Inhibition of Hepatitis C Virus Infection by Anti-Claudin-1 Antibodies Is Mediated by Neutralization of E2-CD81-Claudin-1 Associations
Inhibition of Hepatitis C Virus Infection by Anti-Claudin-1 Antibodies Is Mediated by Neutralization of E2-CD81-Claudin-1 Associations
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DOI：
10.1002/hep.23445
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发表时间：
2010-04-01
期刊：
影响因子：
13.5
通讯作者：
Baumert, Thomas F.
中科院分区：
文献类型：
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作者：
Krieger, Sophie E.;Zeisel, Mirjam B.;Baumert, Thomas F.
紧密连接蛋白claudin-1（CLDN 1）已被证明是丙型肝炎病毒（HCV）进入病毒感染的第一步所必需的。由于缺乏中和抗CLDN 1抗体，CLDN 1在病毒进入过程中的作用知之甚少。在这项研究中，我们通过基因免疫产生了针对人CLDN 1细胞外环的抗体，并使用这些抗体来研究CLDN 1在感染性HCV细胞培养系统和人肝细胞中对HCV进入的机制作用。对细胞表面表达的CLDN 1具有特异性的抗体以剂量依赖性方式特异性抑制HCV感染。与单独使用的任一种试剂相比，对CLDN 1、清道夫受体B1和CD 81具有特异性的抗体显示出相加的中和能力。抗CLDN 1和抗CD 81抗体的动力学研究表明，HCV与两种进入因子的相互作用发生在内化过程中的相似时间。在不存在可检测的CLDN 1-E2相互作用的情况下，抗CLDN 1抗体抑制包膜糖蛋白E2与HCV容许细胞系的结合。使用荧光标记的进入因子和荧光共振能量转移方法，我们证明了抗CLDN 1抗体抑制CD 81-CLDN 1缔合。相比之下，CLDN 1-CLDN 1和CD 81-CD 81关联不受调节。总之，我们的结果表明，靶向CLDN 1的抗体通过减少E2与细胞表面的结合并破坏CD 81-CLDN 1相互作用来中和HCV感染性。结论：这些结果进一步确定了CLDN 1在HCV进入过程中的功能，并突出了靶向E2-CD 81-CLDN 1相互作用的新的抗病毒策略。（《肝脏学》2010年;51：1144-1157）
The tight junction protein claudin-1 (CLDN1) has been shown to be essential for hepatitis C virus (HCV) entry-the first step of viral infection. Due to the lack of neutralizing anti-CLDN1 antibodies, the role of CLDN1 in the viral entry process is poorly understood. In this study, we produced antibodies directed against the human CLDN1 extracellular loops by genetic immunization and used these antibodies to investigate the mechanistic role of CLDN1 for HCV entry in an infectious HCV cell culture system and human hepatocytes. Antibodies specific for cell surface expressed CLDN1 specifically inhibit HCV infection in a dose-dependent manner. Antibodies specific for CLDN1, scavenger receptor B1, and CD81 show an additive neutralizing capacity compared with either agent used alone. Kinetic studies with anti-CLDN1 and anti-CD81 antibodies demonstrate that HCV interactions with both entry factors occur at a similar time in the internalization process. Anti-CLDN1 antibodies inhibit the binding of envelope glycoprotein E2 to HCV permissive cell lines in the absence of detectable CLDN1-E2 interaction. Using fluorescent-labeled entry factors and fluorescence resonance energy transfer methodology, we demonstrate that anti-CLDN1 antibodies inhibit CD81-CLDN1 association. In contrast, CLDN1-CLDN1 and CD81-CD81 associations were not modulated. Taken together, our results demonstrate that antibodies targeting CLDN1 neutralize HCV infectivity by reducing E2 association with the cell surface and disrupting CD81-CLDN1 interactions. Conclusion: These results further define the function of CLDN1 in the HCV entry process and highlight new antiviral strategies targeting E2-CD81-CLDN1 interactions. (HEPATOLOGY 2010;51:1144-1157.)