title: In vitro and in vivo characterization of recombinant Ebola viruses expressing enhanced green fluorescent protein.
abstract: To facilitate an understanding of the molecular aspects of the pathogenesis of Zaire ebolavirus (ZEBOV) infection, we generated 2 different recombinant viruses expressing enhanced green fluorescent protein (eGFP) from additional transcription units inserted at different positions in the virus genome. These viruses showed in vitro phenotypes similar to that of wild-type ZEBOV (wt-ZEBOV) and were stable over multiple passages. Infection with one of the viruses expressing eGFP produced only mild disease in rhesus macaques, demonstrating a marked attenuation in this animal model. However, in mice lacking signal transducer and activator of transcription 1, both viruses expressing eGFP caused lethal cases of disease that were moderately attenuated, compared with that caused by wt-ZEBOV. In mice, viral replication could be easily tracked by the detection of eGFP-positive cells in tissues, by use of flow cytometry. These findings demonstrate that the incorporation of a foreign gene will attenuate ZEBOV in vivo but that these viruses still have potential for in vitro and in vivo research applications.
authors.affiliation: Department of Special Pathogens, International Research Center for Infectious Diseases, University of Tokyo, Tokyo, Japan. Hideki_Ebihara@phac-aspc.gc.ca
authors.forename: Hideki
authors.initials: H
authors.lastname: Ebihara
authors.forename: Steven
authors.initials: S
authors.lastname: Theriault
authors.forename: Gabriele
authors.initials: G
authors.lastname: Neumann
authors.forename: Judie B
authors.initials: JB
authors.lastname: Alimonti
authors.forename: Joan B
authors.initials: JB
authors.lastname: Geisbert
authors.forename: Lisa E
authors.initials: LE
authors.lastname: Hensley
authors.forename: Allison
authors.initials: A
authors.lastname: Groseth
authors.forename: Steven M
authors.initials: SM
authors.lastname: Jones
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Geisbert
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawaoka
authors.forename: Heinz
authors.initials: H
authors.lastname: Feldmann
mesh_terms: D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015023:Democratic Republic of the Congo; D029043:Ebolavirus / Q000235:genetics* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics*; D000882:Haplorhini; D006801:Humans; D008264:Macrophages / Q000821:virology; D011994:Recombinant Proteins / Q000032:analysis*; D011995:Recombination, Genetic; D014162:Transfection; D020298:U937 Cells / Q000821:virology; D014709:Vero Cells / Q000821:virology
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins