Efficient strategy to generate a vectored duck enteritis virus delivering envelope of duck Tembusu virus

Viruses. 2014 Jun 20;6(6):2428-43. doi: 10.3390/v6062428.

Abstract

Duck Tembusu virus (DTMUV) is a recently emerging pathogenic flavivirus that has resulted in a huge economic loss in the duck industry. However, no vaccine is currently available to control this pathogen. Consequently, a practical strategy to construct a vaccine against this pathogen should be determined. In this study, duck enteritis virus (DEV) was examined as a candidate vaccine vector to deliver the envelope (E) of DTMUV. A modified mini-F vector was inserted into the SORF3 and US2 gene junctions of the attenuated DEV vaccine strain C-KCE genome to generate an infectious bacterial artificial chromosome (BAC) of C-KCE (vBAC-C-KCE). The envelope (E) gene of DTMUV was inserted into the C-KCE genome through the mating-assisted genetically integrated cloning (MAGIC) strategy, resulting in the recombinant vector, pBAC-C-KCE-E. A bivalent vaccine C-KCE-E was generated by eliminating the BAC backbone. Immunofluorescence and western blot analysis results indicated that the E proteins were vigorously expressed in C-KCE-E-infected chicken embryo fibroblasts (CEFs). Duck experiments demonstrated that the insertion of the E gene did not alter the protective efficacy of C-KCE. Moreover, C-KCE-E-immunized ducks induced neutralization antibodies against DTMUV. These results demonstrated, for the first time, that recombinant C-KCE-E can serve as a potential bivalent vaccine against DEV and DTMUV.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology
  • Antibodies, Viral / blood
  • Antibodies, Viral / immunology
  • Chick Embryo
  • Ducks
  • Flavivirus / genetics
  • Flavivirus / immunology*
  • Gene Expression
  • Gene Order
  • Genetic Vectors / genetics*
  • Genetic Vectors / immunology*
  • Immunization
  • Mardivirus / genetics*
  • Mardivirus / immunology*
  • Poultry Diseases / mortality
  • Poultry Diseases / prevention & control
  • Vaccine Potency
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / immunology*
  • Viral Plaque Assay
  • Viral Vaccines / genetics
  • Viral Vaccines / immunology

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Viral Envelope Proteins
  • Viral Vaccines