Oelschlegel Anja M, Geissen Markus, Lenk Matthias, Riebe Roland, Angermann Marlies, Schatzl Herman, Groschup Martin H
Institute of Novel and Emerging Infectious Diseases at the Friedrich-Loeffler-Institut, Greifswald-Isle of Riems, Germany; Project Group Neuropharmacology, Leibniz Institute for Neurobiology, Magdeburg, Germany.
Institute of Novel and Emerging Infectious Diseases at the Friedrich-Loeffler-Institut, Greifswald-Isle of Riems, Germany; Department of Vascular Medicine, University Heart Centre Hamburg, UKE, Hamburg, Germany.
PLoS One. 2015 Jan 7;10(1):e0117154. doi: 10.1371/journal.pone.0117154. eCollection 2015.
Cell culture systems represent a crucial part in basic prion research; yet, cell lines that are susceptible to prions, especially to field isolated prions that were not adapted to rodents, are very rare. The purpose of this study was to identify and characterize a cell line that was susceptible to ruminant-derived prions and to establish a stable prion infection within it. Based on species and tissue of origin as well as PrP expression rate, we pre-selected a total of 33 cell lines that were then challenged with natural and with mouse propagated BSE or scrapie inocula. Here, we report the successful infection of a non-transgenic bovine cell line, a sub-line of the bovine kidney cell line MDBK, with natural sheep scrapie prions. This cell line retained the scrapie infection for more than 200 passages. Selective cloning resulted in cell populations with increased accumulation of PrPres, although this treatment was not mandatory for retaining the infection. The infection remained stable, even under suboptimal culture conditions. The resulting infectivity of the cells was confirmed by mouse bioassay (Tgbov mice, Tgshp mice). We believe that PES cells used together with other prion permissive cell lines will prove a valuable tool for ongoing efforts to understand and defeat prions and prion diseases.
细胞培养系统是朊病毒基础研究的关键部分;然而,对朊病毒敏感的细胞系,尤其是对未适应啮齿动物的野外分离朊病毒敏感的细胞系非常罕见。本研究的目的是鉴定和表征一种对反刍动物来源的朊病毒敏感的细胞系,并在其中建立稳定的朊病毒感染。基于物种、组织来源以及朊蛋白(PrP)表达率,我们预先选择了总共33种细胞系,然后用天然的以及小鼠传代的牛海绵状脑病(BSE)或羊瘙痒病接种物对其进行挑战。在此,我们报告了一种非转基因牛细胞系(牛肾细胞系MDBK的一个亚系)被天然羊瘙痒病朊病毒成功感染。该细胞系在200多次传代中保持了瘙痒病感染。选择性克隆导致PrPres积累增加的细胞群体,尽管这种处理对于维持感染并非必需。即使在次优培养条件下,感染仍保持稳定。通过小鼠生物测定(TgBov小鼠、TgShp小鼠)证实了细胞产生的感染性。我们相信,PES细胞与其他允许朊病毒感染的细胞系一起使用,将被证明是持续努力理解和战胜朊病毒及朊病毒疾病的有价值工具。