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猪T细胞针对口蹄疫病毒结构蛋白的抗原特异性:VP1中T辅助表位的鉴定

Antigenic specificity of porcine T cell response against foot-and-mouth disease virus structural proteins: identification of T helper epitopes in VP1.

作者信息

Rodríguez A, Sáiz J C, Novella I S, Andreu D, Sobrino F

机构信息

CISA-INIA, Valdeolmos, Madrid, Spain.

出版信息

Virology. 1994 Nov 15;205(1):24-33. doi: 10.1006/viro.1994.1616.

Abstract

The contribution of each of the viral capsid proteins of foot-and-mouth disease virus (FMDV) in the T cell response of vaccinated pigs has been studied. Viral polypeptides, VP1 to VP4, were expressed as fusion proteins in Escherichia coli, and were used to stimulate peripheral blood mononuclear cells of vaccinated animals. Significant, dose-dependent responses to whole virion were detected in the seven animals analyzed and, in five of them, responses to recombinant polypeptides VP1, VP2, and VP3 were noticed, VP4 was recognized only by one of the pigs. Among the responder animals, VP1 and VP3 induced the higher proliferative responses. The patterns of recognition of a nested set of VP3 fragments expressed as fusions in E. coli were different among the animals studied and were consistent with the presence of different T cell epitopes on the protein. Likewise, three of the four VP1 fragments induced significant responses and were differentially recognized by each of the animals tested. Partially overlapping synthetic peptides spanning VP1 amino acids 41 to 209 were used to identify T cell epitopes in this protein. The significant responses obtained in three of seven additional FMDV vaccinated outbred pigs analyzed revealed the existence of at least 11 different T cell epitopes distributed throughout the sequence studied, which were distinctly recognized by each of the responder animals. A peptide corresponding to a relevant B cell antigenic site, around amino acids 140-160, was shown to stimulate lymphocytes from two of the responder animals. Thus, the results obtained indicate that different T cell epitopes of capsid proteins VP1 and VP3 are recognized by pig populations. The different patterns of recognition of recombinant polypeptides and synthetic peptides observed among outbred animals support an important contribution of genetic restriction, probably mediated by MHC genes, to the individual T cell response in swine.

摘要

对口蹄疫病毒(FMDV)的每种病毒衣壳蛋白在接种疫苗猪的T细胞反应中的作用进行了研究。病毒多肽VP1至VP4在大肠杆菌中作为融合蛋白表达,并用于刺激接种动物的外周血单核细胞。在所分析的7只动物中检测到对完整病毒粒子有显著的剂量依赖性反应,其中5只动物对重组多肽VP1、VP2和VP3有反应,只有1只猪识别出VP4。在有反应的动物中,VP1和VP3诱导了更高的增殖反应。在研究的动物中,以融合形式在大肠杆菌中表达的一组嵌套VP3片段的识别模式不同,这与该蛋白上存在不同的T细胞表位一致。同样,四个VP1片段中的三个诱导了显著反应,并且被每个测试动物以不同方式识别。使用跨越VP1氨基酸41至209的部分重叠合成肽来鉴定该蛋白中的T细胞表位。在另外7只接种FMDV的远交猪中,有3只获得了显著反应,这表明在所研究的序列中存在至少11种不同的T细胞表位,每个有反应的动物都能明显识别这些表位。一个对应于大约氨基酸140 - 160处相关B细胞抗原位点的肽被证明能刺激两只反应动物的淋巴细胞。因此,获得的结果表明衣壳蛋白VP1和VP3的不同T细胞表位被猪群体识别。在远交动物中观察到的重组多肽和合成肽的不同识别模式支持了遗传限制(可能由MHC基因介导)对猪个体T细胞反应的重要作用。

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