Ehlen Lukas, Tödtmann Jan, Specht Sabine, Kallies René, Papies Jan, Müller Marcel A, Junglen Sandra, Drosten Christian, Eckerle Isabella
Institute of Virology, University of Bonn Medical Centre, Sigmund-Freud-Strasse 25, 53127, Bonn, Germany.
Institute for Medical Microbiology, Immunology & Parasitology (IMMIP), University of Bonn Medical Centre, Sigmund-Freud-Strasse 25, 53127, Bonn, Germany.
Virol J. 2016 May 4;13:74. doi: 10.1186/s12985-016-0531-5.
Small mammals such as bats and rodents have been increasingly recognized as reservoirs of novel potentially zoonotic pathogens. However, few in vitro model systems to date allow assessment of zoonotic viruses in a relevant host context. The cotton rat (Sigmodon hispidus) is a New World rodent species that has a long-standing history as an experimental animal model due to its unique susceptibility to human viruses. Furthermore, wild cotton rats are associated with a large variety of known or potentially zoonotic pathogens.
A method for the isolation and culture of airway epithelial cell lines recently developed for bats was applied for the generation of rodent airway and renal epithelial cell lines from the cotton rat. Continuous cell lines were characterized for their epithelial properties as well as for their interferon competence. Susceptibility to members of zoonotic Bunya-, Rhabdo-, and Flaviviridae, in particular Rift Valley fever virus (RVFV), vesicular stomatitis virus (VSV), West Nile virus (WNV), and tick-borne encephalitis virus (TBEV) was tested. Furthermore, novel arthropod-derived viruses belonging to the families Bunya-, Rhabdo-, and Mesoniviridae were tested.
We successfully established airway and kidney epithelial cell lines from the cotton rat, and characterized their epithelial properties. Cells were shown to be interferon-competent. Viral infection assays showed high-titre viral replication of RVFV, VSV, WNV, and TBEV, as well as production of infectious virus particles. No viral replication was observed for novel arthropod-derived members of the Bunya-, Rhabdo-, and Mesoniviridae families in these cell lines.
In the current study, we showed that newly established cell lines from the cotton rat can serve as host-specific in vitro models for viral infection experiments. These cell lines may also serve as novel tools for virus isolation, as well as for the investigation of virus-host interactions in a relevant host species.
蝙蝠和啮齿动物等小型哺乳动物越来越被认为是新型潜在人畜共患病原体的宿主。然而,迄今为止,很少有体外模型系统能够在相关宿主环境中评估人畜共患病毒。棉鼠(Sigmodon hispidus)是一种新大陆啮齿动物,由于其对人类病毒具有独特的易感性,长期以来一直作为实验动物模型。此外,野生棉鼠与多种已知或潜在的人畜共患病原体有关。
将最近为蝙蝠开发的气道上皮细胞系分离和培养方法应用于从棉鼠中生成啮齿动物气道和肾上皮细胞系。对连续细胞系的上皮特性及其干扰素能力进行了表征。测试了其对人畜共患布尼亚病毒科、弹状病毒科和黄病毒科成员的易感性,特别是裂谷热病毒(RVFV)、水疱性口炎病毒(VSV)、西尼罗河病毒(WNV)和蜱传脑炎病毒(TBEV)。此外,还测试了属于布尼亚病毒科、弹状病毒科和中病毒科的新型节肢动物源性病毒。
我们成功地从棉鼠中建立了气道和肾上皮细胞系,并表征了它们的上皮特性。细胞显示具有干扰素能力。病毒感染试验显示RVFV、VSV、WNV和TBEV的高滴度病毒复制,以及感染性病毒颗粒的产生。在这些细胞系中未观察到布尼亚病毒科、弹状病毒科和中病毒科新型节肢动物源性成员的病毒复制。
在本研究中,我们表明新建立的棉鼠细胞系可作为病毒感染实验的宿主特异性体外模型。这些细胞系还可作为病毒分离的新工具,以及在相关宿主物种中研究病毒-宿主相互作用的工具。