Related articles in JMB
Molecular and Cellular Microbiology (MCM) | Clinical Microbiology and Biomedical Sciences
Substantial Protective Immunity Conferred by a Combination of Brucella abortus Recombinant Proteins against Brucella abortus 544 Infection in BALB/c Mice
Gyeongsang National University, Republic of KoreaCorrespondence to:
Received: November 2, 2018; Accepted: December 20, 2018
J. Microbiol. Biotechnol. 2019; 29(2): 330-338
Published February 28, 2019
Copyright © The Korean Society for Microbiology and Biotechnology.
Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and DH5α, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with 5 × 104 CFU of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-γ is significantly elevated while IL-10, which is considered beneficial to the pathogen’s survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.
B. abortus, recombinant proteins, vaccination, cytokines, protection
- Hop HT, Simborio HL, Reyes AW, Arayan LT, Min W, Lee HJ. 2015. Immunogenicity and protective effect of recombinant Brucella abortus Ndk (rNdk) against a virulent strain Brucella abortus 544 infection in BALB/c mice. FEMS. Microbiol. Lett. 362: 1-6.
- Trujillo IZ, Zavala AN, Caceres JG, Miranda CQ. Brucellosis. 1994. Infect. Dis. Clin. North. Am. 8: 225-241.
- Franco MP, Mulder M, Gilman RH, Smits HL. Human brucellosis. 2007. Lance. Infect. Dis. 7: 775-786.
- 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.
- Corbel MJ. 1997. Brucellosis: an overview. Emerg. Infect. Dis. 3: 213-221.
- Sangari FJ, Aguero J. 1996. Molecular basis of Brucella pathogenicity: an update. Microbiologia 12: 207-218.
- Baldwin CL, Winter AJ. 1994. Macrophages and Brucella. Immunol. Ser. 60: 363-380.
- Ebel ED, Williams MS, Tomlinson SM. 2008. Estimating herd prevalence of bovine brucellosis in 46 USA states using slaughter surveillance. Prev. Vet. Med. 85: 295-316.
- Treanor JJ, Johnson JS, Wallen RL, Cilles S, Crowley PH, Cox JJ, et al. 2010. Vaccination strategies for managing brucellosis in Yellowstone bison. Vaccine 28: F64-F72.
- Galindo RC, de Miguel MJ, Labairu J, Marin CM, Revilla M, Blasco JM, et al. 2010. Gene expression changes in spleens of the wildlife reservoir species, Eurasian wild boar (Sus scrofa), naturally infected Brucella suis biovar 2. J. Genet. Genomics 37: 725-736.
- Olsen SC, Hennayer SG. 2010. Immune responses and protection against experimental Brucella suis biovar 1 challenge in non-vaccinated cattle. Clin. Vaccine Immunol. 17:1891-1895.
- Perrie Y, Mohammed AR, Kirby DJ, McNeil SE, Bramwell VW. 2008. Vaccine adjuvant systems: enhancing the efficacy of sub-unit protein antigens. Int. J. Pharm. 364: 272-280.
- Arena-Gamboa AM, Ficht TA, Kahl-McDonagh MM, RiceFicht AC. 2008. Immunization with a single dose of a microencapsulated Brucella melitensis m u atnt e nhances protection against wild-type challenge. Immun. Infect. 76:2448-255.
- Rafiei A, Ardestani SK, Kariminia A, Keyhani A, Mohraz M, Armithani A. 2006. Dominant Th1 cytokine production in early onset of human brucellosis followed by switching towards Th2 along prolongation of disease. J. Infect. 53: 315324.
- Reyes AWB, Simborio HLT, Huyhn TH, Arayan LT, Kim S. 2016. Molecular cloning, purification and immunogenicity of recombinant Brucella abortus 544 malate dehydrogenase protein. J. Vet. Sci. 17: 119-122.
- Velikovsky CA, Goldbaum FA, Cassataro J, Estein S, Bowden RA, Bruno L, et al. 2003. Brucella lumazine synthase elicits a mixed Th1-Th2 immune response and reduces infection in mice challenged with Brucella abortus 544 independently of the adjuvant formulation used. Infect. Immun. 71: 5750-5755.
- Stevens MG, Pugh G Jr, Tabatabai LB. 1992. Effects of gamma interferon and indomethacin in preventing Brucella abortus infections in mice. Infect. Immun. 60: 4407-4409.
- Oliveira SC, Splitter GA. 1996. Immunization of mice with recombinant L7/L12 ribosomal protein confers protection against Brucella abortus infection. Vaccine 14: 959-962.
- Dubray G, Bezard G. 1980. Isolation of three Brucella abortus cell wall antigens protective in murine experimental brucellosis. Ann. Rech. Vet. 11: 367-373.
- Lim JJ, Kim DH, Kim DG, Kim DG, Min W, Lee HJ, et al. 2012. Protective effects of recombinant Brucella abortus Omp28 against infection with a virulent strain of Brucella abortus 544 in mice. Vet. Sci. 13: 287-292.
- Miranda KL, Dorneles EMS, Pauletti RB, Poester FP, Lage AP. 2015. Brucella abortus S19 and RB51 vaccine immunogenicity test: evaluation of three mice (BALB/c, Swiss and CD-1) and two challenge strains (544 and 2308). Vaccine. 33: 507-511.
- Schurig GG, Sriranganathan N, Corbel MJ. 2002. Brucellosis vaccines: past, present and future. Vet. Microbiol. 90: 479496.
