Journal of Microbiology and Biotechnology
The Korean Society for Microbiology and Biotechnology publishes the Journal of Microbiology and Biotechnology.

2016 ; Vol.26-6: 1132~1139

AuthorYoung Bin Im, Woo Bin Park, Myunghwan Jung, Suk Kim, Han Sang Yoo
Place of dutyDepartment of Infectious Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea
TitleEvaluation of Th1/Th2-Related Immune Response against Recombinant Proteins of Brucella abortus Infection in Mice
PublicationInfo J. Microbiol. Biotechnol.2016 ; Vol.26-6
AbstractBrucellosis is a zoonotic disease caused by Brucella, a genus of gram-negative bacteria. Cytokines have key roles in the activation of innate and acquired immunities. Despite several research attempts to reveal the immune responses, the mechanism of Brucella infection remains unclear. Therefore, immune responses were analyzed in mice immunized with nine recombinant proteins. Cytokine production profiles were analyzed in the RAW 264.7 cells and naive splenocytes after stimulation with three recombinant proteins, metal-dependent hydrolase (r0628), bacterioferritin (rBfr), and thiamine transporter substrate-binding protein (rTbpA). Immune responses were analyzed by ELISA and ELISpot assay after immunization with proteins in mice. The production levels of NO, TNF-α, and IL-6 were time-dependently increased after having been stimulated with proteins in the RAW 264.7 cells. In naive splenocytes, the production of IFN-γ and IL-2 was increased after stimulation with the proteins. It was concluded that two recombinant proteins, r0628 and rTbpA, showed strong immunogenicity that was induced with Th1-related cytokines IFN-γ, IL-2, and TNF-α more than Th2-related cytokines IL-6, IL-4, and IL-5 in vitro. Conversely, a humoral immune response was activated by increasing the number of antigen-secreting cells specifically. Furthermore, these could be candidate diagnosis antigens for better understanding of brucellosis.
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Supplemental Data
Key_wordBrucella abortus, cytokine, immunogenicity, recombinant protein
References
  1. Al Dahouk S, Nockler K, Scholz HC, Tomaso H, Bogumil R, Neubauer H. 2006. Immunoproteomic characterization of Brucella abortus 1119-3 preparations used for the serodiagnosis of Brucella infection. J. Immunol. Methods 309: 34-47.
    Pubmed CrossRef
  2. Baldi PC, Giambartolomei GH, Goldbaum FA, Abdon LF, Velikovsky CA, Kittelberger R, Fossati CA. 1996. Humoral immune response against lipopolysaccharide and cytoplasmic proteins of Brucella abortus in cattle vaccinated with B. abortus S19 or experimentally infected with Yersinia enterocolitica serotype O:9. Clin. Diagn. Lab. Immunol. 3: 472-476.
    Pubmed Pubmed Central
  3. Berntsson RP, Smits SH, Schmitt L, Slotboom DJ, Poolman B. 2010. A structural classification of substrate-binding proteins. FEBS Lett. 584: 2606-2617.
    Pubmed CrossRef
  4. Cha SB, Rayamajhi N, Kang ML, Lee WJ, Shin MK, Yoo HS. 2010. Comparative study of gamma interferon production in mice immunized with outer membrane proteins and whole bacteria of Brucella abortus. Jpn. J. Infect. Dis. 63: 49-51.
    Pubmed
  5. Chaudhuri P, Prasad R, Kumar V, Gangaplara A. 2010. Recombinant OMP28 antigen-based indirect ELISA for serodiagnosis of bovine brucellosis. Mol. Cell. Probes 24: 142145.
    Pubmed CrossRef
  6. Cloeckaert A, Jacques I, Bosseray N, Limet JN, Bowden R, Dubray G, Plommet M. 1991. Protection conferred on mice by monoclonal antibodies directed against outer-membraneprotein antigens of Brucella. J. Med. Microbiol. 34: 175-180.
