Unit of Vaccinology and Antiviral Strategies, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Viruses. 2018 Jan 5;10(1):27. doi: 10.3390/v10010027.
Noroviruses (NoVs) are the main pathogens responsible for sporadic and epidemic nonbacterial gastroenteritis, causing an estimated 219,000 deaths annually worldwide. There is no commercially available vaccine for NoVs, due partly to the difficulty in establishing NoV cell culture models. The histo-blood group antigen (HBGA) blocking assay is used extensively to assess the protective potential of candidate vaccine-elicited antibodies, but there is still no widely used cellular evaluation model. In this study, we have established a cell line-based NoV vaccine evaluation model through the construction of human α1,2-fucosyltransferase 2-overexpressing 293T (293T-FUT2) cell lines. The 293T-FUT2 cells stably expressed H type 2 and Lewis y antigens. Virus-like particles (VLPs) of the NoV prototype strain genogroup I.1 (GI.1) and the predominant strains GII.4 and GII.17 could attach to the cell line efficiently in a dose-dependent manner. Importantly, antisera against these NoV VLPs could inhibit the attachment of the VLPs, where the inhibitory effects measured by the attachment inhibition assay correlated significantly with the antibody levels determined by the HBGA blocking assay. Collectively, our attachment inhibition assay could serve as a surrogate neutralization assay for the evaluation of NoV vaccines at the cellular level.
诺如病毒（NoV）是导致散发性和流行性非细菌性胃肠炎的主要病原体，每年在全球造成约 21.9 万人死亡。由于难以建立诺如病毒细胞培养模型，目前尚无市售的诺如病毒疫苗。组织血型抗原（HBGA）阻断试验广泛用于评估候选疫苗诱导抗体的保护潜力，但仍没有广泛使用的细胞评估模型。在这项研究中，我们通过构建人α1,2-岩藻糖基转移酶 2 过表达 293T（293T-FUT2）细胞系，建立了基于细胞系的诺如病毒疫苗评估模型。293T-FUT2 细胞稳定表达 H 型 2 和 Lewis y 抗原。诺如病毒原型株基因组 I.1（GI.1）和主要株 GII.4 和 GII.17 的病毒样颗粒（VLPs）能够以剂量依赖的方式有效地附着在细胞系上。重要的是，针对这些诺如病毒 VLPs 的抗血清能够抑制 VLPs 的附着，其中通过附着抑制试验测量的抑制作用与通过 HBGA 阻断试验确定的抗体水平显著相关。总之，我们的附着抑制试验可以作为细胞水平评估诺如病毒疫苗的替代中和试验。
