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Identification of H-2d Restricted T Cell Epitope of Foot-and-mouth Disease Virus Structural Protein VP1

机译:口蹄疫病毒结构蛋白VP1的H-2d限制性T细胞表位的鉴定

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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.
机译:背景技术口蹄疫(FMD)是一种高度传染性和毁灭性的疾病,影响牲畜,造成重大的经济损失。因此,需要针对口蹄疫病毒(FMDV)的更安全,更有效的疫苗。这项研究的目的是从FMD存在的病毒结构蛋白VP1中筛选并鉴定H-2d限制性T细胞表位。因此,我们提供了研究特定FMDV T细胞表位的方法和基础。结果采用密码子优化表达方法有效表达VP1蛋白在结肠杆菌中。我们使用口蹄疫标准阳性血清用于其免疫原性的Western印迹检测。 VP1蛋白用于免疫BALB / c小鼠,并分离了脾淋巴细胞。然后,对通过使用生物信息学方法预测和合成的VP1蛋白上潜在的H-2Dd,H-2Kd和H-2Ld限制性T细胞表位进行了常见的体外训练刺激。使用淋巴细胞增殖测定法和IFN-γELISPOT实验鉴定了H-2Kd限制性T细胞抗原决定簇pK1(AYHKGPFTRL)和H-2Dd限制性T细胞抗原决定簇pD7(GFIMDRFVKI)。结论这项研究的结果为研究FMDV免疫过程,疫苗开发等奠定了基础。这些结果还表明,鉴定病毒T细胞表位,生物信息学和分子生物学方法的联合应用是有效的。

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