title: Generation of biologically contained Ebola viruses.
abstract: Ebola virus (EBOV), a public health concern in Africa and a potential biological weapon, is classified as a biosafety level-4 agent because of its high mortality rate and the lack of approved vaccines and antivirals. Basic research into the mechanisms of EBOV pathogenicity and the development of effective countermeasures are restricted by the current biosafety classification of EBOVs. We therefore developed biologically contained EBOV that express a reporter gene instead of the VP30 gene, which encodes an essential transcription factor. A Vero cell line that stably expresses VP30 provides this essential protein in trans and biologically confines the virus to its complete replication cycle in this cell line. This complementation approach is highly efficient because biologically contained EBOVs lacking the VP30 gene grow to titers similar to those obtained with wild-type virus. Moreover, EBOVs lacking the VP30 gene are indistinguishable in their morphology from wild-type virus and are genetically stable, as determined by sequence analysis after seven serial passages in VP30-expressing Vero cells. We propose that this system provides a safe means to handle EBOV outside a biosafety level-4 facility and will stimulate critical studies on the EBOV life cycle as well as large-scale screening efforts for compounds with activity against this lethal virus.
authors.affiliation: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Halfmann
authors.forename: Jin Hyun
authors.initials: JH
authors.lastname: Kim
authors.forename: Hideki
authors.initials: H
authors.lastname: Ebihara
authors.forename: Takeshi
authors.initials: T
authors.lastname: Noda
authors.forename: Gabriele
authors.initials: G
authors.lastname: Neumann
authors.forename: Heinz
authors.initials: H
authors.lastname: Feldmann
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawaoka
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology; D002460:Cell Line; D002522:Chlorocebus aethiops; D004353:Drug Evaluation, Preclinical; D046129:Ebola Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D029043:Ebolavirus / Q000187:drug effects / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D006801:Humans; D010865:Pilot Projects; D012692:Serial Passage; D014157:Transcription Factors / Q000172:deficiency / Q000235:genetics; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics / Q000502:physiology*
chemical_list: D000998:Antiviral Agents; D046129:Ebola Vaccines; D014157:Transcription Factors; C479897:VP30 protein, ebola virus; D014764:Viral Proteins