Zou Winnie Y, Blutt Sarah E, Crawford Sue E, Ettayebi Khalil, Zeng Xi-Lei, Saxena Kapil, Ramani Sasirekha, Karandikar Umesh C, Zachos Nicholas C, Estes Mary K
Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, MS-385, Houston, TX, 77030, USA.
Division of Gastroenterology and Hepatology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Methods Mol Biol. 2019;1576:229-247. doi: 10.1007/7651_2017_1.
Human rotavirus (HRV) and human norovirus (HuNoV) infections are recognized as the most common causes of epidemic and sporadic cases of gastroenteritis worldwide. The study of these two human gastrointestinal viruses is important for understanding basic virus-host interactions and mechanisms of pathogenesis and to establish models to evaluate vaccines and treatments. Despite the introduction of live-attenuated vaccines to prevent life-threatening HRV-induced disease, the burden of HRV illness remains significant in low-income and less-industrialized countries, and small animal models or ex vivo models to study HRV infections efficiently are lacking. Similarly, HuNoVs remained non-cultivatable until recently. With the advent of non-transformed human intestinal enteroid (HIE) cultures, we are now able to culture and study both clinically relevant HRV and HuNoV in a biologically relevant human system. Methods described here will allow investigators to use these new culture techniques to grow HRV and HuNoV and analyze new aspects of virus replication and pathogenesis.
人类轮状病毒(HRV)和人类诺如病毒(HuNoV)感染被认为是全球范围内流行性和散发性胃肠炎病例的最常见病因。对这两种人类胃肠道病毒的研究对于理解病毒与宿主的基本相互作用、发病机制以及建立评估疫苗和治疗方法的模型至关重要。尽管已引入减毒活疫苗来预防危及生命的HRV引起的疾病,但在低收入和工业化程度较低的国家,HRV疾病的负担仍然很重,并且缺乏能够有效研究HRV感染的小动物模型或体外模型。同样,直到最近HuNoV仍无法培养。随着未转化的人肠道类器官(HIE)培养技术的出现,我们现在能够在具有生物学相关性的人体系统中培养和研究具有临床相关性的HRV和HuNoV。本文所述的方法将使研究人员能够使用这些新的培养技术来培养HRV和HuNoV,并分析病毒复制和发病机制的新方面。