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Proc Natl Acad Sci U S A. 2014 Sep 9;111(36):13169-74. doi: 10.1073/pnas.1411290111. Epub 2014 Aug 25.

Attenuation of human respiratory syncytial virus by genome-scale codon-pair deoptimization.

Author information

1
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.
2
RNA Viruses Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; and.
3
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794 lenouenc@niaid.nih.gov Eckard.Wimmer@stonybrook.edu.
4
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794.

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.

KEYWORDS:

live attenuated vaccine; negative strand RNA virus; pneumovirus

PMID:
25157129
PMCID:
PMC4246931
DOI:
10.1073/pnas.1411290111
[Indexed for MEDLINE]
Free PMC Article

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