Banerjee Arinjay, Rapin Noreen, Miller Megan, Griebel Philip, Zhou Yan, Munster Vincent, Misra Vikram
Department of Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, MT, USA.
J Virol Methods. 2016 Nov;237:166-173. doi: 10.1016/j.jviromet.2016.09.008. Epub 2016 Sep 14.
It is speculated that bats are important reservoir hosts for numerous viruses, with 27 viral families reportedly detected in bats. Majority of these viruses have not been isolated and there is little information regarding their biology in bats. Establishing a well-characterized bat cell line supporting the replication of bat-borne viruses would facilitate the analysis of virus-host interactions in an in vitro model. Currently, few bat cell lines have been developed and only Tb1-Lu, derived from Tadarida brasiliensis is commercially available. Here we describe a method to establish and immortalize big brown bat (Eptesicus fuscus) kidney (Efk3) cells using the Myotis polyomavirus T-antigen. Subclones of this cell line expressed both epithelial and fibroblast markers to varying extents. Cell clones expressed interferon beta in response to poly(I:C) stimulation and supported the replication of four different viruses, namely, vesicular stomatitis virus (VSV), porcine epidemic diarrhea coronavirus (PED-CoV), Middle-East respiratory syndrome coronavirus (MERS-CoV) and herpes simplex virus (HSV). To our knowledge, this is the first bat cell line from a northern latitude insectivorous bat developed using a novel technology. The cell line has the potential to be used for isolation of bat viruses and for studying virus-bat interactions in culture.
据推测，蝙蝠是众多病毒的重要宿主，据报道在蝙蝠中检测到27个病毒科。这些病毒中的大多数尚未分离出来，关于它们在蝙蝠体内的生物学特性的信息也很少。建立一个能够支持蝙蝠携带病毒复制的特征明确的蝙蝠细胞系，将有助于在体外模型中分析病毒与宿主的相互作用。目前，已开发的蝙蝠细胞系很少，只有源自巴西无尾蝠的Tb1-Lu细胞系可商购。在此，我们描述了一种使用鼠耳蝠多瘤病毒T抗原建立并永生化大棕蝠（棕蝠）肾（Efk3）细胞的方法。该细胞系的亚克隆在不同程度上表达上皮和成纤维细胞标志物。细胞克隆在受到聚肌胞苷酸（poly(I:C)）刺激时表达β干扰素，并支持四种不同病毒的复制，即水泡性口炎病毒（VSV）、猪流行性腹泻冠状病毒（PED-CoV）、中东呼吸综合征冠状病毒（MERS-CoV）和单纯疱疹病毒（HSV）。据我们所知，这是首个利用新技术从北半球食虫蝙蝠建立的蝙蝠细胞系。该细胞系有潜力用于分离蝙蝠病毒以及研究培养中的病毒与蝙蝠的相互作用。
