title: A Reverse Genetics Approach for the Design of Methyltransferase-Defective Live Attenuated Avian Metapneumovirus Vaccines.
abstract: Avian metapneumovirus (aMPV), also known as avian pneumovirus or turkey rhinotracheitis virus, is the causative agent of turkey rhinotracheitis and is associated with swollen head syndrome in chickens. aMPV belongs to the family Paramyxoviridae which includes many important human pathogens such as human respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and human parainfluenza virus type 3 (PIV3). The family also includes highly lethal emerging pathogens such as Nipah virus and Hendra virus, as well as agriculturally important viruses such as Newcastle disease virus (NDV). For many of these viruses, there is no effective vaccine. Here, we describe a reverse genetics approach to develop live attenuated aMPV vaccines by inhibiting the viral mRNA cap methyltransferase. The viral mRNA cap methyltransferase is an excellent target for the attenuation of paramyxoviruses because it plays essential roles in mRNA stability, efficient viral protein translation and innate immunity. We have described in detail the materials and methods used to generate recombinant aMPVs that lack viral mRNA cap methyltransferase activity. We have also provided methods to evaluate the genetic stability, pathogenesis, and immunogenicity of live aMPV vaccine candidates in turkeys.
authors.affiliation: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 311 Goss Laboratory, 1925 Coffey Road, Columbus, OH, 43210, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 311 Goss Laboratory, 1925 Coffey Road, Columbus, OH, 43210, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 311 Goss Laboratory, 1925 Coffey Road, Columbus, OH, 43210, USA.
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 311 Goss Laboratory, 1925 Coffey Road, Columbus, OH, 43210, USA. li.926@osu.edu.
authors.forename: Jianrong
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D002645:Chickens / Q000821:virology; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D006801:Humans; D007700:Kinetics; D029121:Metapneumovirus / Q000201:enzymology / Q000235:genetics* / Q000276:immunology* / Q000502:physiology; D008745:Methylation; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009928:Organ Specificity; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D059386:Reverse Genetics / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014422:Turkeys / Q000821:virology; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics; D014765:Viral Vaccines / Q000235:genetics* / Q000276:immunology*; D014779:Virus Replication
chemical_list: D018076:DNA, Complementary; D014613:Vaccines, Attenuated; D014764:Viral Proteins; D014765:Viral Vaccines
keywords: Avian metapneumovirus; Methyltransferase; Reverse genetics; Vaccine