title: Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine.
abstract: The poliovirus vaccine field is moving towards novel vaccination strategies. Withdrawal of the Oral Poliovirus Vaccine and implementation of the conventional Inactivated Poliovirus Vaccine (cIPV) is imminent. Moreover, replacement of the virulent poliovirus strains currently used for cIPV with attenuated strains is preferred. We generated Cold-Adapted Viral Attenuation (CAVA) poliovirus strains by serial passage at low temperature and subsequent genetic engineering, which contain the capsid sequences of cIPV strains combined with a set of mutations identified during cold-adaptation. These viruses displayed a highly temperature sensitive phenotype with no signs of productive infection at 37°C as visualized by electron microscopy. Furthermore, decreases in infectious titers, viral RNA, and protein levels were measured during infection at 37°C, suggesting a block in the viral replication cycle at RNA replication, protein translation, or earlier. However, at 30°C, they could be propagated to high titers (9.4-9.9 Log10TCID50/ml) on the PER.C6 cell culture platform. We identified 14 mutations in the IRES and non-structural regions, which in combination induced the temperature sensitive phenotype, also when transferred to the genomes of other wild-type and attenuated polioviruses. The temperature sensitivity translated to complete absence of neurovirulence in CD155 transgenic mice. Attenuation was also confirmed after extended in vitro passage at small scale using conditions (MOI, cell density, temperature) anticipated for vaccine production. The inability of CAVA strains to replicate at 37°C makes reversion to a neurovirulent phenotype in vivo highly unlikely, therefore, these strains can be considered safe for the manufacture of IPV. The CAVA strains were immunogenic in the Wistar rat potency model for cIPV, inducing high neutralizing antibody titers in a dose-dependent manner in response to D-antigen doses used for cIPV. In combination with the highly productive PER.C6 cell culture platform, the stably attenuated CAVA strains may serve as an attractive low-cost and (bio)safe option for the production of a novel next generation IPV.
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Barbara P
authors.initials: BP
authors.lastname: Sanders
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Isabel
authors.initials: I
authors.lastname: de Los Rios Oakes
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Vladimir
authors.initials: V
authors.lastname: van Hoek
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Viki
authors.initials: V
authors.lastname: Bockstal
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Tobias
authors.initials: T
authors.lastname: Kamphuis
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Taco G
authors.initials: TG
authors.lastname: Uil
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Yutong
authors.initials: Y
authors.lastname: Song
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Gillian
authors.initials: G
authors.lastname: Cooper
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Crawt
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Javier
authors.initials: J
authors.lastname: Martín
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Roland
authors.initials: R
authors.lastname: Zahn
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: John
authors.initials: J
authors.lastname: Lewis
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Eckard
authors.initials: E
authors.lastname: Wimmer
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Jerome H H V
authors.initials: JH
authors.lastname: Custers
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Hanneke
authors.initials: H
authors.lastname: Schuitemaker
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Jeronimo
authors.initials: J
authors.lastname: Cello
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, the Netherlands.
authors.forename: Diana
authors.initials: D
authors.lastname: Edo-Matas
mesh_terms: D000818:Animals; D003080:Cold Temperature; D006358:Hot Temperature; D008822:Mice, Transgenic; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D011051:Poliomyelitis / Q000276:immunology*; D017955:Poliovirus / Q000235:genetics / Q000276:immunology*; D011054:Poliovirus Vaccine, Inactivated / Q000276:immunology*; D011055:Poliovirus Vaccine, Oral / Q000276:immunology; D012367:RNA, Viral / Q000276:immunology; D051381:Rats; D014611:Vaccination / Q000379:methods
chemical_list: D011054:Poliovirus Vaccine, Inactivated; D011055:Poliovirus Vaccine, Oral; D012367:RNA, Viral