- Araya LN, Elzer PH, Rowe GE, Enright FM, Winter AJ. 1989. Temporal development of protective cell-mediated and humoral immunity in BALB/c mice infected with Brucella abortus. J. Immunol. 143: 3330-3337.
- Pavlov HM, Hogarth IF, McKenzie F, Cheers C. 1982. In vivo and in vitro effects of monoclonal antibody to Ly antigens on immunity to infection. Cell. Immunol. 71: 127-138.
- Truong QL, Cho Y, Kim K, Park BK, Hahn TW. 2015. Booster vaccination with safe, modified, live-attenuated mutants of Brucella abortus strain RB51 vaccine confers protective immunity against virulent strains of B. abortus and Brucella canis in BALB/c mice. Microbiology 161: 2137-2148.
- Bachrach G, Banai M, Bardenstein S, Hoida G, Genizi A, Bercovier H. 1994. Brucella ribosomal protein L7/L12 is a major component in the antigenicity of Brucella in INRA for delayed-type hypersensitivity in Brucella-sensitized guinea pigs. Infect. Immun. 62: 5361-5366.
- Gee J, Valderas MW, Kovach M, Grippe VK, Robertson GT, Ng WL, et al. 2005. The Brucella abortus Cu, Zn superoxide dismutase is required for optimal resistance to oxidative killing by murine macrophages and wild-type virulence in experimentally infected mice. Infect. Immun. 73: 2873-2880.
- Tabatai L B, P u gh G W. 1 994. M odu l ation o f immu ne responses in BALB/c mice vaccinated with Brucella abortus Cu-Zn superoxide dismutase synthetic peptide vaccine. Vaccine 12: 919-924.
- Piddington DL, Fang FC, Laessig T, Cooper AM, Orme IM, Buchmeier NA. 2001. Cu, Zn superoxide dismutase of Mycobacterium tuberculosis contributes to survival in activated macrophages that are generating an oxidative burst. Infect. Immun. 69: 4980-4987.
- De Groote MA, Ochner UA, Shiloh MU Nathan C, McCord JM, Dinauer MC, Libby SJ, et al. 1997. Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH oxidase and nitric oxide synthase. Proc. Natl. Acad. Sci. 94: 13997-14001.
- Velikovsky CA, Cassataro J, Giambartolomei GH, Goldbaum FA, Estein S, Bowden RA, et al. 2002. A DNA vaccine encoding lumazine synthase from Brucella abortus induces protective immunity in BALB/c mice. Infect. Immun. 70: 2507-2511.
- Oliveira SC, Giambartolomei GH, Cassataro JC. 2011. Confronting the barriers to develop vaccines against brucellosis. Expert. Rev. Vaccines 10: 1291-1305.
- Godfroid J, Käsbohrer A. 2002. Brucellosis in the European Union and Norway at the turn of the twenty-first century. Vet. Microbiol. 90: 135-145.
- Perkins SD, Smither SJ, Atkins HS. 2010. Towards a Brucella vaccine for humans. FEMS. Microbiol. Rev. 34: 379-394.
- Fugier E, Pappas G, Gorvel JP. 2007. Virulence factors in brucellosis: implications for aetiopathogenesis and treatment. Expert. Rev. Mol. Med. 9: 1-10.
- Jiang X, Baldwin CL. 1993. Iron augments macrophagemediated killing of Brucella abortus alone and in conjunction with IFN-γ. Cell. Immunol. 148: 397-407.
- Luo D, Ni B, Li P, Shi W, Zhang S, Han Y. 2006. Protective immunity elicited by a divalent DNA vaccine encoding both the L7/L12 and Omp16 genes of Brucella abortus in BALB/c mice. Infect. Immun. 74: 2734-2741.
- Motaharinia Y, Rezaee MA, Rashidi A, Jalili A, Rezaie MJ, Shapouri R, et al. 2013. Induction of protective immunity against brucellosis in mice by vaccination with a combination of naloxone, alum and heat-killed Brucella melitensis 16M. J. Microbiol. Immunol. Infect. 46: 253-258.
- Cassataro J, Estein SM, Pasquevich KA, Velikovsky CA, de la Barrera S, Bowden R, et al. 2005. Vaccination with the recombinant Brucella outer membrane protein 31 or a derived 27-amino-acid synthetic peptide elicits a CD4+ T helper 1 response that protects against Brucella melitensis infection. Infect. Immun. 73: 8079-8088.
- Pasquevich KA, Estein SM, Garcia-Samartino C, Zwerdling A, Coria LM, Barrionuevo P, et al. 2009. Immunization with recombinant Brucella species outer membrane protein Omp16 and Omp19 in adjuvant induces specific CD4+ and CD8+ T cells as well as systemic and oral protection against Brucella abortus infection. Infect. Immun. 77: 436-445.
- Cou per KN, Blount DG, R iley EM. 2 008. IL-10; the m aster regulator of immunity to infection. J. Immunol. 180: 5771-5777.
- Fernandez D M, Baldwin C L. 1 995. I nterleu ink-10 downregulates protective immunity to Brucella abortus. Infect. Immun. 63: 1130-1133.
- Corsetti P, de Almeida L, Carvalho N, Azevedo V, Teane S, Texeira H, et al. 2013. Lack of endogenous IL-10 enhances production of proinflammatory cytokines and leads to Brucella abortus clearance in mice. PLoS One 8: e74729.
- Hop HT, Arayan LT, Tran Xuan H, Reyes AW, Kim S. 2018. Immunization of BALB/c mice with a combination of four recombinant Brucella abortus proteins, AspC, Dps, InpB and Ndk, confers a marked protection against a virulent strain of Brucella abortus. Vaccine 36: 3027-3033.