Inserm, U748, Strasbourg, France.
Gastroenterology. 2010 Sep;139(3):953-64, 964.e1-4. doi: 10.1053/j.gastro.2010.05.073. Epub 2010 Jun 2.
BACKGROUND & AIMS: Hepatitis C virus (HCV) infection is a challenge to prevent and treat because of the rapid development of drug resistance and escape. Viral entry is required for initiation, spread, and maintenance of infection, making it an attractive target for antiviral strategies. The tight junction protein claudin-1 (CLDN1) has been shown to be required for entry of HCV into the cell.
Using genetic immunization, we produced 6 monoclonal antibodies against the host entry factor CLDN1. The effects of antibodies on HCV infection were analyzed in human cell lines and primary human hepatocytes.
Competition and binding studies demonstrated that antibodies interacted with conformational epitopes of the first extracellular loop of CLDN1; binding of these antibodies required the motif W(30)-GLW(51)-C(54)-C(64) and residues in the N-terminal third of CLDN1. The monoclonal antibodies against CLDN1 efficiently inhibited infection by HCV of all major genotypes as well as highly variable HCV quasispecies isolated from individual patients. Furthermore, antibodies efficiently blocked cell entry of highly infectious escape variants of HCV that were resistant to neutralizing antibodies.
Monoclonal antibodies against the HCV entry factor CLDN1 might be used to prevent HCV infection, such as after liver transplantation, and might also restrain virus spread in chronically infected patients.
由于耐药性和逃逸的快速发展,丙型肝炎病毒(HCV)感染的防治极具挑战性。病毒进入是病毒起始、传播和维持感染所必需的,因此成为抗病毒策略的一个有吸引力的靶点。紧密连接蛋白 Claudin-1(CLDN1)已被证明是 HCV 进入细胞所必需的。
我们通过遗传免疫产生了针对宿主进入因子 CLDN1 的 6 种单克隆抗体。在人细胞系和原代人肝细胞中分析了抗体对 HCV 感染的影响。
竞争和结合研究表明,抗体与 CLDN1 的第一个细胞外环的构象表位相互作用;这些抗体的结合需要 motif W(30)-GLW(51)-C(54)-C(64)和 CLDN1 的 N 端三分之一的残基。针对 CLDN1 的单克隆抗体可有效抑制所有主要基因型 HCV 以及从个体患者中分离出的高度可变 HCV 准种的感染。此外,抗体还能有效阻止对中和抗体具有耐药性的高度感染性逃逸变异 HCV 的细胞进入。
针对 HCV 进入因子 CLDN1 的单克隆抗体可用于预防 HCV 感染,例如肝移植后,也可限制慢性感染患者的病毒传播。