基于酿酒酵母表面展示技术构建寨卡疫苗及其免疫活性初探

基于酿酒酵母表面展示技术构建寨卡疫苗及其免疫活性初探

黄子恩,张炜,岑黔鸿,耿耘,雷涵

西南交通大学生命科学与工程学院,西南交通大学医学院,西南交通大学医学院,西南交通大学生命科学与工程学院;西南交通大学医学院,西南交通大学医学院

收稿日期:2018-04-17 修回日期:2018-05-03 出版日期:2018-04-25
基金资助:
中央高校基本科研业务费专项资金资助(项目编号：2682016YXZT06)

Construction of ZIKV Oral Vaccine Based on Saccharomyces cerevisiae Surface Display Technology and the Preliminary Study on Its Immune Activity

HUANG Zien,ZHANG Wei,CENG Qianhong,GENG Yun and LEI Han

South West JiaoTong University School of Life Science and Engineering,Sichuan,South West JiaoTong University College of Medicine,Sichuan,South West JiaoTong University College of Medicine,Sichuan,South West JiaoTong University School of Life Science and Engineering,Sichuan;South West JiaoTong University College of Medicine,Sichuan,South West JiaoTong University School of Life Science and Engineering,Sichuan;South West JiaoTong University College of Medicine,Sichuan

Received:2018-04-17 Revised:2018-05-03 Online:2018-04-25

摘要/Abstract

摘要： 目的：基于酿酒酵母表面展示技术构建具有免疫活性的寨卡病毒（ZIKV）口服疫苗。方法：以Zika/SZ01/2016的包膜（Envelope）基因作为研究对象，以酿酒酵母表面展示质粒pYD1为骨架，构建EBY100/pYD1-Envelope。通过蛋白质印迹法（western blot）、免疫荧光分析和流式细胞仪分析，对Envelope蛋白进行定性分析，通过BCA蛋白测定试剂盒对Envelope蛋白进行定量分析。以SPF级的BALB/c小鼠作为动物模型，考察3种免疫方案（给药方案A：50 OD600 nm·d-1×3 d，给药方案B：75 OD600 nm·d-1×2 d，给药方案C：150 OD600 nm·d-1×1 d）的口服免疫效果。结果： 3种免疫方案均能诱发产生较高水平的体液免疫应答和黏膜免疫应答。给药方案C的免疫效果最佳。结论：本研究首次探讨了ZIKV口服疫苗的免疫效果，为有效预防ZIKV感染提供了一种可供参考的解决方案。

Abstract: Objective： To construct Zika virus（ZIKV） oral vaccine based on the Saccharomyces cerevisiae (S.cerevisiae) surface display technology. Methods： EBY100/pYD1-Envelope was constructed based on the Envelope gene of Zika/SZ01/2016 used as research subject and S.cerevisiae surface display plasmid pYD1 was used for a backbone. Qualitative analysis was performed by western blot， immunofluorescence assay and flow cytometric assay； quantitative analysis was then made by BCA protein detecting kit as well. SPF grade BALB/c mice were used as animal model to study the oral immune effect of 3 dosage regimens (Regimen A：50 OD600 nm·d-1×3 d，Regimen B：75 OD600 nm·d-1×2 d，Regimen C：150 OD600 nm·d-1×1 d). Results： All the three regimens could induce higher levels of humoral immune response and mucosal response. Regimen C was preferred. Conclusion： In this study， it is the first time to evaluate the immune efficacy of ZIKV oral vaccine. Moreover， it provides a reference strategy for effective prevention of ZIKV infection.

黄子恩,张炜,岑黔鸿,耿耘,雷涵. 基于酿酒酵母表面展示技术构建寨卡疫苗及其免疫活性初探[J]. .

HUANG Zien,ZHANG Wei,CENG Qianhong,GENG Yun and LEI Han. Construction of ZIKV Oral Vaccine Based on Saccharomyces cerevisiae Surface Display Technology and the Preliminary Study on Its Immune Activity[J]. .

