title: Deletion of the non-essential UL0 gene of infectious laryngotracheitis (ILT) virus leads to attenuation in chickens, and UL0 mutants expressing influenza virus haemagglutinin (H7) protect against ILT and fowl plague.
abstract: Infectious laryngotracheitis virus (ILTV), a member of the Alphaherpesvirinae, possesses several unique genes. One of them, UL0, encodes an abundantly expressed protein that accumulates in the nuclei of ILTV-infected cells. This study demonstrates that this protein is dispensable for in vitro virus replication and that UL0 deletion mutants exhibit only minor growth defects in cultured cells. The UL0 gene locus of ILTV was also used for insertion of foreign DNA sequences encoding enhanced GFP or haemagglutinin (HA), subtype H7, of a highly pathogenic avian influenza virus under the control of the human cytomegalovirus immediate-early gene promoter. Expression of foreign proteins was shown by (immuno)fluorescence tests and Western blot analyses. After experimental infection of chickens, UL0 deletion mutants proved to be attenuated when compared to both parental wild-type ILTV and an UL0 rescue mutant. Nevertheless, all animals immunized with UL0-negative ILTV were protected from clinical disease after subsequent infection with virulent ILTV. Furthermore, all animals immunized with HA-expressing ILTV survived a lethal challenge with H7 subtype avian influenza virus with minimal clinical signs. Thus, an UL0-negative and HA-expressing ILTV recombinant may be used as a bivalent live virus vaccine against ILT and fowl plague. Unlike inactivated influenza virus vaccines, HA-expressing ILTV recombinants should be suitable for mass application and would also permit serological discrimination between vaccinated and virus-infected animals in the field.
authors.affiliation: Institutes of Molecular Biology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Jutta
authors.initials: J
authors.lastname: Veits
authors.affiliation: Institutes of Molecular Biology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Dörte
authors.initials: D
authors.lastname: Lüschow
authors.affiliation: Institutes of Molecular Biology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kindermann
authors.affiliation: Institutes of Diagnostic Virology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Ortrud
authors.initials: O
authors.lastname: Werner
authors.affiliation: Institutes of Infectology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Jens P
authors.initials: JP
authors.lastname: Teifke
authors.affiliation: Institutes of Molecular Biology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
authors.affiliation: Institutes of Molecular Biology, Friedrich-Loeffler Institutes, Federal Research Centre for Virus Diseases of Animals, D-17493 Greifswald-Insel Riems, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Fuchs
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002645:Chickens; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D006566:Herpesviridae Infections / Q000517:prevention & control / Q000662:veterinary*; D001352:Herpesvirus 1, Gallid / Q000235:genetics / Q000302:isolation & purification; D005585:Influenza in Birds / Q000517:prevention & control*; D007827:Laryngitis / Q000517:prevention & control / Q000662:veterinary*; D011200:Poultry; D011994:Recombinant Proteins / Q000096:biosynthesis; D014136:Tracheitis / Q000517:prevention & control / Q000662:veterinary*; D014611:Vaccination; D014613:Vaccines, Attenuated / Q000008:administration & dosage; D014614:Vaccines, Synthetic / Q000008:administration & dosage; D014765:Viral Vaccines / Q000008:administration & dosage*
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; C109963:hemagglutinin, avian influenza A virus