Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2009, Vol. 29 Issue (03): 20-24    
研究报告     
抗禽传染性支气管炎病毒多肽疫苗EpiA免疫原性研究
阳泰1,王红宁1,汪雪1,2,高荣1,唐俊妮1,李玉玲1,郭自城1
1. 四川大学生命科学学院
2. 四川农业大学动物医学院
The Immune Response Against Avian Infectious Bronchitis Virus Induced by Multi-epitope based Peptide Vaccines
 全文: PDF(598 KB)   HTML
摘要:

设计基于B细胞表位和T细胞表位的禽传染性支气管炎病毒(IBV)多肽疫苗EpiA,并利用基因工程重组技术将EpiA在大肠杆菌中表达、纯化。ELISA和Western blot法验证其免疫原性后,将重组蛋白配以弗氏佐剂免疫鸡,并以灭活苗、GST和PBS为试验对照组。细胞免疫效应采用流式细胞仪(FACS)对免疫鸡外周血中CD4+ ,CD8+ T淋巴细胞进行计数,IBV特异性抗体采用ELISA法进行检测,并进行攻毒试验。结果显示,成功设计了多肽疫苗EpiA:ELISA和Western blot 证明所表达的融合蛋白能与标准IBV阳性血清产生特异性抗原-抗体反应,而且该融合蛋白能有效激发鸡体特异性体液和细胞免疫,与对照组差异显著(P≤0.01)。攻毒试验表明,多肽疫苗保护率可达73%,大肠杆菌生物合成重组抗IBV多肽疫苗将为IBV疫苗研究制备提供了新的途径。

关键词: IBV;多肽疫苗;设计;免疫原性    
Abstract:

In this study, peptide vaccine was designed based on B cell and T cell epitope of IBV, and the multi-epitope based peptide vaccine was expressed in E.coli fused with glutathione S-transferase (GST). ELISA and Western blot analysis showed that purified fusion proteins (EpiA) had an excellent immune activity with chicken anti-IBV serum. The expressed fusion proteins were purified and emulsified with Freund adjuvant. Vaccination twice at an interval of 2 weeks with the emulsified the fusion proteins elicited anti-IBV specific antibodies and specific cellular responses and 73% of the vaccinated chickens were protected against mortality. We proposed use of a biosynthesis system instead of peptide synthesis, provided a new way to prepare multi-epitope based peptide vaccine

Key words: IBV;Peptide vaccine;Design;immunogenic
收稿日期: 2008-11-05 出版日期: 2009-03-31
基金资助:

2006AA10A205;国家“863”计划资助项目

通讯作者: 王红宁   
服务  
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章  
阳泰 王红宁 汪雪 高荣 唐俊妮 李玉玲 郭自城

引用本文:

阳泰,王红宁,汪雪,高荣,唐俊妮,李玉玲,郭自城. 抗禽传染性支气管炎病毒多肽疫苗EpiA免疫原性研究[J]. 中国生物工程杂志, 2009, 29(03): 20-24.

YANG Tai- Wang-Gong-Ning- Hong-Xue- Gao-Rong- Tang-Dun-Ni- Li-Yu-Ling- Guo-Zi-Cheng. The Immune Response Against Avian Infectious Bronchitis Virus Induced by Multi-epitope based Peptide Vaccines. China Biotechnology, 2009, 29(03): 20-24.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2009/V29/I03/20

[1] Avellaneda G E, Villeges P, Jackwood M W, et al. In vivo evaluation of the pathogenecity of isolates of infectious bronchitis virus. Avian Diseases, 1994, 38:589~597 [2] 孙立枢,唐宏琨,陈五岭. 两种树脂对固相法合成丙型肝炎疫苗多肽的影响. 中国生物工程杂志,2007,27(2):80~84 Sun L S, Tang H K, Cheng W L. China Biotechnology, 2007, 27 (2):80~84 [3] Purcell A W, McCluskey J,Rossjoh J. More than one reason to rethink the use of peptides in vaccine design. Nat Rev Drug Discov, 2007, 6:404~414 [4] Lenstra J A, Kusters J G, Koch G, et al.Antigenicity of the peplomer protein of infectious bronchitis virus. Mol Immunol, 1989, 26:7~15 [5] Wang L, Parr R L, King D J, et al.A highly conserved epitope on the spike protein of infectious bronchitis virus. Arch Virol, 1995,140:2201~2213 [6] Boots A M, Kusters J G, Van Noort J M, et al.Localization of a Tcell epitope within the nucleocapsid protein of avian coronavirus. Immunology,1991,74:8~13 [7] Partidos C,Stanley C,Steward M. The influence of orientation and number of copies of T and B cell epitopes on the specificity and affinity of antibodies induced by chimeric peptide. European Journal of Immunology ,1992, 22: 2675~2680 [8] Holzhutter H G, Frommel C,Kloetzel P M. A theoretical approach towards the identification of cleavagedetermining amino acid motifs of the 20 S proteasome. J Mol Biol, 1999, 286:1251~1265 [9] Li G J, Chen W Z, Yan W Y, et al. Comparison of immune responses against footandmouth disease virus induced by fusion proteins using the swine IgG heavy chain constant region or hgalactosidase as a carrier of immunogenic epitopes. Virology, 2004, 328, 274~281

No related articles found!