title: A DNA vaccine coding for the Brucella outer membrane protein 31 confers protection against B. melitensis and B. ovis infection by eliciting a specific cytotoxic response.
abstract: The development of an effective subunit vaccine against brucellosis is a research area of intense interest. The outer membrane proteins (Omps) of Brucella spp. have been extensively characterized as potential immunogenic and protective antigens. This study was conducted to evaluate the immunogenicity and protective efficacy of the B. melitensis Omp31 gene cloned in the pCI plasmid (pCIOmp31). Immunization of BALB/c mice with pCIOmp31 conferred protection against B. ovis and B. melitensis infection. Mice vaccinated with pCIOmp31 developed a very weak humoral response, and in vitro stimulation of their splenocytes with recombinant Omp31 did not induced the secretion of gamma interferon. Splenocytes from Omp31-vaccinated animals induced a specific cytotoxic-T-lymphocyte activity, which leads to the in vitro lysis of Brucella-infected macrophages. pCIOmp31 immunization elicited mainly CD8(+) T cells, which mediate cytotoxicity via perforins, but also CD4(+) T cells, which mediate lysis via the Fas-FasL pathway. In vivo depletion of T-cell subsets showed that the pCIOmp31-induced protection against Brucella infection is mediated predominantly by CD8(+) T cells, although CD4(+)T cells also contribute. Our results demonstrate that the Omp31 DNA vaccine induces cytotoxic responses that have the potential to contribute to protection against Brucella infection. The protective response could be related to the induction of CD8(+) T cells that eliminate Brucella-infected cells via the perforin pathway.
authors.affiliation: Laboratorio de Inmunogenética, Hospital de Clínicas José de San Martín, Facultad de Medicina, Universidad de Buenos Aires, Argentina. jucassat@ffyb.uba.ar
authors.forename: Juliana
authors.initials: J
authors.lastname: Cassataro
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Velikovsky
authors.forename: Silvia
authors.initials: S
authors.lastname: de la Barrera
authors.forename: Silvia M
authors.initials: SM
authors.lastname: Estein
authors.forename: Laura
authors.initials: L
authors.lastname: Bruno
authors.forename: Raúl
authors.initials: R
authors.lastname: Bowden
authors.forename: Karina A
authors.initials: KA
authors.lastname: Pasquevich
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Fossati
authors.forename: Guillermo H
authors.initials: GH
authors.lastname: Giambartolomei
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D002004:Brucella Vaccine / Q000235:genetics / Q000276:immunology; D017347:Brucella melitensis; D042061:Brucella ovis; D002006:Brucellosis / Q000276:immunology / Q000517:prevention & control*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D003001:Cloning, Molecular; D003602:Cytotoxicity, Immunologic; D053222:Fas Ligand Protein; D005260:Female; D005798:Genes, Bacterial; D007371:Interferon-gamma / Q000378:metabolism; D008264:Macrophages / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D002004:Brucella Vaccine; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; C101108:Omp31 protein, Brucella melitensis; D019444:Vaccines, DNA; D007371:Interferon-gamma