title: Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest.
abstract: Several arenaviruses cause hemorrhagic fever disease in humans for which no licensed vaccines are available and current therapeutic intervention is limited to the off-label use of the wide-spectrum antiviral ribavirin. However, the prototypic arenavirus lymphocytic choriomeningitis virus (LCMV) has proven to be a Rosetta stone for the investigation of virus-host interactions. Arenaviruses have a bisegmented negative-strand RNA genome. The S segment encodes for the virus nucleoprotein and glycoprotein, whereas the L segment encodes for the virus polymerase (L) and Z protein. The ability to generate recombinant LCMV (rLCMV) expressing additional foreign genes of interest would open novel avenues for the study of virus-host interactions and the development of novel vaccine strategies and high-throughput screens to identify antiarenaviral molecules. To this end, we have developed a trisegmented (1L + 2S) rLCMV-based approach (r3LCMV). Each of the two S segments in r3LCMV was altered to replace one of the viral genes by a gene of interest. All r3LCMVs examined expressing different reported genes were stable both genetically and phenotypically and exhibited wild-type growth properties in cultured cells. Reporter gene expression in r3LCMV-infected cells provided an accurate surrogate of levels of virus multiplication. Notably, some r3LCMVs displayed highly attenuated virulence in mice but induced protective immunity against a subsequent lethal challenge with wild-type LCMV, supporting the potential development of r3LCMV-based vaccines.
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Sebastien F
authors.initials: SF
authors.lastname: Emonet
authors.forename: Lucile
authors.initials: L
authors.lastname: Garidou
authors.forename: Dorian B
authors.initials: DB
authors.lastname: McGavern
authors.forename: Juan C
authors.initials: JC
authors.lastname: de la Torre
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004195:Disease Models, Animal; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D016679:Genome, Viral / Q000235:genetics*; D008216:Lymphocytic Choriomeningitis / Q000473:pathology / Q000821:virology; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D010641:Phenotype; D014771:Virion / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication