Department of Virology and Parasitology, Fujita Health University School of Medicine, Toyoake, Aichi, Japan.
J Med Virol. 2014 Apr;86(4):558-67. doi: 10.1002/jmv.23734. Epub 2013 Sep 11.
In order to identify the repertoire of antibodies generated on natural infection of norovirus (NoV) in humans, and to characterize the human monoclonal antibodies against NoV, three phage-displayed antibody libraries originating from healthy person(s) were screened using purified virus-like particles (VLPs) of strain Narita 104 (r104, genogroup II, genotype 4) or strain Chiba 407 (rCV, genogroup I, genotype 4) as antigens. On screening with r104, 62 clones were isolated. Among these antibodies, two clones, 12A11 and 12B10, showed intra-genogroup cross-reactivity to genotypes 1, 3-7, 12, and 14, and genotypes 1, 4, 6, and 7 of genogroup II, respectively. In addition, antibodies belonging to the same group were isolated from two different libraries. On screening with rCV, five clones were isolated, two of which were cross-reactive. One, CV-2F5, reacted to genotypes 1-4, and 8 of genogroup I, and the other, CV-1A5, showed inter-genogroup cross-reactivity to all the VLPs employed in this study. The blocking activities of the monoclonal antibodies against the interaction of homotypic VLPs (VLPs used in the panning procedure) with histo-blood group antigens were also assessed as an alternative to neutralization assay. Although the blocking activity of 12A11 was partially limited 12B10 prevented the binding of r104 to histo-blood group antigens that had been reported to bind r104. The blocking activity of CV-2F5 against the attachment of rCV to suitable histo-blood group antigens was weak, but the blocking activity of CV-1A5 was well recognized. Thus, 12B10 and CV-1A5 were suggested to be cross-reactive monoclonal antibodies with neutralizing activity.
为了鉴定人类感染诺如病毒（NoV）时产生的抗体库，并鉴定针对 NoV 的人源单克隆抗体，我们使用纯化的病毒样颗粒（VLPs）Narita 104 株（r104，基因型 II，基因型 4）或 Chiba 407 株（rCV，基因型 I，基因型 4）作为抗原，筛选了三个源自健康个体的噬菌体展示抗体文库。在用 r104 筛选时，分离出 62 个克隆。在这些抗体中，两个克隆 12A11 和 12B10 对基因型 I、3-7、12 和 14 以及基因型 II 的 1、4 显示了同种型交叉反应性，分别为 1、4、6 和 7。此外，来自两个不同文库的同种型抗体被分离出来。在用 rCV 筛选时，分离出 5 个克隆，其中 2 个具有交叉反应性。一个是 CV-2F5，它与基因型 I 的 1-4 和 8 反应，另一个是 CV-1A5，它与本研究中使用的所有 VLPs 表现出种间交叉反应性。还评估了单克隆抗体对同种型 VLPs（筛选过程中使用的 VLPs）与组织血型抗原相互作用的阻断活性，作为中和测定的替代方法。尽管 12A11 的阻断活性部分受到限制，但 12B10 阻止了已报道与 r104 结合的组织血型抗原与 r104 的结合。CV-2F5 对 rCV 与合适的组织血型抗原结合的阻断活性较弱，但 CV-1A5 的阻断活性得到了很好的识别。因此，12B10 和 CV-1A5 被认为是具有中和活性的交叉反应性单克隆抗体。
