title: In vitro and in vivo immune responses induced by a DNA vaccine encoding the VP2 gene of the infectious pancreatic necrosis virus.
abstract: The work presented here describes the construction of a plasmid encoding the VP2 gene of the infectious pancreatic necrosis virus (IPNV), its expression in BF-2 cells and an evaluation of its activity in brown trout (Salmo trutta L) soon after injection. Preliminary experiments to evaluate the potential of the plasmid to induce neutralizing antibodies were also performed. We established a BF-2 cell line that expresses VP2 constitutively and we examined the infection of these VP2-transfected BF-2 cells with homologous and heterologous viruses. The expression kinetics of IFN, and of the IFN-induced genes Mx and ISG15, was also evaluated in brown trout over a 15 day interval, and quantified by real-time or semi-quantitative PCR. Type I IFN and Mx are markers of the non-specific innate immune response to viruses and they are involved in antiviral defences. Our results demonstrate that expression of the IPNV VP2 protein in BF-2 cells induces an antiviral state against IPNV and against the infectious haematopoietic necrosis virus (IHNV). In BF-2 infected cells, VP2 inhibited both the IPNV and IHNV-induced cytopathic effect to some extent, as well as the virus yield. In vivo, VP2 was expressed in haematopoietic tissues such as the head kidney of 7 month-old trout. In addition, it induced early immune responses and specific immunity 30 days after injection. IFN mRNA expression increased sharply on the 1st and 15th day post-injection and expression of other IFN-induced genes as Mx and ISG15 was also detected soon after vaccination of brown trout. Moreover, specific antibodies were detected 30 days after vaccination. These results suggest that the VP2 gene is a good candidate for the design of IPNV-DNA vaccines and to investigate the use of cytokines as co-stimulatory molecules.
authors.affiliation: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Departamento de Microbiología Molecular, C/Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Ana I
authors.initials: AI
authors.lastname: de Las Heras
authors.forename: Sara I
authors.initials: SI
authors.lastname: Pérez Prieto
authors.forename: Sylvia
authors.initials: S
authors.lastname: Rodríguez Saint-Jean
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018175:Birnaviridae Infections / Q000276:immunology* / Q000517:prevention & control; D002460:Cell Line; D005393:Fish Diseases / Q000276:immunology* / Q000517:prevention & control; D005786:Gene Expression Regulation / Q000276:immunology; D018061:Infectious pancreatic necrosis virus / Q000276:immunology*; D007668:Kidney / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014337:Trout / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology*; D015678:Viral Structural Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; C509831:VP2 protein, infectious pancreatic necrosis virus; D019444:Vaccines, DNA; D015678:Viral Structural Proteins; D014765:Viral Vaccines