生物法合成伤寒O-糖蛋白结合疫苗及其免疫原性评估

doi:10.16288/j.yczz.15-001

遗传 ›› 2015, Vol. 37 ›› Issue (5): 473-479.doi: 10.16288/j.yczz.15-001

生物法合成伤寒O-糖蛋白结合疫苗及其免疫原性评估

彭哲慧^1,2,潘超²,孙鹏²,冯尔玲²,吴军²,朱力²,彭清忠¹,王恒樑²

1. 吉首大学生物资源与环境科学学院,湖南吉首 416000;
2. 军事医学科学院生物工程研究所,病原微生物生物安全国家重点实验室,北京 100071

收稿日期:2015-01-25 出版日期:2015-05-20 发布日期:2015-05-20
通讯作者: 彭清忠,教授,研究方向：微生物制药。E-mail: qzpengjsu@163.com;王恒樑,研究员,研究方向：病原微生物。E-mail: wanghl@bmi.ac.cn
作者简介:彭哲慧,硕士研究生,专业方向：病原微生物。E-mail: michel121@126.com
基金资助:
国家自然科学基金项目(编号：81373316)资助

Preparation and immunogenicity-evaluation of typhoid O-specific polysaccharides bio-conjugate vaccines

Zhehui Peng^{1, 2}, Chao Pan², Peng Sun², Erling Feng², Jun Wu², Li Zhu², Qingzhong Peng¹, Hengliang Wang²

1. College of Biology and Environmental Sciences, Jishou University, Jishou 416000, China;
2. State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing 100071, China

Received:2015-01-25 Online:2015-05-20 Published:2015-05-20

摘要/Abstract

摘要： 伤寒由伤寒沙门氏菌(Salmonella Typhi)引发,至今在发展中国家仍是备受关注的重要公共卫生问题。文章通过敲除伤寒菌脂多糖合成途径中O-抗原连接酶基因,转入含脑膜炎奈瑟球菌(Neisseria meningitidis)蛋白糖基化途径中糖基转移酶的表达载体,以及改构的重组铜绿假单胞菌(Pseudomonas Aeruginosa)外毒素A(rEPA_N29)的表达载体,使细胞内能够诱导合成以伤寒O特异性多糖(O-specific polysaccharides, OPS)为目标抗原、以rEPA_N29为载体蛋白的伤寒OPS-rEPA_N29糖蛋白复合物,并对纯化所得复合物进行了免疫原性评价。ELISA测定血清抗体滴度表明,rEPA_N29作为载体蛋白能有效增加糖链的免疫原性,糖蛋白比单独的多糖能诱导产生更好的免疫应答;3次免疫、间隔3周比间隔2周IgG滴度稍有提高;而免疫过量的糖蛋白,抗O-多糖的血清抗体效价并无提升。文章为生物法制备多糖-蛋白结合疫苗提供了新思路,理论上也适用于其他革兰氏阴性菌的疫苗研发。

关键词: 结合疫苗, 伤寒沙门氏菌, 合成生物学, 重组EpA, O-多糖抗原

Abstract: Typhoid fever caused by Salmonella Typhi is still a major public health problem in developing countries. In this study, we constructed a genetically modified Salmonella Typhi strain expressing O-specific polysaccharides (OPS) antigen conjugated to a carrier, recombinant Pseudomonas aeruginosa exotoxin A(rEPA_N29). The conjugates (OPS-rEPA_N29) were further purified and evaluated for their immunogenicity. The results of ELISA showed that the conjugates evoked higher titers of IgG than OPS, suggesting that rEPA_N29 increased immunogenicity of OPS significantly as a carrier. Moreover, three injections with 3-week interval evoked slightly higher titers of IgG than three injections with 2-week interval. However, injection of excess conjugates could not evoke higher titers of IgG against lipid polysaccharide (LPS). In summary, our study provides a new strategy for preparing polysaccharides-protein conjugate vaccines as well as similar bio-conjugate vaccines of other Gram-negative pathogens.

Key words: conjugate vaccine, Salmonella Typhi, synthetic biology, rEPA, O-polysaccharide antigen

彭哲慧,潘超,孙鹏,冯尔玲,吴军,朱力,彭清忠,王恒樑. 生物法合成伤寒O-糖蛋白结合疫苗及其免疫原性评估[J]. 遗传, 2015, 37(5): 473-479.

