Rescue from cloned cDNAs and in vivo characterization of recombinant pathogenic Romero and live-attenuated Candid #1 strains of Junin virus, the causative agent of Argentine hemorrhagic fever disease.
The New World arenavirus Junin virus (JUNV) is the causative agent of Argentine hemorrhagic fever (AHF), which is associated with high morbidity and significant mortality. Several pathogenic strains of JUNV have been documented, and a highly attenuated vaccine strain (Candid #1) was generated and used to vaccinate the human population at risk. The identification and functional characterization of viral genetic determinants associated with AHF and Candid #1 attenuation would contribute to the elucidation of the mechanisms contributing to AHF and the development of better vaccines and therapeutics. To this end, we used reverse genetics to rescue the pathogenic Romero and the attenuated Candid #1 strains of JUNV from cloned cDNAs. Both recombinant Candid #1 (rCandid #1) and Romero (rRomero) had the same growth properties and phenotypic features in cultured cells and in vivo as their corresponding parental viruses. Infection with rRomero caused 100% lethality in guinea pigs, whereas rCandid #1 infection was asymptomatic and provided protection against a lethal challenge with Romero. Notably, Romero and Candid #1 trans-acting proteins, L and NP, required for virus RNA replication and gene expression were exchangeable in a minigenome rescue assay. These findings support the feasibility of studies aimed at determining the contribution of each viral gene to JUNV pathogenesis and attenuation. In addition, we rescued Candid #1 viruses with three segments that efficiently expressed foreign genes introduced into their genomes. This finding opens the way for the development of a safe multivalent arenavirus vaccine.
Department of Immunology and Microbial Science IMM-6, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Sebastien F
SF
Emonet
Alexey V
AV
Seregin
Nadezhda E
NE
Yun
Allison L
AL
Poussard
Aida G
AG
Walker
Juan C
JC
de la Torre
Slobodan
S
Paessler
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001117:Arenaviridae Infections / Q000276:immunology / Q000473:pathology / Q000517:prevention & control / Q000821:virology; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D005838:Genotype; D006168:Guinea Pigs; D006478:Hemorrhagic Fever, American / Q000276:immunology* / Q000473:pathology* / Q000517:prevention & control / Q000821:virology; D006801:Humans; D007114:Immunization; D018051:Junin virus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity* / Q000502:physiology; D008969:Molecular Sequence Data; D010641:Phenotype; D011995:Recombination, Genetic; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014779:Virus Replication
D000914:Antibodies, Viral; D018076:DNA, Complementary; D014613:Vaccines, Attenuated; D014765:Viral Vaccines