Abstract
BackgroundThe 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 FindingsUpon infection with Ebola or Marburg viruses, R06E cells produced viral titers comparable to VeroE6 cells, as shown by TCID50 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/SignificanceThese data indicated that R06E cells are highly suitable to investigate the biology of filoviruses in cells derived from their presumed reservoir.
Highlights
Bats have been shown to be hosts for many pathogens, including those causing tropical diseases, such as leptospira, Hendra virus, Nipah virus and SARS-like coronavirus [1,2,3,4,5,6]
Marburg virus and several species of Ebola virus are endemic in central Africa and cause sporadic outbreaks in this region with mortality rates of up to 90%
As a first step towards understanding the biology of filoviruses in bats, we sought to establish a model system to investigate filovirus replication in cells derived from their natural reservoir
Summary
Bats have been shown to be hosts for many pathogens, including those causing tropical diseases, such as leptospira, Hendra virus, Nipah virus and SARS-like coronavirus [1,2,3,4,5,6]. Several species of fruit bats were identified as probable reservoirs for the filoviruses Marburg virus (MARV) and Ebola virus (EBOV) [7,8,9,10,11]. For more than 40 years the search for the natural reservoir of filoviruses was one of the most interesting endeavours in the field of highly pathogenic agents, and was fuelled by the dramatic outbreaks, cases of filovirus infected tourists and concerns that filoviruses might be abused as biological weapons. 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
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