title: Expression of MPB83 from Mycobacterium bovis in Brucella abortus S19 induces specific cellular immune response against the recombinant antigen in BALB/c mice.
abstract: Immunodominant MPB83 antigen from Mycobacterium bovis was expressed as a chimeric protein fused to either β-galactosidase, outer membrane lipoprotein OMP19 or periplasmic protein BP26 in gram-negative Brucella abortus S19, in all cases driven by each gene's own promoter. All fusion proteins were successfully expressed and localized in the expected subcellular fraction. Moreover, OMP19-MPB83 was processed as a lipoprotein when expressed in B. abortus. Splenocytes from BALB/c mice immunized with the recombinant S19 strains carrying the genes coding for the heterologous antigens in replicative plasmids, showed equally specific INF-γ production in response to MPB83 stimulation. Association to the lipid moiety of OMP19 presented no advantage in terms of immunogenicity for MPB83. In contrast, fusion to BP26, which was encoded by an integrative plasmid, resulted in a weaker immune response. None of the constructions affected the survival rate or the infection pattern of Brucella. We concluded that B. abortus S19 is an appropriate candidate for the expression of M. bovis antigens both associated to the membrane or cytosolic fraction and may provide the basis for a future combined vaccine for bovine brucellosis and tuberculosis.
authors.affiliation: Instituto de Biotecnología, CICVyA, INTA-Castelar, N. Repetto y Los Reseros s/n (1686) Hurlingham, Buenos Aires, Argentina.
authors.forename: Julia V
authors.initials: JV
authors.lastname: Sabio y García
authors.forename: Fabiana
authors.initials: F
authors.lastname: Bigi
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Rossetti
authors.forename: Eleonora
authors.initials: E
authors.lastname: Campos
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D002004:Brucella Vaccine / Q000235:genetics / Q000276:immunology; D002003:Brucella abortus / Q000235:genetics* / Q000276:immunology; D002417:Cattle; D005260:Female; D005822:Genetic Vectors; D007111:Immunity, Cellular; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008074:Lipoproteins / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000235:genetics / Q000276:immunology*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000276:immunology; D032581:Tuberculosis Vaccines / Q000235:genetics / Q000276:immunology; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D000942:Antigens, Bacterial; C106040:BP26 protein, Brucella; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002004:Brucella Vaccine; D008074:Lipoproteins; C094904:MPB83 protein, Mycobacterium; D008565:Membrane Proteins; C097319:OMP19 protein, Brucella abortus; D011993:Recombinant Fusion Proteins; D032581:Tuberculosis Vaccines; D007371:Interferon-gamma; D001616:beta-Galactosidase