In vitro and in vivo characterization of recombinant Ebola viruses expressing enhanced green fluorescent protein.
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.
Ebihara
Hideki
H
Department of Special Pathogens, International Research Center for Infectious Diseases, University of Tokyo, Tokyo, Japan. Hideki_Ebihara@phac-aspc.gc.ca
Theriault
Steven
S
Neumann
Gabriele
G
Alimonti
Judie B
JB
Geisbert
Joan B
JB
Hensley
Lisa E
LE
Groseth
Allison
A
Jones
Steven M
SM
Geisbert
Thomas W
TW
Kawaoka
Yoshihiro
Y
Feldmann
Heinz
H
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
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins