Identification of B-cell epitopes in the NSP1 protein of porcine reproductive and respiratory syndrome virus
Identification of B-cell epitopes in the NSP1 protein of porcine reproductive and respiratory syndrome virus
复制标题
猪繁殖与呼吸综合征病毒NSP1蛋白B细胞表位的鉴定
DOI：
10.1016/j.vetmic.2011.09.018
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发表时间：
2012-03-23
影响因子：
3.3
通讯作者：
Jiang, Ping
中科院分区：
文献类型：
--
作者：
Song, Yanhua;Zhou, Yefei;Jiang, Ping
猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus，PRRSV）分为北美型和欧洲型。NSP 1是PRRSV重要的非结构蛋白，由复制酶多聚蛋白切割成NSP 1 α和NSP 1 β亚基，在免疫抑制中起重要作用。本研究通过在大肠杆菌中表达猪繁殖与呼吸综合征病毒（PRRSV）NSP 1基因，筛选并鉴定了6株抗该基因的单克隆抗体。Western blot和IFA结果表明，6株单克隆抗体中有4株识别重组NSP 1 α，2株识别重组NSP 1 β。单克隆抗体4 H2识别NSP 1 α的线性表位E（54）EPLRW（59），单克隆抗体2G 5、3E 11和4D 4识别NSP 1 α的线性表位H（157）VLTNLP（163），单克隆抗体3C 7和1H 7识别NSP 1 β的线性表位185 ~ 232 aa。NSP 1蛋白质序列比对表明，E（54）EPLRW（59）在所有北美PRRSV毒株中是保守的，而欧洲型毒株在该区域具有可变的氨基酸。表位H（157）VLTNLP（163）在所有的PRRSV毒株中相对保守，除了欧洲型毒株的L162 -> S162改变。表位185- 232 aa在北美PRRSV毒株中是可变的。这些结果为进一步研究PRRSV NSP 1的功能和PRRSV感染的诊断方法奠定了基础。(C)2011 Elsevier B. V.保留所有权利。
Porcine reproductive and respiratory syndrome virus (PRRSV) was divided into North American and European genotypes. NSP1 was an important non-structural protein of PRRSV, which was auto-cleaved from the replicase polyprotein into NSP1 alpha and NSP1 beta subunits and played an important role in the immune suppression. In this study, six monoclonal antibodies (MAbs) against the recombinant PRRSV NSP1, expressed in Escherichia coli system, were screened out and identified. Western blot and IFA results indicated that 4 out of 6 MAbs recognized the recombinant NSP1 alpha and 2 MAbs recognized NSP1 beta. Epitope mapping results indicated that MAb 4H2 recognized the linear epitopes E(54)EPLRW(59) in NSP1 alpha, MAbs (2G5, 3E11 and 4D4) recognized the epitopes H(157)VLTNLP(163) in NSP1 alpha, and MAbs 3C7 and 1H7 reacted with the epitopes 185aa to 232aa in NSP1 beta. Protein sequence alignment of NSP1 indicated E(54)EPLRW(59) was conserved in all North American PRRSV strains, whereas European type strains has variable amino acids in this region. The epitope H(157)VLTNLP(163) was relatively conserved among all PRRSV strains, except for a L162 -> S162 change in European type strains. The epitope 185-232aa was variable among North American PRRSV strains. These results may facilitate future investigations into the function of NSP1 of PRRSV and diagnostic methods for PRRSV infection. (C) 2011 Elsevier B.V. All rights reserved.