title: Permissible variation in the 3' non-coding region of the haemagglutinin genome segment of the H5N1 candidate influenza vaccine virus NIBRG-14 [corrected].
abstract: The candidate H5N1 vaccine virus NIBRG-14 was created in response to a call from the World Health Organisation in 2004 to prepare candidate vaccine viruses (CVVs) to combat the threat of an H5N1 pandemic. NIBRG-14 was created by reverse genetics and is composed of the neuraminidase (NA) and modified haemagglutinin (HA) genes from A/Vietnam/1194/2004 and the internal genes of PR8, a high growing laboratory adapted influenza A(H1N1) strain. Due to time constraints, the non-coding regions (NCRs) of A/Vietnam/1194/2004 HA were not determined prior to creating NIBRG-14. Consequently, the sequence of the primers used to clone the modified A/Vietnam/1194/2004 HA was based upon previous experience of cloning H5N1 viruses. We report here that the HA 3' NCR sequence of NIBRG-14 is different to that of the parental wild type virus A/Vietnam/1194/2004; however this does not appear to impact on its growth or antigen yield. We introduced additional small changes into the 3'NCR of NIBRG-14; these had only minor effects on viral growth and antigen content. These findings may serve to assure the influenza vaccine community that generation of CVVs using best-guess NCR sequences, based on sequence alignments, are likely to produce robust viruses.
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Health Protection Agency, Blanche Lane, Potters Bar, Hertfordshire, United Kingdom.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Johnson
authors.forename: Michelle
authors.initials: M
authors.lastname: Hamill
authors.forename: Ruth
authors.initials: R
authors.lastname: Harvey
authors.forename: Carolyn
authors.initials: C
authors.lastname: Nicolson
authors.forename: James S
authors.initials: JS
authors.lastname: Robertson
authors.forename: Othmar G
authors.initials: OG
authors.lastname: Engelhardt
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D004285:Dogs; D005818:Genetic Engineering; D014644:Genetic Variation; D016679:Genome, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity / Q000502:physiology; D007252:Influenza Vaccines / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D014776:Virus Cultivation; D014779:Virus Replication
chemical_list: D020413:3' Untranslated Regions; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D012367:RNA, Viral; D014614:Vaccines, Synthetic