Comprehensive Mapping Antigenic Epitopes of NS1 Protein of Japanese Encephalitis Virus with Monoclonal Antibodies.
Comprehensive Mapping Antigenic Epitopes of NS1 Protein of Japanese Encephalitis Virus with Monoclonal Antibodies.
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DOI：
10.1371/journal.pone.0067553
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Bu ZG
中科院分区：
文献类型：
--
作者：
Hua RH;Liu LK;Chen ZS;Li YN;Bu ZG
日本脑炎病毒（JEV）非结构蛋白1（NS1）参与病毒复制并在感染过程中产生保护性免疫应答。JEV NS1特异性抗体反应可作为鉴别诊断不同黄病毒感染的指标。然而，JEV NS1上的表位的特征很差。本研究描述了完整的定位线性B细胞表位在JEV NS1。我们从用重组NS1免疫的小鼠中产生了11个NS1特异性单克隆抗体。对于单克隆抗体的表位作图，用GST标签表达一组覆盖整个NS1蛋白的51个部分重叠的肽，然后使用单克隆抗体进行筛选。通过酶联免疫吸附试验（ELISA），确定了五个线性含表位的肽。通过从肽的羧基和氨基末端依次去除氨基酸残基，使用单克隆抗体鉴定并确认了五个线性表位的最小单元。五个线性表位位于氨基酸残基5AIDITRK11、72RDELNVL78、251KSKHNRREGY 260、269DENGIVLD 276和341DETTLVRS 348中。通过序列比对发现，JEV毒株间的表位高度保守。值得注意的是，当测试交叉反应性时，来自其他黄病毒的NS1蛋白上的同源区域都不与MAb反应，并且所有五个表位肽都不被抗西尼罗河病毒或登革病毒的血清识别。这些新的JEV NS1病毒特异性线性B细胞表位将有利于开发新的疫苗和诊断方法。
Japanese encephalitis virus (JEV) non-structural protein 1 (NS1) contributes to virus replication and elicits protective immune responses during infection. JEV NS1-specific antibody responses could be a target in the differential diagnosis of different flavivirus infections. However, the epitopes on JEV NS1 are poorly characterized. The present study describes the full mapping of linear B-cell epitopes in JEV NS1. We generated eleven NS1-specific monoclonal antibodies from mice immunized with recombinant NS1. For epitope mapping of monoclonal antibodies, a set of 51 partially-overlapping peptides covering the entire NS1 protein were expressed with a GST-tag and then screened using monoclonal antibodies. Through enzyme-linked immunosorbent assay (ELISA), five linear epitope-containing peptides were identified. By sequentially removing amino acid residues from the carboxy and amino terminal of peptides, the minimal units of the five linear epitopes were identified and confirmed using monoclonal antibodies. Five linear epitopes are located in amino acids residues 5AIDITRK11, 72RDELNVL78, 251KSKHNRREGY260, 269DENGIVLD276, and 341DETTLVRS348. Furthermore, it was found that the epitopes are highly conserved among JEV strains through sequence alignment. Notably, none of the homologous regions on NS1 proteins from other flaviviruses reacted with the MAbs when they were tested for cross-reactivity, and all five epitope peptides were not recognized by sera against West Nile virus or Dengue virus. These novel virus-specific linear B-cell epitopes of JEV NS1 would benefit the development of new vaccines and diagnostic assays.
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