Zhehui Peng, Chao Pan, Peng Sun, Erling Feng, Jun Wu, Li Zhu, Qingzhong Peng, Hengliang Wang. Preparation and immunogenicity-evaluation of typhoid O-specific polysaccharides bio-conjugate vaccines[J]. HEREDITAS(Beijing), 2015, 37(5): 473-479.

参考文献

[1] 2013年卫生统计提要, 卫生部网站: www.moh.gov.cn/ ewebeditor/uploadfile/2014/04/20140430131845405.pdf.
[2] Bhutta ZA. Impact of age and drug resistance on mortality in typhoid fever. Arch Dis Child , 1996, 75(3): 214-217.
[3] Lerouge I, Vanderleyden J. O-antigen structural variation: mechanisms and possible roles in animal/plant-microbe interactions. FEMS Microbio Rev , 2002, 26(1): 17-47.
[4] Faridmoayer A, Fentabil MA, Mills DC, Klassen JS, Feldman MF. Functional characterization of bacterial oligosaccharyltransferases involved in O-linked protein glycosylation. J Bacteriol , 2007, 189(22): 8088-8098.
[5] Hug I, Feldman MF. Analogies and homologies in lipopolysaccharide and glycoprotein biosynthesis in bacteria. Glycobiology , 2011, 21(2): 138-151.
[6] 王恒樑, 冯尔玲, 史兆兴, 姚潇, 苏国富, 黄留玉. 用Red系统快速敲除痢疾杆菌asd基因. 军事医学科学院院刊, 2002, 26(3): 172-175.
[7] Acosta CJ, Yang HH, Ning W, Qion G, Qun D, Ma XL, Zhou BD, Liu W, Carolina Danovaro-Holliday M, Leon Ochiai R, Wang XY, Kim DR, Xu ZY, Dong BQ, Galindo CM, Clemens JD. Efficacy of a locally produced, Chinese Vi polysaccharide typhoid fever vaccine during six years of follow-up. Vaccine , 2005, 23(48-49): 5618-5623.
[8] Szu SC, Stone AL, Robbins JD, Schneerson R, Robbins JB. Vi capsular polysaccharide-protein conjugates for prevention of typhoid fever. Preparation, characterization, and immunogenicity in laboratory animals. J Exper Med , 1987, 166(5): 1510-1524.
[9] Jones C. Vaccines based on the cell surface carbohydrates of pathogenic bacteria. An Acad Bras Cienc , 2005, 77(2): 293-324.
[10] Terra VS, Mills DC, Yates LE, Abouelhadid S, Cuccui J, Wren BW. Recent developments in bacterial protein glycan coupling technology and glycoconjugate vaccine design. J Med Microbiol , 2012, 61(Pt7): 919-926.
[11] Dagan R, Poolman J, Siegrist CA. Glycoconjugate vaccines and immune interference: A review. Vaccine , 2010, 28(34): 5513-5523.
[12] Passwell JH, Harlev E, Ashkenazi S, Chu C, Miron D, Ramon R, Farzan N, Shiloach J, Bryla DA, Majadly F, Roberson R, Robbins JB, Schneerson R. Safety and immunogenicity of improved Shigella O-specific polysaccharide-protein conjugate vaccines in adults in Israel. Infect Immun , 2001, 69(3): 1351-1357.
[13] Ahmed A, Li JP, Shiloach Y, Robbins JB, Szu SC. Safety and immunogenicity of Escherichia coli O157 O-specific polysaccharide conjugate vaccine in 2-5-year-old children. J Infect Dis , 2006, 193(4): 515-521.
[14] Ihssen J, Kowarik M, Dilettoso S, Tanner C, Wacker M, Thöny-Meyer L. Production of glycoprotein vaccines in Escherichia coli . Microb Cell Fac , 2010, 61: 1-13.
[15] Fernandez S, Palmer DR, Simmons M, Sun PF, Bisbing J, McClain S, Mani S, Burgess T, Gunther V, Sun W. Potential role for toll-like receptor 4 in mediating Escherichia coli maltose-binding protein activation of dendritic cells. Infect Immun , 2007, 75(3): 1359-1363.
[16] Paoletti LC, Kasper DL, Michon F, DiFabio J, Jennings HJ, Tosteson TD, Wessels MR. Effects of chain length on the immunogenicity in rabbits of group B streptococcus type III oligosaccharide-tetanus toxoid conjugates. J Clin Invest , 1992, 89(1): 203-209.

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生物法合成伤寒O-糖蛋白结合疫苗及其免疫原性评估

Preparation and immunogenicity-evaluation of typhoid O-specific polysaccharides bio-conjugate vaccines

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