参考文献

[1] Larocca R A, Abbink P, Peron J P, et al. Vaccine protection against Zika virus from Brazil [J]. Nature, 2016, 536(7617): 474-478.
[2] Dowd K A, Ko S Y, Morabito K M, et al. Rapid development of a DNA vaccine for Zika virus [J]. Science, 2016, 354(6309): 237-240.
[3] Abbink P, Larocca R A, De La Barrera R A, et al. Protective efficacy of multiple vaccine platforms against Zika virus challenge in rhesus monkeys [J]. Science, 2016, 353(6304): 1129-1132.
[4] Muthumani K, Griffin B D, Agarwal S, et al. In vivo protection against ZIKV infection and pathogenesis through passive antibody transfer and active immunisation with a prMEnv DNA vaccine [J]. npj Vaccines, 2016, 1(1): 16021.
[5] Pardi N, Hogan M J, Pelc R S, et al. Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination [J]. Nature, 2017, 543(7644): 248-251.
[6] Richner J M, Himansu S, Dowd K A, et al. Modified mRNA vaccines protect against Zika virus infection [J]. Cell, 2017, 168(6): 1114-1125.
[7] Shan Chao, Muruato A E, Nunes B T D, et al. A live-attenuated Zika virus vaccine candidate induces sterilizing immunity in mouse models [J]. Nat Med, 2017, 23(6): 763-767.
[8] Kim E, Erdos G, Huang S, et al. Preventative vaccines for Zika virus outbreak: preliminary evaluation [J]. EBioMedicine, 2016, 13: 315-320.
[9] Li Xiao-feng, Dong Hao-long, Wang Hong-jiang, Wang H J, et al. Development of a chimeric Zika vaccine using a licensed live-attenuated flavivirus vaccine as backbone [J]. Nat Commun, 2018, 9(1): 673.
[10] Dick G W, Kitchen S F, Haddow A J. Zika virus. I. Isolations and serological specificity [J]. Trans R Soc Trop Med Hyg, 1952, 46(5): 509-520.
[11] Duffy M R, Chen T H, Hancock W T, et al. Zika virus outbreak on Yap Island, Federated States of Micronesia [J]. N Engl J Med, 2009, 360(24): 2536-2543.
[12] Li Jian-dong, Xiong Ying, Wu Wei, et al. Zika Virus in a Traveler Returning to China from Caracas, Venezuela, February 2016 [J]. Emerg Infect Dis, 2016, 22(6): 1133-1136.
[13] Gyawali N, Bradbury R S, Taylor-Robinson A W. The global spread of Zika virus: is public and media concern justified in regions currently unaffected? [J]. Infect Dis Poverty, 2016, 5: 37.
[14] Schuler-Faccini L, Ribeiro E M, Feitosa I M, et al. Possible Association Between Zika Virus Infection and Microcephaly - Brazil, 2015 [J]. MMWR Morb Mortal Wkly Rep, 2016, 65(3): 59-62.
[15] Parra B, Lizarazo J, Jimenez-Arango J A, et al. Guillain-Barre Syndrome Associated with Zika Virus Infection in Colombia [J]. N Engl J Med, 2016, 375(16): 1513-1523.
[16] Czerkinsky C, Holmgren J. Topical immunization strategies [J]. Mucosal Immunol, 2010, 3(6): 545-555.
[17] Akada R. Genetically modified industrial yeast ready for application [J]. J Biosci Bioeng, 2002, 94(6): 536-544.
[18] Sun Hui, Wang Long-jiang, Wang Tian-tian, et al. Display of Eimeria tenella EtMic2 protein on the surface of Saccharomyces cerevisiae as a potential oral vaccine against chicken coccidiosis [J]. Vaccine, 2014, 32(16): 1869-1876.
[19] Han Thomas, Sui Jian-hua, Bennett A S, et al. Fine epitope mapping of monoclonal antibodies against hemagglutinin of a highly pathogenic H5N1 influenza virus using yeast surface display [J]. Biochem Biophys Res Commun, 2011, 409(2): 253-259.
[20] Lei Han, Jin Sha, Karlsson E, et al. Yeast Surface-Displayed H5N1 Avian Influenza Vaccines [J]. J Immunol Res, 2016, 2016: 4131324.
[21] Stubbs A C, Martin K S, Coeshott C, et al. Whole recombinant yeast vaccine activates dendritic cells and elicits protective cell-mediated immunity [J]. Nat Med, 2001, 7(5): 625-629.

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