细菌性腹泻三联口服疫苗的研制及其免疫效果的初步评价

doi:10.13523/j.cb.20170706

中国生物工程杂志

2017, Vol. 37

Issue (7): 18-26 DOI: 10.13523/j.cb.20170706

研究报告

细菌性腹泻三联口服疫苗的研制及其免疫效果的初步评价

刘地¹, 晏婷¹, 何秀娟², 郑文云², 马兴元¹

1. 华东理工大学生物工程学院生物反应器工程国家重点实验室上海 200237;
2. 华东理工大学药学院上海市新药设计重点实验室上海 200237

Preparation and Preliminary Evaluation of Triple Oral Vaccine Against Bacterial Diarrhea

LIU Di¹, YAN Ting¹, HE Xiu-juan², ZHENG Wen-yun², MA Xing-yuan¹

1. School of Biotechnology and State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;
2. School of Pharmacy, Shanghai Key Laboratory of New Drug Design, East China University of Science and Technology, Shanghai 200237, China

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摘要： 由细菌引起的感染性腹泻，至今仍是世界范围广泛流行的传染病之一。由于耐抗生素病原菌的不断涌现，导致了抗生素药物的治疗效果不佳。因此，研发便捷、有效的疫苗对于细菌性腹泻的预防与治疗尤为重要。针对产肠毒素大肠杆菌、霍乱弧菌与志贺氏痢疾菌这三种最为主要的细菌性腹泻病原菌，设计和筛选了以热不稳定肠毒素亚基蛋白为抗原和黏膜佐剂、霍乱弧菌鞭毛蛋白及志贺毒素亚基蛋白为抗原的三联疫苗。通过工程大肠杆菌获得了相应的抗原与佐剂蛋白，并以海藻酸钙-壳聚糖微球为疫苗的口服载体，制备了疫苗的口服制剂。体外实验表明，微球载体中的蛋白质在模拟胃液中释放较低，但在模拟肠液中释放迅速，这种载体能够实现疫苗在肠道内定向释放的目的。通过对灌胃免疫后小鼠免疫指标的检测，证明了疫苗能够刺激机体产生抗原特异性的sIgA与IgG抗体，免疫组与对照组相比，差异显著（P<0.05），并提升了外周血中CD4⁺T细胞的含量（7.5%~9.5%）与CD4⁺T/CD8⁺T细胞的比率，有效地激活了机体的黏膜免疫与系统免疫，能够对机体起到保护作用。

关键词： 细菌性腹泻; 黏膜免疫; 三联口服疫苗

Abstract: Infectious diarrhea disease caused by bacterial pathogens is still one of the most common infectious diseases throughout the world. Due to the emergence of antibiotic resistant bacteria, the treatment effects of antibiotic drugs are not very well. Therefore, it is very important to develop convenient and effective vaccines for the prevention and treatment of bacterial diarrhea. Aimed at enterotoxigenic Escherichia coli (ETEC), Vibrio cholerae and Shigella flexneri, the three major types of pathogenic bacteria, the triple-vaccine based on the heat-labile enterotoxin subunit proteins as antigens and mucosal adjuvants, Vibrio cholerae flagellin protein and Shiga toxin subunit protein as antigens was designed. The corresponding antigens and adjuvant proteins were obtained from engineered Escherichia coli, the alginate-chitosan microspheres were used as oral delivery, and the oral vaccine formulations were prepared. In vitro experiment indicate that the release of protein is very slow in simulated gastric fluid, but prompt and almost complete in simulated intestinal fluid, which suggests that microsphere carrier can achieve the purpose of the intestinal targeted release of vaccine. After immunizing mice, immune indexes were detected, and the results proved that the vaccine stimulated the production of the antigen-specific sIgA and IgG antibody, with significant difference (P<0.05) compared with the control group, and improved the content of CD4⁺T cells (7.5%~9.5%) and the ratio of CD4⁺T/CD8⁺T cells in peripheral blood. As a result, the vaccine induced mucosal and systemic immune responses and could effectively protect the body after immunization.

Key words: Bacterial diarrhea Mucosal immunity Triple oral vaccine

收稿日期: 2017-04-21 出版日期: 2017-07-25

ZTFLH:

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通讯作者: 马兴元 E-mail: xymy@ecust.edu.cn

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引用本文:

刘地, 晏婷, 何秀娟, 郑文云, 马兴元. 细菌性腹泻三联口服疫苗的研制及其免疫效果的初步评价[J]. 中国生物工程杂志, 2017, 37(7): 18-26.

LIU Di, YAN Ting, HE Xiu-juan, ZHENG Wen-yun, MA Xing-yuan. Preparation and Preliminary Evaluation of Triple Oral Vaccine Against Bacterial Diarrhea. China Biotechnology, 2017, 37(7): 18-26.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170706 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I7/18

[1] Sanchez J,Holmgren J. Virulence factors, pathogenesis and vaccine protection in cholera and ETEC diarrhea. Current Opinion in Immunology, 2005, 17(4): 388-398.
[2] Liu L, Johnson H L, Cousens S, et al. Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet, 2012, 379(9832): 2151-2161.
[3] Zaidi D,Wine E. An update on travelers’ diarrhea. Curr Opin Gastroenterol, 2015, 31(1): 7-13.
[4] Das J K, Tripathi A, Ali A, et al. Vaccines for the prevention of diarrhea due to cholera, shigella, ETEC and rotavirus. BMC Public Health, 2013, 13(Suppl 3): S11.
[5] Wenneras C,Erling V. Prevalence of enterotoxigenic Escherichia coli-associated diarrhoea and carrier state in the developing world. J Health Popul Nutr, 2004, 22(4): 370-382.
[6] Svennerholm A M,Steele D. Microbial-gut interactions in health and disease. Progress in enteric vaccine development. Best Pract Res Clin Gastroenterol, 2004, 18(2): 421-445.
[7] Peirano G, Souza F S,Rodrigues D P. Frequency of serovars and antimicrobial resistance in Shigella spp. from Brazil. Memorias do Instituto Oswaldo Cruz, 2006, 101(3): 245-250.
[8] Das J K, Ali A, Salam R A, et al. Antibiotics for the treatment of Cholera, Shigella and Cryptosporidium in children. BMC Public Health, 2013, 13(Suppl 3): S10.
[9] Ogra P L, Faden H,Welliver R C. Vaccination strategies for mucosal immune responses. Clinical Microbiology Reviews, 2001, 14(2): 430-445.
[10] Hoebe K, Janssen E,Beutler B. The interface between innate and adaptive immunity. Nature Immunology, 2004, 5(10): 971-974.
[11] da Hora V P, Conceicao F R, Dellagostin O A, et al. Non-toxic derivatives of LT as potent adjuvants. Vaccine, 2011, 29(8): 1538-1544.
[12] Rappuoli R, Pizza M, Douce G, et al. Structure and mucosal adjuvanticity of cholera and Escherichia coli heat-labile enterotoxins. Immunology Today, 1999, 20(11): 493-500.
[13] Melton-Celsa A R. Shiga toxin (Stx) classification, structure, and function. Microbiol Spectr, 2014, 2(4): Ehec-0024-2013.
[14] Klose K E,Mekalanos J J. Differential regulation of multiple flagellins in Vibrio cholerae. Journal of Bacteriology, 1998, 180(2): 303-316.

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