title: A live RSV vaccine with engineered thermostability is immunogenic in cotton rats despite high attenuation.
abstract: Respiratory syncytial virus (RSV) is a leading cause of infant hospitalization and there remains no pediatric vaccine. RSV live-attenuated vaccines (LAVs) have a history of safe testing in infants; however, achieving an effective balance of attenuation and immunogenicity has proven challenging. Here we seek to engineer an RSV LAV with enhanced immunogenicity. Genetic mapping identifies strain line 19 fusion (F) protein residues that correlate with pre-fusion antigen maintenance by ELISA and thermal stability of infectivity in live RSV. We generate a LAV candidate named OE4 which expresses line 19F and is attenuated by codon-deoptimization of non-structural (NS1 and NS2) genes, deletion of the small hydrophobic (SH) gene, codon-deoptimization of the attachment (G) gene and ablation of the secreted form of G. OE4 (RSV-A2-dNS1-dNS2-ΔSH-dGm-Gsnull-line19F) exhibits elevated pre-fusion antigen levels, thermal stability, immunogenicity, and efficacy despite heavy attenuation in the upper and lower airways of cotton rats.
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Stobart
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Christina A
authors.initials: CA
authors.lastname: Rostad
authors.affiliation: School of Biology, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Zunlong
authors.initials: Z
authors.lastname: Ke
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Rebecca S
authors.initials: RS
authors.lastname: Dillard
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Cheri M
authors.initials: CM
authors.lastname: Hampton
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Strauss
authors.affiliation: Robert P Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, Georgia 30322, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Yi
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Anne L
authors.initials: AL
authors.lastname: Hotard
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Jia
authors.identifier: 0000-0002-1504-4335
authors.initials: J
authors.lastname: Meng
authors.affiliation: Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Raymond J
authors.initials: RJ
authors.lastname: Pickles
authors.affiliation: Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Kaori
authors.initials: K
authors.lastname: Sakamoto
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Sujin
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Currier
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20852, USA.
authors.forename: Syed M
authors.initials: SM
authors.lastname: Moin
authors.affiliation: Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20852, USA.
authors.forename: Barney S
authors.initials: BS
authors.lastname: Graham
authors.affiliation: Sigmovir Biosystems Inc., Rockville, Maryland 20850, USA.
authors.forename: Marina S
authors.initials: MS
authors.lastname: Boukhvalova
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Brian E
authors.initials: BE
authors.lastname: Gilbert
authors.affiliation: Sigmovir Biosystems Inc., Rockville, Maryland 20850, USA.
authors.forename: Jorge C G
authors.initials: JC
authors.lastname: Blanco
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas 77030, USA.|Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Pedro A
authors.initials: PA
authors.lastname: Piedra
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.|Robert P Apkarian Integrated Electron Microscopy Core, Emory University, Atlanta, Georgia 30322, USA.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Wright
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia 30322, USA.|Children's Healthcare of Atlanta, Atlanta, Georgia 30322, USA.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Moore
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D004355:Drug Stability; D006801:Humans; D008807:Mice, Inbred BALB C; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D018357:Respiratory Syncytial Virus Infections / Q000276:immunology* / Q000821:virology; D022261:Respiratory Syncytial Virus Vaccines / Q000235:genetics / Q000276:immunology*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D016205:Sigmodontinae; D013696:Temperature; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology*; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D022261:Respiratory Syncytial Virus Vaccines; D014613:Vaccines, Attenuated; D014764:Viral Proteins