Xu Jialun, Zhan Xiangjun, Wang Fengqing, Lyu Chenang, An Ran, Hewitt Joanne, Tian Peng, Wang Dapeng
Department of Food Science and Technology, School of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Rd., Shanghai, 200240, China.
Institute of Environmental Science and Research (ESR Ltd), Porirua, Wellington, New Zealand.
Food Environ Virol. 2025 Sep 27;17(4):50. doi: 10.1007/s12560-025-09664-0.
Human norovirus (HuNoV) is the leading cause of acute nonbacterial gastroenteritis globally. Histo-blood group antigens (HBGAs) have been recognized as attachment factors for HuNoV. However, the receptors or other potential attachment factors for HuNoV have not been fully elucidated. A bacterial cell surface-displayed system was used to express GII.4 HuNoV protruding domain (GII.4 P) to capture attachment factors from a diarrheal sample of a patient infected with GII.4 HuNoV. A library of candidate HuNoV proteinaceous attachment factors was constructed. Through a database comparison, a literature review, the predicted protein subcellular localization, the protein-protein interactions (PPIs) network analysis, the molecular simulation and the protein docking analysis, four candidate attachment factors were selected for further investigation from the initial 118 candidates. Finally, two proteins, namely myelin-oligodendrocyte glycoprotein (MOG) and leucine-rich repeat-containing protein 15 (LRRC15), were demonstrated strongly binding to GII.4 P by both in vivo and in vitro assays. The binding between MOG/LRRC15 and GII.4 P can be blocked by GII.4 HuNoV antibody. Immunofluorescence showed that MOG/LRRC15 and GII.4 HuNoV co-localized in the cytoplasm in transfected cells. However, GII.4 HuNoV did not proliferate in the newly constructed MOG/LRRC15 overexpressing cells. This study describes attempts to identify HuNoV proteinaceous attachment factors from the diarrheal sample. Findings from this study will aid in understanding of HuNoV infection in humans, enrichment of GII.4 HuNoV from various environments and construction of cell lines for HuNoV cultivation in vitro.
人诺如病毒（HuNoV）是全球急性非细菌性胃肠炎的主要病因。组织血型抗原（HBGA）已被确认为HuNoV的附着因子。然而，HuNoV的受体或其他潜在附着因子尚未完全阐明。利用细菌细胞表面展示系统表达GII.4 HuNoV突出结构域（GII.4 P），以从感染GII.4 HuNoV的患者腹泻样本中捕获附着因子。构建了候选HuNoV蛋白质附着因子文库。通过数据库比较、文献综述、预测的蛋白质亚细胞定位、蛋白质-蛋白质相互作用（PPI）网络分析、分子模拟和蛋白质对接分析，从最初的118个候选因子中选择了4个候选附着因子进行进一步研究。最后，通过体内和体外试验证明，髓鞘少突胶质细胞糖蛋白（MOG）和富含亮氨酸重复序列的蛋白15（LRRC15）这两种蛋白与GII.4 P有强烈结合。MOG/LRRC15与GII.4 P之间的结合可被GII.4 HuNoV抗体阻断。免疫荧光显示，MOG/LRRC15与GII.4 HuNoV在转染细胞的细胞质中共定位。然而，GII.4 HuNoV在新构建的MOG/LRRC15过表达细胞中不增殖。本研究描述了从腹泻样本中鉴定HuNoV蛋白质附着因子的尝试。本研究结果将有助于理解HuNoV在人体内的感染、从各种环境中富集GII.4 HuNoV以及构建用于体外培养HuNoV的细胞系。
