Isolation of human monoclonal antibodies to the envelope e2 protein of hepatitis C virus and their characterization.
Isolation of human monoclonal antibodies to the envelope e2 protein of hepatitis C virus and their characterization.
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DOI：
10.1371/journal.pone.0055874
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Hotta H
中科院分区：
文献类型：
--
作者：
Shimizu YK;Hijikata M;Oshima M;Shimizu K;Alter HJ;Purcell RH;Yoshikura H;Hotta H
我们分离并鉴定了两种抗丙型肝炎病毒（HCV）包膜E2蛋白的人单克隆抗体。稳定产生抗体的淋巴母细胞系通过使用EB病毒使慢性丙型肝炎患者的外周血单核细胞永生化而获得。用从同一患者分离的表达HCV株H（基因型1a）的E2蛋白的Huh 7细胞，通过免疫荧光进行抗体阳性克隆的筛选。得到的抗体#37和#55的同种型分别是IgG 1/κ和IgG 1/λ。表位作图显示#37和#55分别识别跨越氨基酸429至652和508至607的构象表位。通过使用病毒感染的Huh7.5细胞作为靶标的免疫荧光，两种抗体与测试的所有九种不同的HCV基因型/亚型反应。抗体显示出不同的免疫染色模式;而#37产生主要位于细胞核周边的颗粒状反应，#55在整个细胞质中产生弥漫性染色。免疫金电子显微镜显示两种抗体均与完整的病毒颗粒结合。在中和试验中（使用含有源自基因型1a、1b、2a、2b、3a、4a、5a、6a和7a的结构蛋白的嵌合感染性HCV进行病灶形成单位减少），#55抑制除基因型7a以外的所有测试HCV基因型的感染的程度较低。#37没有中和任何这些病毒。作为广泛交叉中和的人抗体，#55可用于HCV感染的被动免疫治疗。
We isolated and characterized two human monoclonal antibodies to the envelope E2 protein of hepatitis C virus (HCV). Lymphoblastoid cell lines stably producing antibodies were obtained by immortalizing peripheral blood mononuclear cells of a patient with chronic hepatitis C using Epstein-Barr virus. Screening for antibody-positive clones was carried out by immunofluorescence with Huh7 cells expressing the E2 protein of HCV strain H (genotype 1a) isolated from the same patient. Isotype of resulting antibodies, #37 and #55, was IgG1/kappa and IgG1/lambda, respectively. Epitope mapping revealed that #37 and #55 recognize conformational epitopes spanning amino acids 429 to 652 and 508 to 607, respectively. By immunofluorescence using virus-infected Huh7.5 cells as targets both antibodies were reactive with all of the nine different HCV genotypes/subtypes tested. The antibodies showed a different pattern of immuno-staining; while #37 gave granular reactions mostly located in the periphery of the nucleus, #55 gave diffuse staining throughout the cytoplasm. Both antibodies were shown by immuno-gold electron microscopy to bind to intact viral particles. In a neutralization assay (focus-forming unit reduction using chimeric infectious HCV containing structural proteins derived from genotypes 1a, 1b, 2a, 2b, 3a, 4a, 5a, 6a, and 7a), #55 inhibited the infection of all HCV genotypes tested but genotype 7a to a lesser extent. #37 did not neutralize any of these viruses. As a broadly cross-neutralizing human antibody, #55 may be useful for passive immunotherapy of HCV infection.
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DOI：
10.1073/pnas.0504877103
发表时间：
2006-05-09
影响因子：
11.1
作者：
Pietschmann, Thomas;Kaul, Artur;Bartenschlager, Ralf
通讯作者：
Bartenschlager, Ralf
影响因子：
2.5
作者：
Shimizu, Y. K.;Hijikata, M.;Yoshikura, H.
通讯作者：
Yoshikura, H.
影响因子：
5.4
作者：
Perotti, Mario;Mancini, Nicasio;Burioni, Roberto
通讯作者：
Burioni, Roberto
影响因子：
5.4
作者：
Blight, KJ;McKeating, JA;Rice, CM
通讯作者：
Rice, CM
影响因子：
29.4
作者：
Gottwein, Judith M.;Scheel, Troels K. H.;Bukh, Jens
通讯作者：
Bukh, Jens