    Pubmed CrossRef
  7. Connolly JP, Comerci D, Alefantis TG, Walz A, Quan M, Chafin R, et al. 2006. Proteomic analysis of Brucella abortus cell envelope and identification of immunogenic candidate proteins for vaccine development. Proteomics 6: 3767-3780.
    Pubmed CrossRef
  8. Denoel PA, Crawford RM, Zygmunt MS, Tibor A, Weynants VE, Godfroid F, et al. 1997. Survival of a bacterioferritin deletion mutant of Brucella melitensis 16M in human monocytederived macrophages. Infect. Immun. 65: 4337-4340.
    Pubmed Pubmed Central
  9. Detilleux PG, Deyoe BL, Cheville NF. 1990. Entry and intracellular localization of Brucella spp. in Vero cells:fluorescence and electron microscopy. Vet. Pathol. 27: 317-328.
    Pubmed CrossRef
  10. Detilleux PG, Deyoe BL, Cheville NF. 1990. Penetration and intracellular growth of Brucella abortus in nonphagocytic cells in vitro. Infect. Immun. 58: 2320-2328.
    Pubmed Pubmed Central
  11. Fernández-Lago L, Monte M, Chordi A. 1996. Endogenous gamma interferon and interleukin-10 in Brucella abortus 2308 infection in mice. FEMS Immunol. Med. Microbiol. 15: 109-114
    Pubmed CrossRef
  12. Foster G, Osterman BS, Godfroid J, Jacques I, Cloeckaert A. 2007. Brucella ceti sp. nov. and Brucella pinnipedialis sp. nov. for Brucella strains with cetaceans and seals as their preferred host. Int. J. Syst. Evol. Microbiol. 57: 2688-2693.
    Pubmed CrossRef
  13. Franco MP, Mulder M, Gilman RH, Smith HL. 2007. Human brucellosis. Lancet Infect. Dis. 7: 775-786.
    CrossRef
  14. Ghasemi A, Jeddi-Tehrani M, Mautner J, Salari MH, Zarnani AH. 2014. Immunization of mice with a novel recombinant molecular chaperon confers protection against Brucella melitensis infection. Vaccine 32: 6659-6666.
    Pubmed CrossRef
  15. Ghasemi A, Zarnani AH, Ghoodjani A, Rezani S, Salari MH, Jeddi-Tehrani M. 2014. Identification of a new immunogenic candidate conferring protection against Brucella melitensis infection in mice. Mol. Immunol. 62: 142-149.
    Pubmed CrossRef
  16. Golding B, Scott DE, Scharf OL, Huang LY, Zaitseva M, Lapham C, et al. 2001. Immunity and protection against Brucella abortus. Microbes Infect. 3: 43-48.
    CrossRef
  17. González-Smith A, Vemulapalli R, Andrews E, Oñate A. 2006. Evaluation of Brucella abortus D NA vaccine b y expression of Cu-Zn superoxide dismutase antigen fused to IL-2. Immunobiology 211: 65-74.
    Pubmed CrossRef
  18. Guillermo HG, Zwerdling A, Cassataro J, Bruno L, Fossati CA, Philipp MT. 2004. Lipoprotein, not lipopolysaccharide, are the key mediators of proinflammatory response elicited by heat-killed Brucella abortus. J. Immunol. 173: 4635-4644.
    CrossRef
  19. Kindt TJ, Goldsby RA, Osborne BA. 2007. Kuby Immunology, pp. 306-307. 6th Ed. W.H. Freeman and Company, New York, NY.
  20. Kittelberger R, Bundesen PG, Cloeckaert A, Greiser-Wilke I, Letesson JJ. 1998. Serological cross-reactivity between Brucella abortus and Yersinia enterocolitica 0:9. Vet. Microbiol. 60: 45-51.
    CrossRef
  21. Ko J, Splitter GA. 2003. Molecular host-pathogen interaction in brucellosis: current understanding and future approaches to vaccine development for mice and humans. Clin. Microbiol. Rev. 16: 65-78.
