Huaji Qiu.Generation and Immunogenicity of a Recombinant Adenovirus Expressing the E2 Protein of Classical Swine Fever Virus in Rabbits[J].Chinese Journal of Biotechnology,2008,24(10):1734-1739. |
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| Authors: | Yuan Sun Qiaofen Qi Bingbing Liang Dan Cheng Na Li Moyang Yu Yufei Wang Nihong Liu Qinghu Zhu and Huaji Qiu |
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| Institution: | Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China;Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Harbin 15000, China |
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| Abstract: | Classical swine fever (CSF), which is caused by classical swine fever virus (CSFV), causes significant losses in pig industry in many countries in Asia and Europe. The E2 glycoprotein of CSFV is the main target for neutralizing antibodies. In this study, a recombinant replication-defective human adenovirus expressing the CSFV E2 gene (rAdV-E2) was generated and evaluated for the immunogenicity in rabbits. The results showed that the rabbits immunized with rAdV-E2 developed high-level CSFV-specific antibodies. The rAdV-E2-immunized rabbits were all free of the regular fever and the viral replication in the spleen upon challenge with C-strain, which were seen in the rabbits immunized with the parent adenovirus of rAdV-E2. This indicates that the recombinant adenovirus can be an attractive candidate vaccine against CSF. |
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| Keywords: | classical swine fever virus E2 gene recombinant adenovirus immunogenicity rabbit |
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