title: Establishment of a bovine herpesvirus 4 based vector expressing a secreted form of the bovine viral diarrhoea virus structural glycoprotein E2 for immunization purposes.
abstract: BACKGROUND
  The biological characteristics of BoHV-4 make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity, unlikely oncogenicity, the capability to accommodate large amounts of foreign genetic material, the ability to infect several cell types from different animal species, and the ability to maintain transgene expression in both undifferentiated and differentiated cells.
  
  
  RESULTS
  A recombinant bovine herpesvirus 4 (BoHV-4CMV-IgKE2-14 Delta TK) expressing an enhanced secreted form of the bovine viral diarrhea virus (BVDV) structural glycoprotein E2 (gE2-14), obtained by the removal of the putative transmembrane domain and addition of a 14 amino acids peptide at its carboxyl terminal and an immunoglobulin K signal peptide to the amino terminal, was successfully constructed using a Recombineering (recombination -mediated genetic engineering) approach on BoHV-4 cloned as bacterial artificial chromosome. The galactokinase - based recombineering system was modified by the introduction of a kanamycin expression cassette and a kanamycin selection step that allowed a significant reduction of the untargeted background clones. BoHV-4CMV-IgKE2-14 Delta TK infected cell lines highly expressed gE2-14, which maintained native antigenic properties in a serum neutralization inhibition test. When rabbits and sheep were immunized with BoHV-4CMV-IgKE2-14 Delta TK, high levels of serum neutralized antibodies against BVDV were generated.
  
  
  CONCLUSIONS
  This work highlights the engineerization of BoHV-4 genome as a vector for vaccine purposes and may provide the basis for BVDV vaccination exploiting the BoHV-4- based vector that delivers an improved secreted version of the BVDV structural glycoprotein E2.
authors.affiliation: Università di Parma, Facoltà di Medicina Veterinaria, Dipartimento di Salute Animale, Sezione di Malattie Infettive degli Animali, via del Taglio 8, 43100 Parma, Italy. gaetano.donofrio@unipr.it
authors.forename: Gaetano
authors.initials: G
authors.lastname: Donofrio
authors.forename: Chiara
authors.initials: C
authors.lastname: Sartori
authors.forename: Lara
authors.initials: L
authors.lastname: Ravanetti
authors.forename: Sandro
authors.initials: S
authors.lastname: Cavirani
authors.forename: Laurent
authors.initials: L
authors.lastname: Gillet
authors.forename: Alain
authors.initials: A
authors.lastname: Vanderplasschen
authors.forename: Simone
authors.initials: S
authors.lastname: Taddei
authors.forename: Cesidio Filippo
authors.initials: CF
authors.lastname: Flammini
mesh_terms: D000818:Animals; D001908:Diarrhea Viruses, Bovine Viral / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics*; D026021:Herpesvirus 4, Bovine / Q000235:genetics*; D007114:Immunization / Q000379:methods*; D011817:Rabbits; D012756:Sheep; D014162:Transfection / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D014759:Viral Envelope Proteins