Mapping of a conformational epitope shared between E1 and E2 on the serum-derived human hepatitis C virus envelope
Mapping of a conformational epitope shared between E1 and E2 on the serum-derived human hepatitis C virus envelope
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DOI：
10.1074/jbc.m304047200
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发表时间：
2003-11-07
影响因子：
4.8
通讯作者：
Trépo, C
中科院分区：
文献类型：
--
作者：
Petit, MA;Jolivet-Reynaud, C;Trépo, C
通过用从慢性HCV 1b感染患者的血浆置换获得的富含丙型肝炎病毒（HCV）的沉淀免疫小鼠产生的单克隆抗体D32.10结合来自不同HCV 1a和HCV 1b感染患者的血清的HCV颗粒。此外，该单克隆抗体已显示识别HCV包膜蛋白E1和E2。为了提供对HCV包膜糖蛋白E1和E2的膜拓扑结构的新见解，我们使用Ph.D.- 12（TM）噬菌体展示肽文库技术。D32.10筛选的模拟表位与HCV E1糖蛋白的（297）RHWTTQGCNC（306）和HCV E2糖蛋白的（YRLWHYPCT 621）-Y-613和（480）PDQRPYCWHYPPKPC（494）具有部分相似性。D32.10与对应于这三个HCV区域的重叠肽的免疫反应性证实了这些定位，并表明所鉴定的三个区域可能在血清衍生颗粒的表面上紧密并列，如由来自蜱传脑炎病毒E蛋白的HCV E2的二级模型结构所预测的。这一论断得到了支持的检测针对这三个E1 E2区的HCV感染患者血清中的特异性抗体。
Monoclonal antibody D32.10 produced by immunizing mice with a hepatitis C virus (HCV)-enriched pellet obtained from plasmapheresis of a chronically HCV1b-infected patient binds HCV particles derived from serum of different HCV1a- and HCV1b-infected patients. Moreover, this monoclonal has been shown to recognize both HCV envelope proteins E1 and E2. In an attempt to provide novel insight into the membrane topology of HCV envelope glycoproteins E1 and E2, we localized the epitope recognized by D32.10 on the E1 and/or E2 sequence using Ph.D.-12(TM) phage display peptide library technology. Mimotopes selected from the phage display dodecapeptide library by D32.10 shared partial similarities with (297)RHWTTQGCNC(306) of the HCV E1 glycoprotein and with both (YRLWHYPCT621)-Y-613 and (480)PDQRPYCWHYPPKPC(494) of the HCV E2 glycoprotein. Immunoreactivity of D32.10 with overlapping peptides corresponding to these three HCV regions confirmed these localizations and suggested that the three regions identified are likely closely juxtaposed on the surface of serum-derived particles as predicted by the secondary model structure of HCV E2 derived from the tick-borne encephalitis virus E protein. This assertion was supported by the detection of specific antibodies directed against these three E1E2 regions in sera from HCV-infected patients.