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鉴定口蹄疫病毒结构蛋白 VP1 中的 H-2d 限制性 T 细胞表位。

Identification of H-2d restricted T cell epitope of foot-and-mouth disease virus structural protein VP1.

机构信息

State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.

出版信息

Virol J. 2011 Sep 7;8:426. doi: 10.1186/1743-422X-8-426.

Abstract

BACKGROUND

Foot-and-mouth disease (FMD) is a highly contagious and devastating disease affecting livestock that causes significant financial losses. Therefore, safer and more effective vaccines are required against Foot-and-mouth disease virus(FMDV). The purpose of this study is to screen and identify an H-2d restricted T cell epitope from the virus structural protein VP1, which is present with FMD. We therefore provide a method and basis for studying a specific FMDV T cell epitope.

RESULTS

A codon-optimized expression method was adopted for effective expression of VP1 protein in colon bacillus. We used foot-and-mouth disease standard positive serum was used for Western blot detection of its immunogenicity. The VP1 protein was used for immunizing BALB/c mice, and spleen lymphocytes were isolated. Then, a common in vitro training stimulus was conducted for potential H-2Dd, H-2Kd and H-2Ld restricted T cell epitope on VP1 proteins that were predicted and synthesized by using a bioinformatics method. The H-2Kd restricted T cell epitope pK1 (AYHKGPFTRL) and the H-2Dd restricted T cell epitope pD7 (GFIMDRFVKI) were identified using lymphocyte proliferation assays and IFN-γ ELISPOT experiments.

CONCLUSIONS

The results of this study lay foundation for studying the FMDV immune process, vaccine development, among other things. These results also showed that, to identify viral T cell epitopes, the combined application of bioinformatics and molecular biology methods is effective.

摘要

背景

口蹄疫(FMD)是一种高度传染性和破坏性的家畜疾病,会导致重大经济损失。因此,需要针对口蹄疫病毒(FMDV)开发更安全、更有效的疫苗。本研究旨在从病毒结构蛋白 VP1 中筛选和鉴定与 FMD 相关的 H-2d 限制性 T 细胞表位,为研究特定的 FMDV T 细胞表位提供方法和依据。

结果

采用密码子优化表达方法,在大肠杆菌中有效表达 VP1 蛋白。采用口蹄疫标准阳性血清进行 Western blot 检测,证明其具有免疫原性。用 VP1 蛋白免疫 BALB/c 小鼠,分离脾淋巴细胞。然后,采用生物信息学方法预测和合成 VP1 蛋白上可能存在的 H-2Dd、H-2Kd 和 H-2Ld 限制性 T 细胞表位,并进行体外共同刺激。通过淋巴细胞增殖试验和 IFN-γ ELISPOT 实验,鉴定出 H-2Kd 限制性 T 细胞表位 pK1(AYHKGPFTRL)和 H-2Dd 限制性 T 细胞表位 pD7(GFIMDRFVKI)。

结论

本研究结果为研究 FMDV 免疫过程、疫苗开发等奠定了基础。结果还表明,通过生物信息学和分子生物学方法的联合应用,可以有效地鉴定病毒 T 细胞表位。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5518/3179754/b00a8aa42535/1743-422X-8-426-1.jpg

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