Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine
| Autor: | Lewis John Alfred, Javier Martin, Isabel de los Rios Oakes, Vladimir van Hoek, Viki Bockstal, Laura Crawt, Taco G. Uil, Gillian Cooper, Yutong Song, Eckard Wimmer, Diana Edo-Matas, Roland Zahn, Barbara Petronella Sanders, Hanneke Schuitemaker, Jerome Custers, Jeronimo Cello, Tobias Kamphuis |
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| Rok vydání: | 2016 |
| Předmět: |
RNA viruses
0301 basic medicine Hot Temperature Physiology Pathology and Laboratory Medicine medicine.disease_cause Biochemistry Enteroviruses Serial passage Immune Physiology Medicine and Health Sciences Public and Occupational Health CD155 Biology (General) Neutralizing antibody Internal Ribosome Entry Site Vaccines Immune System Proteins Attenuated vaccine Viral Vaccine Poliovirus Vaccination Microbial Mutation Vaccination and Immunization Cold Temperature Phenotype Medical Microbiology Viral Pathogens Viruses Inactivated Poliovirus Vaccine RNA Viral Pathogens Research Article Attenuated Vaccines QH301-705.5 Immunology Mice Transgenic Biology Microbiology 03 medical and health sciences Virology Genetics medicine Animals Antigens Microbial Pathogens Molecular Biology Organisms Biology and Life Sciences Proteins RC581-607 Viral Replication Rats Poliovirus Vaccine Inactivated 030104 developmental biology Viral replication Poliovirus Vaccine Oral Mutation biology.protein Parasitology Preventive Medicine Immunologic diseases. Allergy Poliomyelitis |
| Zdroj: | PLoS Pathogens, Vol 12, Iss 3, p e1005483 (2016) PLoS Pathogens |
| ISSN: | 1553-7374 |
| DOI: | 10.1371/journal.ppat.1005483 |
| Popis: | 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. Author Summary The vaccines that are used to protect against poliovirus infection have been available since the 1950s and have brought the eradication of poliomyelitis to our doorstep. For the post-eradication era, an Inactivated Poliovirus Vaccine (IPV) based on attenuated Sabin strains is recommended, as these strains are currently the only option to move to safer manufacturing of IPV. Here we describe three novel poliovirus strains that cannot replicate at 37°C. Their lack of pathogenicity was confirmed by intracerebral inoculation of susceptible transgenic mice that subsequently did not develop any symptoms of poliomyelitis. The inability to replicate at 37°C is caused by multiple mutations which do not revert to virulence after passage in cells. Furthermore, when used as vaccines, these viruses were capable of inducing a potent immune response in rats. At low temperature (30°C) these viruses showed high productivity on the PER.C6 cell line, which has the potential to significantly reduce costs of goods, as previously shown for conventional poliovirus strains. Taken together, these new strains could contribute to a safe, genetically stable, efficacious and affordable IPV. |
| Databáze: | OpenAIRE |
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