Monoclonal Antibodies against Extracellular Domains of Claudin-1 Block Hepatitis C Virus Infection in a Mouse Model
Monoclonal Antibodies against Extracellular Domains of Claudin-1 Block Hepatitis C Virus Infection in a Mouse Model
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DOI：
10.1128/jvi.03676-14
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发表时间：
2015-05-01
影响因子：
5.4
通讯作者：
Kondoh, Masuo
中科院分区：
文献类型：
--
作者：
Fukasawa, Masayoshi;Nagase, Shotaro;Kondoh, Masuo
丙型肝炎病毒 (HCV) 进入宿主细胞是一个复杂的过程，需要多种宿主因子，包括claudin-1 (CLDN1)。需要开发安全有效的治疗性进入抑制剂。我们分离了一种人类肝Huh7.5.1衍生的细胞突变体，该突变体对HCV不敏感，并且比较微阵列分析表明该突变体具有CLDN1缺陷。获得了四个杂交瘤，它们产生的单克隆抗体 (MAb) 与亲本 Huh7.5.1 细胞相互作用，但不与 CLDN1 缺陷突变体相互作用。这些杂交瘤产生的所有单克隆抗体均以非常高的亲和力与人 CLDN1 特异性结合，并以剂量依赖性方式阻止 HCV 对 Huh7.5.1 细胞的感染，且没有明显的细胞毒性。两种选定的 MAb 还可以抑制人肝嵌合小鼠的 HCV 感染，且没有明显的副作用。 CLDN1可能是体内预防HCV感染的潜在靶点。因此，抗 CLDN1 MAb 可能成为新型抗 HCV 药物的有希望的候选者。 重要性 针对丙型肝炎病毒 (HCV) 的安全有效的治疗性进入抑制剂对于与其他抗 HCV 药物（例如直接作用抗病毒药物）的联合治疗非常有用。在这项研究中，我们首先展示了一种有效的策略，通过使用表达完整靶膜蛋白的亲代细胞和靶缺陷细胞，开发针对多膜靶蛋白 Claudin-1 (CLDN1) 胞外域的功能性单克隆抗体 (MAb)。所建立的针对 CLDN1 的单克隆抗体对完整的 CLDN1 具有非常高的亲和力，可有效抑制体外和体内 HCV 感染。这些抗 CLDN1 单克隆抗体有望成为新型 HCV 进入抑制剂的先导物。
Hepatitis C virus (HCV) entry into host cells is a complex process requiring multiple host factors, including claudin-1 (CLDN1). Safe and effective therapeutic entry inhibitors need to be developed. We isolated a human hepatic Huh7.5.1-derived cell mutant that is nonpermissive to HCV, and comparative microarray analysis showed that the mutant was CLDN1 defective. Four hybridomas were obtained, which produced monoclonal antibodies (MAbs) that interacted with the parental Huh7.5.1 cell but not with the CLDN1-defective mutant. All MAbs produced by these hybridomas specifically bound to human CLDN1 with a very high affinity and prevented HCV infection of Huh7.5.1 cells in a dose-dependent manner, without apparent cytotoxicity. Two selected MAbs also inhibited HCV infection of human liver-chimeric mice without significant adverse effects. CLDN1 may be a potential target to prevent HCV infection in vivo. Anti-CLDN1 MAbs may hence be promising candidates as novel anti-HCV agents.IMPORTANCESafe and effective therapeutic entry inhibitors against hepatitis C virus (HCV) are very useful for combination therapies with other anti-HCV drugs, such as direct-acting antivirals. In this study, we first showed an effective strategy for developing functional monoclonal antibodies (MAbs) against extracellular domains of a multimembrane-spanning target protein, claudin-1 (CLDN1), by using parental cells expressing the intact target membrane protein and target-defective cells. The established MAbs against CLDN1, which had a very high affinity for intact CLDN1, efficiently inhibited in vitro and in vivo HCV infections. These anti-CLDN1 MAbs are promising leads for novel entry inhibitors against HCV.