Arenavirus reverse genetics for vaccine development.
Arenaviruses are important human pathogens with no Food and Drug Administration (FDA)-licensed vaccines available and current antiviral therapy being limited to an off-label use of the nucleoside analogue ribavirin of limited prophylactic efficacy. The development of reverse genetics systems represented a major breakthrough in arenavirus research. However, rescue of recombinant arenaviruses using current reverse genetics systems has been restricted to rodent cells. In this study, we describe the rescue of recombinant arenaviruses from human 293T cells and Vero cells, an FDA-approved line for vaccine development. We also describe the generation of novel vectors that mediate synthesis of both negative-sense genome RNA and positive-sense mRNA species of lymphocytic choriomeningitis virus (LCMV) directed by the human RNA polymerases I and II, respectively, within the same plasmid. This approach reduces by half the number of vectors required for arenavirus rescue, which could facilitate virus rescue in cell lines approved for human vaccine production but that cannot be transfected at high efficiencies. We have shown the feasibility of this approach by rescuing both the Old World prototypic arenavirus LCMV and the live-attenuated vaccine Candid#1 strain of the New World arenavirus Junín. Moreover, we show the feasibility of using these novel strategies for efficient rescue of recombinant tri-segmented both LCMV and Candid#1.
Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Emilio
E
Ortiz-Riaño
Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Benson Yee Hin
BYH
Cheng
Department of Immunology and Microbial Science, Scripps Research Institute, La Jolla, CA 92037, USA.
Juan
J
Carlos de la Torre
Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Luis
L
Martínez-Sobrido
D000818:Animals; D001117:Arenaviridae Infections / Q000517:prevention & control / Q000821:virology*; D018050:Arenavirus / Q000235:genetics* / Q000276:immunology; D002522:Chlorocebus aethiops; D006224:Cricetinae; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D059386:Reverse Genetics / Q000379:methods*; D014709:Vero Cells; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology; D014776:Virus Cultivation
D014765:Viral Vaccines