title: Recombinant infectious bursal disease virus carrying hepatitis C virus epitopes.
abstract: The delivery of foreign epitopes by a replicating nonpathogenic avian infectious bursal disease virus (IBDV) was explored. The aim of the study was to identify regions in the IBDV genome that are amenable to the introduction of a sequence encoding a foreign peptide. By using a cDNA-based reverse genetics system, insertions or substitutions of sequences encoding epitope tags (FLAG, c-Myc, or hepatitis C virus epitopes) were engineered in the open reading frames of a nonstructural protein (VP5) and the capsid protein (VP2). Attempts were also made to generate recombinant IBDV that displayed foreign epitopes in the exposed loops (P(BC) and P(HI)) of the VP2 trimer. We successfully recovered recombinant IBDVs expressing c-Myc and two different virus-neutralizing epitopes of human hepatitis C virus (HCV) envelope glycoprotein E in the VP5 region. Western blot analyses with anti-c-Myc and anti-HCV antibodies provided positive identification of both the c-Myc and HCV epitopes that were fused to the N terminus of VP5. Genetic analysis showed that the recombinants carrying the c-Myc/HCV epitopes maintained the foreign gene sequences and were stable after several passages in Vero and 293T cells. This is the first report describing efficient expression of foreign peptides from a replication-competent IBDV and demonstrates the potential of this virus as a vector.
authors.affiliation: University of Maryland Biotechnology Institute, Center of Marine Biotechnology, 701 East Pratt St., Baltimore, MD 21202-3101, USA.
authors.forename: Chitra
authors.initials: C
authors.lastname: Upadhyay
authors.forename: Arun
authors.initials: A
authors.lastname: Ammayappan
authors.forename: Deendayal
authors.initials: D
authors.lastname: Patel
authors.forename: Imre
authors.initials: I
authors.lastname: Kovesdi
authors.forename: Vikram N
authors.initials: VN
authors.lastname: Vakharia
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D004337:Drug Carriers; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007243:Infectious bursal disease virus / Q000235:genetics*; D007499:Iris Diseases; D016254:Mutagenesis, Insertional; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D017384:Sequence Deletion; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014761:Viral Hepatitis Vaccines / Q000235:genetics* / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D004337:Drug Carriers; D000939:Epitopes; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014761:Viral Hepatitis Vaccines