重组酿酒酵母表达幽门螺杆菌VacA蛋白及其免疫原性分析<sup>*</sup>

doi:10.13523/j.cb.1912044

中国生物工程杂志

2020, Vol. 40

Issue (5): 15-21 DOI: 10.13523/j.cb.1912044

研究报告

重组酿酒酵母表达幽门螺杆菌VacA蛋白及其免疫原性分析^*

岑黔鸿,高彤,任怡,雷涵(

)

西南交通大学医学院成都 610031

Recombinant Saccharomyces cerevisiae Expressing Helicobacter pylori VacA Protein and Its Immunogenicity Analysis

CEN Qian-hong,GAO Tong,REN Yi,LEI Han(

)

College of Medicine, Southwest Jiaotong University, Chengdu 610031, China

全文: PDF(943 KB) HTML

摘要：

目的:利用酿酒酵母表面展示技术筛选幽门螺杆菌候选疫苗,并分析其免疫原性。方法:以幽门螺杆菌的空泡型细胞毒素A(vacA)基因作为研究对象,构建重组S.cerevisiae EBY100/pYD1-VacA,通过Western blot、免疫荧光标记和流式细胞仪对S.cerevisiae EBY100/pYD1-VacA进行体外表达分析。以PBS和S.cerevisiae EBY100/pYD1为对照组,S.cerevisiae EBY100/pYD1-VacA为实验组,口服免疫SPF级BALB/c小鼠。通过ELISA分析检测口服免疫后小鼠抗VacA特异性IgG及分泌型IgA效价。结果:VacA抗原蛋白被成功地展示在S.cerevisiae EBY100表面。小鼠经口服免疫S.cerevisiae EBY100/pYD1-VacA后可诱导产生较高的VacA特异性抗体。结论:表面展示型酿酒酵母可以作为幽门螺杆菌候选疫苗的递送载体,与此同时,这也为开发其他细菌或病毒疫苗提供新思路。

关键词： 酿酒酵母表面展示技术; S.cerevisiae EBY100/pYD1-VacA; 口服免疫; 免疫原性

Abstract:

Objective: Helicobacter pylori candidate vaccine was constructed based on yeast surface display technology and its immunogenicity was further analyzed. Methods: Surface displayed S.cerevisiae EBY100/pYD1-VacA was constructed that vacA gene of Helicobacter pylori as used a research subject. S.cerevisiae EBY100/pYD1-VacA was detected by Western blot analysis,immunofluorescence assay and flow cytometry assay. BALB/c mice of SPF grade were administrated orally with that PBS and S.cerevisiae EBY100/pYD1 were used as the control groups, and S.cerevisiae EBY100/pYD1-VacA was used as the experimental group. VacA-specific sera IgG and secretory IgA titers were determined by ELISA assay. Results: The VacA antigen protein was successfully displayed on the surface of S.cerevisiae EBY100. S.cerevisiae EBY100/pYD1-VacA could produce a higher VacA specific antibodies. Conclusion: Surface displayed yeast can be used as a delivery vector of Helicobacter pylori candidate vaccine. Meanwhile, it will provide new ideas for developing bacteria or virus vaccines.

Key words: Saccharomyces cerevisiae surface display technology S.cerevisiae EBY100/pYD1-VacA Oral immunization Immunogenicity

收稿日期: 2019-12-24 出版日期: 2020-06-02

ZTFLH:

Q815

基金资助: * 四川省科技计划重点面上项目(2019YFN0134);中央高校基本科研业务费专项资金(2682018CX73)

通讯作者: 雷涵 E-mail: hlei@swjtu.edu.cn

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引用本文:

岑黔鸿,高彤,任怡,雷涵. 重组酿酒酵母表达幽门螺杆菌VacA蛋白及其免疫原性分析^*[J]. 中国生物工程杂志, 2020, 40(5): 15-21.

CEN Qian-hong,GAO Tong,REN Yi,LEI Han. Recombinant Saccharomyces cerevisiae Expressing Helicobacter pylori VacA Protein and Its Immunogenicity Analysis. China Biotechnology, 2020, 40(5): 15-21.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.1912044 或 https://manu60.magtech.com.cn/biotech/CN/Y2020/V40/I5/15

图1 vacA基因的PCR结果

图2 重组表达质粒pYD1-VacA的酶切鉴定电泳图

图3 S.cerevisiae EBY100/pYD1-VacA的PCR鉴定电泳图

图4 Western blot分析

图5 免疫荧光分析

图6 流式细胞仪分析

图7 VacA特异性抗体应答

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