title: Expression of a foreign epitope on infectious pancreatic necrosis virus VP2 capsid protein subviral particle (SVP) and immunogenicity in rainbow trout.
abstract: Infectious pancreatic necrosis virus (IPNV) is a major viral pathogen of salmonid fish and causes serious economic losses to salmonid aquaculture. Previously, we demonstrated that the IPNV capsid protein, VP2, expressed in yeast self-assembles into subviral particles (SVPs) and injection of these IPNV rVP2 SVPs into rainbow trout elicits an immune response. Immunized fish had reduced viral loads compared to unimmunized fish when challenged with IPNV. To evaluate the suitability of IPNV rVP2 SVPs for future development of multivalent vaccines, a linear epitope of a human oncogene, c-myc, was cloned into the IPNV rVP2 SVP backbone as a model epitope and expressed in yeast. Western blot analyses with anti-c-myc and anti-IPNV antibodies provided positive identification of both the c-myc and VP2 epitopes on the c-myc VP2 SVPs. Transmission electron microscopy of purified chimeric c-myc VP2 SVPs revealed the formation of approximately 20nm particles. Rainbow trout immunized with c-myc VP2 SVPs elicited both anti-c-myc and anti-IPNV immune responses. When immunized fish were challenged with IPNV, the viral load in the c-myc VP2 SVP immunized fish was significantly lower than the sham-vaccinated controls. The results indicate that IPNV rVP2 SVPs can tolerate the insertion of foreign epitopes without affecting either the antigenic potential of the epitopes of the backbone protein or the inserted foreign epitope. This opens the possibility of using the IPNV rVP2 SVP platform to express epitopes of other viruses, which could pave the way for development of multivalent subunit vaccines or novel marker vaccines.
authors.affiliation: Advanced BioNutrition Corp., 7155 Columbia Gateway Dr., Columbia, MD 21046, United States. adhar@abn-corp.com <adhar@abn-corp.com>
authors.forename: Arun K
authors.initials: AK
authors.lastname: Dhar
authors.forename: Robert M
authors.initials: RM
authors.lastname: Bowers
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Rowe
authors.forename: F C Thomas
authors.initials: FC
authors.lastname: Allnutt
mesh_terms: D000818:Animals; D018175:Birnaviridae Infections / Q000276:immunology; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005393:Fish Diseases / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D018061:Infectious pancreatic necrosis virus / Q000235:genetics*; D046529:Microscopy, Electron, Transmission; D017686:Oncorhynchus mykiss / Q000276:immunology*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology; D022721:Vaccines, Virosome / Q000008:administration & dosage / Q000276:immunology; D019562:Viral Load; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology*; D022701:Virosomes / Q000648:ultrastructure
chemical_list: D000939:Epitopes; D016271:Proto-Oncogene Proteins c-myc; C509831:VP2 protein, infectious pancreatic necrosis virus; D014614:Vaccines, Synthetic; D022721:Vaccines, Virosome; D015678:Viral Structural Proteins; D014765:Viral Vaccines; D022701:Virosomes