研究报告Ⅱ型脊灰病毒抗原表位嵌合蛋白的免疫学研究

doi:10.13523/j.cb.20160801

中国生物工程杂志

2016, Vol. 36

Issue (8): 1-8 DOI: 10.13523/j.cb.20160801

研究报告

研究报告Ⅱ型脊灰病毒抗原表位嵌合蛋白的免疫学研究

王晶¹, 夏文跃¹, 潘小霞², 赵冰心¹, 滕玉梅¹, 李传印¹, 杨筱舟¹, 文喻玲¹, 陈元鼎¹

1. 中国医学科学研究院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室昆明 650118;
2. 云南民族大学化学与生物技术学院民族药资源化学国家民委-教育部重点实验室昆明 650118

Immunological Activity of a Chimeric Protein Carrying an Epitope of Type 2 Poliovirus on the VP6 of Rotavirus as a Vector

WANG Jing¹, XIA Wen-yue¹, PAN Xiao-xia², ZHAO Bing-xin¹, TENG Yu-mei¹, LI Chuan-yin¹, YANG Xiao-zhou¹, WEN Yu-ling¹, CHEN Yuan-ding¹

1. Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Diseases, Kunming 650118, China;
2. Key Laboratory of Chemistry in Ethnic Medicinal Resources, State Ethnic Affairs Commission and Ministry of Education, Yunnan Minzu University, Kunming 650500, China

全文: PDF(1302 KB) HTML

摘要：

目的：评价以轮状病毒（RV）重组VP6蛋白为载体插入Ⅱ型脊髓灰质炎病毒（PV2）VP1蛋白上的1个抗原表位构建而成的嵌合蛋白的体外免疫学性质。方法：采用分子克隆和基因重组技术将PV2抗原表位插入到RV载体蛋白上，在大肠杆菌中表达并用SDS-PAGE确认表达产物，再通过动物免疫、Western blot、免疫荧光和病毒血清抗体中和试验分析嵌合蛋白的免疫学性质。结果：成功构建了以VP6为载体的PV2抗原表位嵌合蛋白6F/PV₂N₁，并且在E.coli系统中高效表达，嵌合蛋白免疫的豚鼠血清抗体对RV和PV2具备较好的中和活性。结论：以RV VP6为载体构建的嵌合蛋白具有较好的免疫原性，免疫豚鼠产生血清抗体可中和RV和PV2在体外细胞上的感染；进一步为研发RV/PV2嵌合疫苗提供了较好的基础。

关键词： 轮状病毒; 抗原表位; 脊灰病毒; 嵌合蛋白; 免疫原性

Abstract:

Objective:To evaluate immunological activity of chimeric protein carrying one PV2 antigenic epitope on rotavirus VP6. Methods:one chimeric protein was constructed by inserting an epitope derived from neutralizing antigenic sites 1 of PV2 in rotavirus VP6 as a vector through gene cloning and recombination; the chimeric protein was expressed in E.coli cells and detected by Western blot,immunofluorescence assay and neutralization test. Results:The chimeric protein was favorably constructed and expressed in E.coli cells efficiently. Antibodies induced in inoculated guinea pigs by the chimeric protein could both neutralize the infection of RV and PV2 in vitro. Conclusion:The constructed chimeric protein possesses dual immunogenicity and can elicit production of antibodies in immunized guinea pigs which can protect infection of RV in MA104 cells and PV in Vero cells. The results are valuable for development of new RV/PV2 bivalent vaccine.

Key words: Poliovirus Chimeric protein Epitope Rotavirus Immunogenicity

收稿日期: 2016-05-23 出版日期: 2016-08-25

ZTFLH:

Q819

基金资助:

云南省自然科学基金资助项目（2013FZ130，2012FD039）

通讯作者: 陈元鼎 E-mail: chenyd@imbcams.com.cn

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引用本文:

王晶, 夏文跃, 潘小霞, 赵冰心, 滕玉梅, 李传印, 杨筱舟, 文喻玲, 陈元鼎. 研究报告Ⅱ型脊灰病毒抗原表位嵌合蛋白的免疫学研究[J]. 中国生物工程杂志, 2016, 36(8): 1-8.

WANG Jing, XIA Wen-yue, PAN Xiao-xia, ZHAO Bing-xin, TENG Yu-mei, LI Chuan-yin, YANG Xiao-zhou, WEN Yu-ling, CHEN Yuan-ding. Immunological Activity of a Chimeric Protein Carrying an Epitope of Type 2 Poliovirus on the VP6 of Rotavirus as a Vector. China Biotechnology, 2016, 36(8): 1-8.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160801 或 https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I8/1

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