title: Newcastle disease virus expressing H5 hemagglutinin gene protects chickens against Newcastle disease and avian influenza.
abstract: Newcastle disease virus (NDV)-expressing avian influenza virus (AIV) hemagglutinin (HA) of subtype H5 was constructed by reverse genetics. A cloned full-length copy of the genome of the lentogenic NDV strain Clone 30 was used for insertion of the ORF encoding the HA of the highly pathogenic AIV isolate A/chicken/Italy/8/98 (H5N2) in the intergenic region between the NDV fusion and hemagglutinin-neuraminidase (HN) genes. Remarkably, two species of HA transcripts were detected in cells infected with the resultant NDVH5. In a second recombinant (NDVH5m), a NDV transcription termination signal-like sequence located within the HA ORF was eliminated by silent mutations. Consequently, NDVH5m produced 2.7-fold more full-length HA transcripts, expressed higher levels of HA, and also incorporated more HA protein into its envelope than NDVH5. NDVH5m stably expressed the modified HA gene for 10 egg passages and both recombinants were found innocuous after intracerebral inoculation of 1-day-old chickens. Immunization of chickens with NDVH5m induced NDV- and AIVH5-specific antibodies and protected chickens against clinical disease after challenge with a lethal dose of velogenic NDV or highly pathogenic AIV, respectively. Remarkably, shedding of influenza virus was not observed. Furthermore, immunization with NDVH5m permitted serological discrimination of vaccinated and AIV field virus-infected animals based on antibodies against the nucleoprotein of AIV. Therefore, recombinant NDVH5m is suitable as a bivalent vaccine against NDV and AIV and may be used as marker vaccine for the control of avian influenza.
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Boddenblick 5a, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Jutta
authors.initials: J
authors.lastname: Veits
authors.forename: Dorothee
authors.initials: D
authors.lastname: Wiesner
authors.forename: Walter
authors.initials: W
authors.lastname: Fuchs
authors.forename: Bernd
authors.initials: B
authors.lastname: Hoffmann
authors.forename: Harald
authors.initials: H
authors.lastname: Granzow
authors.forename: Elke
authors.initials: E
authors.lastname: Starick
authors.forename: Egbert
authors.initials: E
authors.lastname: Mundt
authors.forename: Horst
authors.initials: H
authors.lastname: Schirrmeier
authors.forename: Teshome
authors.initials: T
authors.lastname: Mebatsion
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
authors.forename: Angela
authors.initials: A
authors.lastname: Römer-Oberdörfer
mesh_terms: D000818:Animals; D002460:Cell Line; D002645:Chickens; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism*; D053125:Influenza A Virus, H5N2 Subtype; D005585:Influenza in Birds / Q000235:genetics / Q000821:virology*; D016253:Microscopy, Immunoelectron; D008957:Models, Genetic; D009521:Newcastle Disease / Q000235:genetics / Q000821:virology*; D009522:Newcastle disease virus / Q000378:metabolism*; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000737:chemistry; D014158:Transcription, Genetic; D014162:Transfection; D014612:Vaccines; D014765:Viral Vaccines / Q000737:chemistry
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; D014612:Vaccines; D014765:Viral Vaccines; C109963:hemagglutinin, avian influenza A virus