title: A T-cell epitope on NS3 non-structural protein enhances the B and T cell responses elicited by dendrimeric constructions against CSFV in domestic pigs.
abstract: It has been recently reported by our group that dendrimeric constructs combining B- and T-cell epitopes from classical swine fever virus (CSFV) provided partial protection against experimental infection. This research evaluated four newly designed constructions while taking into account our previous work, including the direct implication that a T-cell epitope from the NS3 protein contributes to the generation of the immune response against CSFV. To this end, the dendrimeric constructions, including either this NS3 T-cell epitope alone or two different B-cell epitopes without this T-cell epitope, were used to immunise pigs. Thus, construct 1, containing the NS3 T-cell epitope and four copies of a previously described B-cell epitope, significantly reduced the clinical scores and RNA viral loads after challenge relative to the control group. In three out of six animals in this group, vaccination achieved partial protection and was associated with IFN-gamma producing-cells and neutralising antibodies. In contrast, the pigs immunised with construct 2, again with four copies of the B epitope of construct 1 but lacking the T-cell motif, developed more severe clinical signs. Finally, the additional constructs 3 and 4 included four copies of a B epitope that was different from the epitope used in constructs 1 and 2 with or without the abovementioned NS3 T-cell epitope, respectively. Pigs immunised with these latter constructs developed low levels of peptide-specific antibodies that correlated with equally low levels of cellular responses, an absence of neutralising antibodies and a lack of protection. Even so, the clinical scores in the first week after the challenge were less severe for animals vaccinated with construct 3 than for those given construct 4. Our results confirm the relevant role of the B-cell epitope in residues 694-712 of the glycoprotein E2 (which is used in both constructs 1 and 2) for protection against CSFV, as well as the appropriateness of the newly used NS3 peptide as a specific T-cell epitope in domestic pigs.
authors.affiliation: Centre de Recerca en Sanitat Animal (CReSA), IRTA-UAB, Campus de la UAB, 08193 Bellaterra, Barcelona, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Tarradas
authors.forename: Marta
authors.initials: M
authors.lastname: Monsó
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Fraile
authors.forename: Beatriz G
authors.initials: BG
authors.lastname: de la Torre
authors.forename: Marta
authors.initials: M
authors.lastname: Muñoz
authors.forename: Rosa
authors.initials: R
authors.lastname: Rosell
authors.forename: Cristina
authors.initials: C
authors.lastname: Riquelme
authors.forename: Lester Josué
authors.initials: LJ
authors.lastname: Pérez
authors.forename: Miquel
authors.initials: M
authors.lastname: Nofrarías
authors.forename: Mariano
authors.initials: M
authors.lastname: Domingo
authors.forename: Francisco
authors.initials: F
authors.lastname: Sobrino
authors.forename: David
authors.initials: D
authors.lastname: Andreu
authors.forename: Llilianne
authors.initials: L
authors.lastname: Ganges
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D006691:Classical Swine Fever / Q000276:immunology* / Q000517:prevention & control; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000276:immunology*; D050091:Dendrimers / Q000138:chemical synthesis / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D018985:Epitopes, B-Lymphocyte / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D007114:Immunization / Q000662:veterinary; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000097:blood; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D018709:Statistics, Nonparametric; D013552:Swine; D013601:T-Lymphocytes / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000276:immunology*
chemical_list: D050091:Dendrimers; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D007074:Immunoglobulin G; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D007371:Interferon-gamma