title: Induction of immune responses by two recombinant proteins of brucella abortus, outer membrane proteins 2b porin and Cu/Zn superoxide dismutase, in mouse model.
abstract: The diagnosis of Brucella abortus is mainly based on serological methods using antibody against LPS, which has diagnostic problems. Therefore, to solve this problem, we evaluated two proteins of B. abortus, Cu/Zn superoxide dismutase (SodC) and outer membrane proteins 2b porin (Omp2b). The genes were cloned and expressed in a pMAL system, and the recombinant proteins, rOmp2b and rSodC, were purified as fusion forms with maltosebinding protein. The identity of the proteins was confirmed by SDS-PAGE and Western blot analysis with sera of mice infected with B. abortus. Production of cytokines and nitric oxide (NO) was investigated in RAW 264.7 cells and mouse splenocytes after stimulation with the proteins. Moreover, cellular and humoral immune responses were investigated in BALB/c mice after immunization with the proteins. TNF-α, IL-6, and NO were significantly inducible in RAW 264.7 cells. Splenocytes of naive mice produced IFN-γ and IL-4 significantly by stimulation. Moreover, number of IgG, IFN-γ, and IL-4 producing cells were increased in immunized mice with the two proteins. Production of IgG and IgM with rOmp2b was higher than those with rSodC in immunized mice. These results suggest that the two recombinant proteins of B. abortus may be potential LPS-free proteins for diagnosis.
authors.affiliation: Department of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Kyung Yong
authors.initials: KY
authors.lastname: Sung
authors.forename: Myunghwan
authors.initials: M
authors.lastname: Jung
authors.forename: Min-Kyoung
authors.initials: MK
authors.lastname: Shin
authors.forename: Hyun-Eui
authors.initials: HE
authors.lastname: Park
authors.forename: Jin Ju
authors.initials: JJ
authors.lastname: Lee
authors.forename: Suk
authors.initials: S
authors.lastname: Kim
authors.forename: Han Sang
authors.initials: HS
authors.lastname: Yoo
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002003:Brucella abortus / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D002006:Brucellosis / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D018272:Porins / Q000235:genetics / Q000276:immunology*; D013482:Superoxide Dismutase / Q000235:genetics / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D015850:Interleukin-6; C096736:Omp2b protein, Brucella; D018272:Porins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D013482:Superoxide Dismutase