title: Development of a subunit vaccine for infectious pancreatic necrosis virus using a baculovirus insect/larvae system.
abstract: Various attempts to develop a vaccine against infectious pancreatic necrosis virus (IPNV) have not yielded consistent results. Thus, at present, no commercial vaccine is available that can be used with confidence to immunize fry of salmon and trout. We generated a cDNA clone of the large genome segment A of an IPNV Sp strain and expressed all structural protein genes in insect cells and larvae using a baculovirus expression system. Green fluorescent protein was also coexpressed as a reporter molecule. High yields of IPNV proteins were obtained and the structural proteins self assembled to form virus-like particles (VLPs). We tested the immunogenicity of the putative VLP antigen in immersion vaccine experiments (two concentrations) in rainbow trout (Oncorhynchus mykiss) fry, and by intraperitoneal immunisation of Atlantic salmon (Salmo salar) pre-smolts using an oil adjuvant formulation. Rainbow trout were challenged by immersion using either the Sp or the VR-299 strain of IPNV two or three weeks post-vaccination, while Atlantic salmon were bath challenged with Sp strain after two months, after parr-smolt transformation. In the rainbow trout fry challenged two weeks post-immunization, cumulative mortality rates three weeks post challenge were 14 % in the fry that had received the highest dose versus 8 % in the control groups. No indication of protection was seen in repeated trials using a lower dose of antigen and challenge three weeks post-immunisation. The cumulative mortality rate of intraperitoneally immunised Atlantic salmon post-smolts four weeks post challenge was lower (56%) than in the control fish (77%), showing a dose-response pattern.
authors.affiliation: Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, MD 20742, USA.
authors.forename: R B
authors.initials: RB
authors.lastname: Shivappa
authors.forename: P E
authors.initials: PE
authors.lastname: McAllister
authors.forename: G H
authors.initials: GH
authors.lastname: Edwards
authors.forename: N
authors.initials: N
authors.lastname: Santi
authors.forename: O
authors.initials: O
authors.lastname: Evensen
authors.forename: V N
authors.initials: VN
authors.lastname: Vakharia
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D016367:Baculoviridae; D018175:Birnaviridae Infections / Q000517:prevention & control / Q000662:veterinary*; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000517:prevention & control* / Q000821:virology; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D018061:Infectious pancreatic necrosis virus / Q000235:genetics*; D012487:Salmonidae; D018411:Spodoptera; D014162:Transfection; D014611:Vaccination / Q000379:methods / Q000662:veterinary*; D022223:Vaccines, Subunit / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics*; D014771:Virion / Q000276:immunology / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D018076:DNA, Complementary; D022223:Vaccines, Subunit; D015678:Viral Structural Proteins; D014765:Viral Vaccines; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins