title: In vitro and in vivo growth alter the population dynamic and properties of a Jeryl Lynn mumps vaccine.
abstract: Mumps vaccines are live attenuated viruses. They are known to vary in effectiveness, degree of attenuation and adverse event profile. However, the underlying reasons are poorly understood. We studied two closely related mumps vaccines which originate from the same attenuated Jeryl Lynn-5 strain but have different efficacies. Jeryl Lynn-Canine Kidney (JL-CK), produced on primary canine kidney cells, is less effective than RIT4385, which is produced on chicken embryo fibroblasts. JL-CK and RIT4385 could be distinguished by a number of in vitro and in vivo properties. JL-CK produced heterogeneous, generally smaller plaques than RIT4385, but gave 100-fold higher titres when grown in cells and showed a higher degree of hydrocephalus formation in neonatal rat brains. Sanger sequencing of JL-CK identified 14 regions of heterogeneity throughout the genome. Plaque purification of JL-CK demonstrated the presence of five different Jeryl Lynn-5 variants encompassing the 14 mutations. One JL-CK mutation was associated with a small plaque phenotype, the effects of the others in vitro or in vivo were less clear. Only 4% of the JL-CK population corresponded to the parental Jeryl Lynn-5 strain. Next generation sequencing of JL-CK and virus before and after growth in cell lines or neonatal rat brains showed that propagation in vitro or in vivo altered the population dramatically. Our findings indicate that growth of JL-CK in primary canine kidney cells resulted in the selection of a mixture of mumps virus variants that have different biological properties compared to the parent Jeryl Lynn-5 virus. We also report three previously unknown heterogenic regions within the N gene of the RIT4385 vaccine.
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom. Electronic address: Sarah.Gilliland@nibsc.org.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Connaughton
authors.affiliation: Laboratory for Molecular Structure, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom. Electronic address: Jun.Wheeler@nibsc.org.
authors.forename: Jun X
authors.initials: JX
authors.lastname: Wheeler
authors.affiliation: State Institute for Drug Control, Srobarova 48, 10041 Prague 10, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Vitková
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom. Electronic address: Philip.Minor@nibsc.org.
authors.forename: Philip
authors.initials: P
authors.lastname: Minor
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control (NIBSC), Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, United Kingdom. Electronic address: Silke.Schepelmann@nibsc.org.
authors.forename: Silke
authors.initials: S
authors.lastname: Schepelmann
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002642:Chick Embryo; D004195:Disease Models, Animal; D004847:Epithelial Cells; D006849:Hydrocephalus / Q000473:pathology / Q000821:virology; D009108:Mumps Vaccine / Q000008:administration & dosage / Q000276:immunology*; D009109:Mumps virus / Q000254:growth & development* / Q000276:immunology*; D011157:Population Dynamics; D051381:Rats; D013678:Technology, Pharmaceutical / Q000379:methods*; D019562:Viral Load; D010948:Viral Plaque Assay; D014774:Virulence; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D009108:Mumps Vaccine
keywords: Deep sequencing; Jeryl Lynn; Mumps virus; Neurovirulence; Vaccines