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猪圆环病毒衣壳蛋白抗原表位的精细定位及2型猪圆环病毒的抗原表型

Fine mapping of antigenic epitopes on capsid proteins of porcine circovirus, and antigenic phenotype of porcine circovirus type 2.

作者信息

Shang Shao-Bin, Jin Yu-Lan, Jiang Xue-tao, Zhou Ji-Yong, Zhang Xin, Xing Gang, He Jia Ling, Yan Yan

机构信息

Key Laboratory of Animal Epidemic Etiology & Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, PR China.

出版信息

Mol Immunol. 2009 Jan;46(3):327-34. doi: 10.1016/j.molimm.2008.10.028. Epub 2008 Dec 6.

Abstract

Type 2 porcine circovirus (PCV2) is associated with post-weaning multisystemic wasting syndrome in pigs. In this study, three monoclonal antibodies (mAbs) against the capsid protein (Cap) of PCV2, eight mAbs to Cap of type 1 porcine circovirus (PCV1) and five mAbs specific for Cap of both PCV1 and PCV2, were generated and used to finely map the antigenic sites of PCV1 and PCV2, and to identify the antigenic phenotype of PCV2 with different length of genome. Five linear B-cell epitopes, including the residues 231-233 and 195-202 specific for PCV2, residues 92-103 specific for PCV1, and residues 156-162 and 175-192 shared between PCV1 and PCV2, were finely defined with synthetic peptides, and the critical residue in epitope 231-233 and 156-162 was located at residues 233 ((233)Proline) and 156 ((156)Tyrosine), respectively. The conformational epitopes recognized by mAbs with neutralizing activity against both PCV1 and PCV2 were detected in transfected PK-15 and the residues 231-233 also participated in the formation of conformational epitopes. Analysis of antigenic diversity on these epitopes exhibited three antigenic phenotypes of PCV2, (1766)PCV2, (1767)PCV2 and (1768)PCV2 using mAbs. The results from this study first demonstrated the different antigenic phenotype between PCV2 isolates.

摘要

2型猪圆环病毒(PCV2)与猪断奶后多系统消耗综合征有关。在本研究中,制备了三种针对PCV2衣壳蛋白(Cap)的单克隆抗体(mAb)、八种针对1型猪圆环病毒(PCV1)Cap的mAb以及五种对PCV1和PCV2的Cap均特异的mAb,并用于精细定位PCV1和PCV2的抗原位点,以及鉴定具有不同基因组长度的PCV2的抗原表型。利用合成肽精细确定了五个线性B细胞表位,包括PCV2特异的231 - 233位和195 - 202位残基、PCV1特异的92 - 103位残基以及PCV1和PCV2共有的156 - 162位和175 - 192位残基,表位231 - 233和156 - 162中的关键残基分别位于233位((233)脯氨酸)和156位((156)酪氨酸)。在转染的PK - 15细胞中检测到了对PCV1和PCV2均具有中和活性的mAb所识别的构象表位,231 - 233位残基也参与了构象表位的形成。利用mAb对这些表位的抗原多样性分析显示PCV2有三种抗原表型,即(1766)PCV2、(1767)PCV2和(1768)PCV2。本研究结果首次证明了PCV2分离株之间存在不同的抗原表型。

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