title: Immunological properties of recombinant classical swine fever virus NS3 protein in vitro and in vivo.
abstract: Classical swine fever (CSF) is a highly contagious and often fatal disease of pigs characterised by fever, severe leukopenia and haemorrhages. With vaccines having an importance in disease control, studies are seeking improved protein-based subunit vaccine against the virus (CSFV). In this respect, recombinant viral NS3 protein was analysed for its immunopotentiating capacity, particularly in terms of cytotoxic immune responses. NS3 was effective at inducing in vitro responses, quantified by lymphoproliferation, IFN-gamma ELISPOT, flow cytometric detection of activated T cell subsets, and cytotoxic T cell assays. Peripheral blood mononuclear cells from CSFV-immune pigs could be stimulated, but not cells from naïve animals. In addition to the IFN-gamma responses, induction of both CD4+ T helper cell and CD8+ cytotoxic T cells (CTL) were discernible--activation of the latter was confirmed in a virus-specific cytolytic assay. Attempts were made to translate this to the in vivo situation, by vaccinating pigs with an E2/NS3-based vaccine compared with an E2 subunit vaccine. Both vaccines were similar in their abilities to stimulate specific immune responses and protect pigs against lethal CSFV infection. Although the E2/NS3 vaccine appeared to have an advantage in terms of antibody induction, this was not statistically significant when group studies were performed. It was also difficult to visualise the NS3 capacity to promote T-cell responses in vivo. These results show that NS3 has potential for promoting cytotoxic defences, but the formulation of the vaccine requires optimisation for ensuring that NS3 is correctly delivered to antigen presenting cells for efficient activation of CTL.
authors.affiliation: Institute of Virology and Immunoprophylaxis, 3147 Mittelhäusern, Switzerland.
authors.forename: Henriette
authors.initials: H
authors.lastname: Rau
authors.forename: Hilde
authors.initials: H
authors.lastname: Revets
authors.forename: Carole
authors.initials: C
authors.lastname: Balmelli
authors.forename: Kenneth C
authors.initials: KC
authors.lastname: McCullough
authors.forename: Artur
authors.initials: A
authors.lastname: Summerfield
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000097:blood; D006691:Classical Swine Fever / Q000276:immunology / Q000517:prevention & control*; D006692:Classical Swine Fever Virus / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry / Q000662:veterinary; D007111:Immunity, Cellular / Q000276:immunology; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D009500:Neutralization Tests / Q000662:veterinary; D020365:RNA Helicases / Q000276:immunology; D011897:Random Allocation; D012697:Serine Endopeptidases / Q000276:immunology; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014611:Vaccination / Q000662:veterinary; D022223:Vaccines, Subunit / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000914:Antibodies, Viral; D007371:Interferon-gamma; D020365:RNA Helicases; D012697:Serine Endopeptidases; D022223:Vaccines, Subunit; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases