title: DNA vaccination with plasmids containing various fragments of large segment genome of infectious bursal disease virus.
abstract: The present study was undertaken to determine the effectiveness in including VP2 gene of the large segment genome of infectious bursal disease virus (IBDV) in the DNA vaccine for protection of chickens against infectious bursal disease (IBD). Different fragments of the large segment gene of IBDV standard challenge (STC) strain were successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR) followed by cloning into a eukaryotic expression plasmid vector (pCR3.1) as DNA vaccines. Transient expression of the encoded genes from various constructed plasmids was characterized in COS-7 cells by positive immunofluorescent staining with polyclonal or monoclonal antibody to IBDV. Chickens (1-day old) were intramuscularly injected with the individual plasmid three times at weekly intervals, challenged with IBDV strain STC at 21-day old, and observed for 10 days. Chickens receiving the plasmids containing VP2 genes, including VP243, VP24, or VP2 fragment, did not have clinical signs, mortality, and bursal atrophy and were effectively protected against IBDV infection. On the contrary, chickens receiving plasmids without containing VP2 genes, including VP4, VP3, or VP43 fragment, had marked bursal atrophy and were not protected against IBD. Antigen detection was correlated with protection; chickens protected from IBDV infection had undetectable IBDV antigen in bursae. Enzyme-linked immunosorbent assay (ELISA) antibody titer to IBDV was low or undetectable prior to or after challenge with IBDV in protected chickens receiving the plasmids containing the VP2 gene. The results indicate that inclusion of VP2 gene in the plasmid DNA is essential in achieving effective protection mediated by DNA vaccination against IBDV infection in chickens.
authors.affiliation: Department of Veterinary Pathobiology, 1175 ADDL, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Hua-Chen
authors.initials: HC
authors.lastname: Chang
authors.forename: Tsang-Long
authors.initials: TL
authors.lastname: Lin
authors.forename: Ching-Ching
authors.initials: CC
authors.lastname: Wu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D002060:Bursa of Fabricius / Q000276:immunology; D019556:COS Cells; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression / Q000235:genetics; D016678:Genome; D007243:Infectious bursal disease virus / Q000235:genetics / Q000276:immunology*; D010957:Plasmids / Q000276:immunology*; D014611:Vaccination; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D019444:Vaccines, DNA