title: Attenuation of human respiratory syncytial virus by genome-scale codon-pair deoptimization.
abstract: Human respiratory syncytial virus (RSV) is the most important viral agent of serious pediatric respiratory-tract disease worldwide. A vaccine or generally effective antiviral drug is not yet available. We designed new live attenuated RSV vaccine candidates by codon-pair deoptimization (CPD). Specifically, viral ORFs were recoded by rearranging existing synonymous codons to increase the content of underrepresented codon pairs. Amino acid coding was completely unchanged. Four CPD RSV genomes were designed in which the indicated ORFs were recoded: Min A (NS1, NS2, N, P, M, and SH), Min B (G and F), Min L (L), and Min FLC (all ORFs except M2-1 and M2-2). Surprisingly, the recombinant CPD viruses were temperature-sensitive for replication in vitro (level of sensitivity: Min FLC > Min L > Min B > Min A). All of the CPD mutants grew less efficiently in vitro than recombinant wild-type (WT) RSV, even at the typically permissive temperature of 32 °C (growth efficiency: WT > Min L > Min A > Min FLC > Min B). CPD of the ORFs for the G and F surface glycoproteins provided the greatest restrictive effect. The CPD viruses exhibited a range of restriction in mice and African green monkeys comparable with that of two attenuated RSV strains presently in clinical trials. This study provided a new type of attenuated RSV and showed that CPD can rapidly generate vaccine candidates against nonsegmented negative-strand RNA viruses, a large and expanding group that includes numerous pathogens of humans and animals.
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and lenouenc@niaid.nih.gov Eckard.Wimmer@stonybrook.edu.
authors.forename: Cyril
authors.initials: C
authors.lastname: Le Nouën
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Linda G
authors.initials: LG
authors.lastname: Brock
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Cindy
authors.initials: C
authors.lastname: Luongo
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Thomas
authors.initials: T
authors.lastname: McCarty
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Yang
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Masfique
authors.initials: M
authors.lastname: Mehedi
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794 lenouenc@niaid.nih.gov Eckard.Wimmer@stonybrook.edu.
authors.forename: Eckard
authors.initials: E
authors.lastname: Wimmer
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794.
authors.forename: Steffen
authors.initials: S
authors.lastname: Mueller
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Peter L
authors.initials: PL
authors.lastname: Collins
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Ursula J
authors.initials: UJ
authors.lastname: Buchholz
authors.affiliation: RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
authors.forename: Joshua M
authors.initials: JM
authors.lastname: DiNapoli
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003062:Codon / Q000235:genetics*; D016679:Genome, Viral / Q000235:genetics*; D006801:Humans; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009305:Nasopharynx / Q000821:virology; D012367:RNA, Viral / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D013696:Temperature; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
chemical_list: D003062:Codon; D012367:RNA, Viral; D014764:Viral Proteins
keywords: live attenuated vaccine; negative strand RNA virus; pneumovirus