    Pubmed CrossRef Pubmed Central
  22. Ko KY, Kim JW, Her M, Kang SI, Jung SC, Cho DH, Kim JY. 2012. Immunogenic proteins of Brucella abortus to minimize cross reactions in brucellosis diagnosis. Vet. Microbiol. 156:374-380.
    Pubmed CrossRef
  23. Lee JJ, Simborio HL, Reyes AW, Kim DG, Hop HT, Min W, et al. 2015. Immunoproteomic identification of immunodominant antigens independent of the time of infection in Brucella abortus 2308-challenged cattle. Vet. Res. 46: 17.
    Pubmed CrossRef Pubmed Central
  24. Lim JJ, Kim DH, Lee JJ, Kim DG, Min W, Lee HJ, et al. 2012. Evaluation of recombinant 28 kDa outer membrane protein of Brucella abortus for the clinical diagnosis of bovine brucellosis in Korea. J. Vet. Med. Sci. 74: 687-691.
    Pubmed CrossRef
  25. Nielsen K, Smith P, Yu WL, Halbert G. 2007. Salmonella enterica serotype Urbana interference with brucellosis serology. J. Immunoassay Immunochem. 28: 289-296.
    Pubmed CrossRef
  26. Okamura H, Kashiwamura S, Tsutsu H, Yoshimoto T, Nakanish K. 1998. Regulation of interferon-gamma production by IL-12 and IL-18. Curr. Opin. Immunol. 10: 259-264
    CrossRef
  27. Oñate AA, Vemulapalli R, Andrews E, Schurig GG, Boyle S, Folch H. 1999. Vaccination with live Escherichia coli expressing Brucella abortus Cu/Zn superoxide dismutase protects mice against virulent B. abortus. Infect. Immun. 67: 986-988.
    Pubmed Pubmed Central
  28. Ruiz-Mesa JD, Sanchez-Gonzalez J, Reguera JM, Maritin L, Lopez-Palmero S, Colmenero JD. 2005. Rose Bengal test:diagnostic yield and use for the rapid diagnosis of human brucellosis in emergency departments in endemic area. Clin. Microbiol. Infect. 11: 211-215.
    Pubmed CrossRef
  29. Scholz HC, Nöckler K, Göllner C, Bahn P, Vergnaud G, Tomaso H, et al. 2010. Brucella inopinata sp. nov., isolated from a breast implant infection. Int. J. Syst. Evol. Microbiol. 60: 801-808.
    Pubmed CrossRef
  30. Simborio HL, Lee JJ, Bernardo Reyes AW, Hop HT, Arayan LT, Min W, et al. 2015. Evaluation of the combined use of the recombinant Brucella abortus Omp10, Omp19 and Omp28 proteins for the clinical diagnosis of bovine brucellosis. Microb. Pathog. 83-84: 41-46.
    Pubmed CrossRef
  31. Sung KY, Jung M, Shin MK, Park HE, Lee JJ, Kim S, Yoo HS. 2014. Induction of immune responses by two recombinant proteins of Brucella abortus, outer membrane proteins 2b porin and Cu/Zn superoxide dismutase, in mouse model. J. Microbiol. Biotechnol. 24: 854-861.
    Pubmed CrossRef
  32. Wang M, Qureshi N, Soeurt N, Splitter G. 2001. High levels of nitric oxide production decrease early but increase rate survival of Brucella abortus in macrophage. Microb. Pathog. 31: 221-230.
    Pubmed CrossRef
  33. Whatmore AM, Davison N, Cloeckaert A, Al Dahouk S, Zygmunt MS, Brew SD, et al. 2014. Brucella papionis sp. nov., isolated from baboons (Papio spp.). Int. J. Syst. Evol. Microbiol. 64: 4120-4128.
    Pubmed CrossRef Pubmed Central
  34. Zhan Y, Kelso A, Cheers C. 1993. Cytokine production in the murine response to Brucella infection or immunization with antigenic extracts. Immunology 80: 458-464.
    Pubmed Pubmed Central



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