title: Characterization of culture supernatant proteins from Brucella abortus and its protection effects against murine brucellosis.
abstract: In this study, we characterized the secreted proteins of Brucella abortus into the enriched media under the bacterial laboratory growth condition and investigated the pathogenic importance of culture supernatant (CS) proteins to B. abortus infection. CS proteins from stationary phase were concentrated and analyzed using 2D electrophoresis. In MALDI TOF/TOF analysis, more than 27 proteins including CuZn SOD, Dps, Tat, OMPs, Adh, LivF, Tuf, SucC, GroEL and DnaK were identified. Cytotoxic effects of CS proteins were found to increase in a dose-dependent manner in RAW 264.7 cells. Upon B. abortus challenge into phagocytes, however, CS proteins pre-treated cells exhibited lower bacterial uptake and intracellular replication compared to untreated cells. Immunization with CS proteins induced a strong humoral and cell mediated immune responses and exhibited significant higher degree of protection against virulence of B. abortus infection compared to mice immunized with Brucella broth protein (BBP). Taken together, these results indicate that B. abortus secreted a number of soluble immunogenic proteins under laboratory culture condition, which can promote antibody production resulted in enhancing host defense against to subsequently bacterial infection. Moreover, further analysis of CS proteins may help to understand the pathogenic mechanism of B. abortus infection and host-pathogen interaction.
authors.affiliation: Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do 430-757, Republic of Korea; College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Jin Ju
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Jeong Ju
authors.initials: JJ
authors.lastname: Lim
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Dae Geun
authors.initials: DG
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Hannah Leah
authors.initials: HL
authors.lastname: Simborio
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Dong Hyeok
authors.initials: DH
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Alisha Wehdnesday Bernardo
authors.initials: AW
authors.lastname: Reyes
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: WonGi
authors.initials: W
authors.lastname: Min
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Hu Jang
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: School of Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Dong Hee
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Hong Hee
authors.initials: HH
authors.lastname: Chang
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea; Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea. Electronic address: kimsuk@gnu.ac.kr.
authors.forename: Suk
authors.initials: S
authors.lastname: Kim
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D002003:Brucella abortus / Q000276:immunology* / Q000378:metabolism*; D002006:Brucellosis / Q000517:prevention & control*; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D007114:Immunization; D051379:Mice; D020543:Proteome; D040901:Proteomics
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D020543:Proteome
keywords: Brucella abortus; Culture supernatant proteins; Protection; Virulence