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