Institut für Virologie, Philipps-Universität Marburg, Marburg, Germany.
PLoS Negl Trop Dis. 2010 Aug 24;4(8):e802. doi: 10.1371/journal.pntd.0000802.
The fruit bat species Rousettus aegyptiacus was identified as a potential reservoir for the highly pathogenic filovirus Marburg virus. To establish a basis for a molecular understanding of the biology of filoviruses in the reservoir host, we have adapted a set of molecular tools for investigation of filovirus replication in a recently developed cell line, R06E, derived from the species Rousettus aegyptiacus.
METHODOLOGY/PRINCIPAL FINDINGS: Upon infection with Ebola or Marburg viruses, R06E cells produced viral titers comparable to VeroE6 cells, as shown by TCID(50) analysis. Electron microscopic analysis of infected cells revealed morphological signs of filovirus infection as described for human- and monkey-derived cell lines. Using R06E cells, we detected an unusually high amount of intracellular viral proteins, which correlated with the accumulation of high numbers of filoviral nucleocapsids in the cytoplasm. We established protocols to produce Marburg infectious virus-like particles from R06E cells, which were then used to infect naïve target cells to investigate primary transcription. This was not possible with other cell lines previously tested. Moreover, we established protocols to reliably rescue recombinant Marburg viruses from R06E cells.
CONCLUSION/SIGNIFICANCE: These data indicated that R06E cells are highly suitable to investigate the biology of filoviruses in cells derived from their presumed reservoir.
果蝠物种埃及果蝠被确定为高度致病性丝状病毒马尔堡病毒的潜在宿主。为了建立对储存宿主中丝状病毒生物学的分子理解基础，我们已经适应了一套分子工具，用于研究最近从物种埃及果蝠中开发的细胞系 R06E 中的丝状病毒复制。
方法/主要发现：用埃博拉或马尔堡病毒感染后，R06E 细胞产生的病毒滴度与 VeroE6 细胞相当，如 TCID（50）分析所示。感染细胞的电子显微镜分析显示出丝状病毒感染的形态学迹象，如人类和猴子来源的细胞系所描述的那样。使用 R06E 细胞，我们检测到异常高数量的细胞内病毒蛋白，这与细胞质中大量丝状核衣壳的积累相关。我们建立了从 R06E 细胞生产马尔堡传染性病毒样颗粒的方案，然后使用这些颗粒感染幼稚的靶细胞以研究初级转录。这在以前测试过的其他细胞系中是不可能的。此外，我们建立了从 R06E 细胞可靠地拯救重组马尔堡病毒的方案。
结论/意义：这些数据表明，R06E 细胞非常适合研究来自其假定宿主的细胞中丝状病毒的生物学。
