Abstract
Brucellosis is the most common zoonotic bacterial disease. Prevention of human brucellosis is achieved through pasteurization of dairy products, appropriate sanitation and vaccination of domestic animals against the Brucella species. B. abortus unlipidated 19 kDa outer membrane protein (U-Omp19) is a promising candidate for a subunit vaccine against brucellosis. This study investigates immunogenicity of Omp19 alone and with Freund’s adjuvant (Omp19-IFA) and N-trimethyl chitosan (TMC/Omp19) nanoparticles, as well as the effect of Omp19 administration route on immunological responses and protection. The omp19 gene was expressed in E. coli BL21 (DE3). After purification, the recombinant Omp19 was loaded onto TMC nanoparticles by ionic gelation with tripolyphosphate. Particle size and loading efficiency of the nanoparticles were determined. Omp19-IFA was administered intraperitoneally while TMC/Omp19 nanoparticles were administered orally and intraperitoneally. The results indicated that intraperitoneal (i.p.) immunization by Omp19-IFA and TMC/Omp19 nanoparticles induced Th1 and Th2 immune responses, respectively, whereas oral immunization of TMC/Omp19 nanoparticles induced a mixed Th1/Th17 immune response. Moreover, oral immunization increased IgA levels in feces. Immunized mice were challenged with virulent B. melitensis 16 M and B. abortus 544. Oral immunization with TMC/Omp19 nanoparticles induced a remarkably high protection level against B. melitensis and B. abortus. The results showed that immunization route has a pivotal role in immune response polarization and protective efficiency of Omp19 antigen. Also, it was deduced that the higher protection level achieved through oral administration of TMC/Omp19 nanoparticles may be due to the elicited Th17 response.
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References
Abkar M, Amani J, Sahebghadam Lotfi A, Nikbakht Brujeni G, Alamian S, Kamali M (2015) Subcutaneous immunization with a novel immunogenic candidate (urease) confers protection against Brucella abortus and Brucella melitensis infections. APMIS: Acta Pathol Microbiol Immunol Scand 123:667–675. doi:10.1111/apm.12400
Amidi M et al (2007) N-trimethyl chitosan (TMC) nanoparticles loaded with influenza subunit antigen for intranasal vaccination: biological properties and immunogenicity in a mouse model. Vaccine 25:144–153. doi:10.1016/j.vaccine.2006.06.086
Ariza J, Gudiol F, Pallares R, Viladrich PF, Rufi G, Corredoira J, Miravitlles MR (1992) Treatment of human brucellosis with doxycycline plus rifampin or doxycycline plus streptomycin. A randomized, double-blind study. Ann Intern Med 117:25–30
Bal SM, Slutter B, Jiskoot W, Bouwstra JA (2011) Small is beautiful: N-trimethyl chitosan-ovalbumin conjugates for microneedle-based transcutaneous immunisation. Vaccine 29:4025–4032. doi:10.1016/j.vaccine.2011.03.039
Baumann U (2008) Mucosal vaccination against bacterial respiratory infections. Expert Rev Vaccines 7:1257–1276
Brown DM, Kamperschroer C, Dilzer AM, Roberts DM, Swain SL (2009) IL-2 and antigen dose differentially regulate perforin- and FasL-mediated cytolytic activity in antigen specific CD4+ T cells. Cell Immunol 257:69–79. doi:10.1016/j.cellimm.2009.03.002
Calvo P, Remunan-Lopez C, Vila-Jato JL, Alonso MJ (1997) Chitosan and chitosan/ethylene oxide-propylene oxide block copolymer nanoparticles as novel carriers for proteins and vaccines. Pharm Res 14:1431–1436
Cassataro J 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
Chen F, Zhang ZR, Yuan F, Qin X, Wang M, Huang Y (2008) In vitro and in vivo study of N-trimethyl chitosan nanoparticles for oral protein delivery. Int J Pharm 349:226–233. doi:10.1016/j.ijpharm.2007.07.035
Clapp B, Skyberg JA, Yang X, Thornburg T, Walters N, Pascual DW (2011) Protective live oral brucellosis vaccines stimulate Th1 and th17 cell responses. Infect Immun 79:4165–4174. doi:10.1128/IAI.05080-11
Cloeckaert A, Verger JM, Grayon M, Paquet JY, Garin-Bastuji B, Foster G, Godfroid J (2001) Classification of Brucella spp. isolated from marine mammals by DNA polymorphism at the omp2 locus. Microbes Infect 3:729–738
Commander NJ, Spencer SA, Wren BW, MacMillan AP (2007) The identification of two protective DNA vaccines from a panel of five plasmid constructs encoding Brucella melitensis 16M genes. Vaccine 25:43–54. doi:10.1016/j.vaccine.2006.07.046
Daniel LW, Huang C, Strum JC, Smitherman PK, Greene D, Wykle RL (1993) Phospholipase D hydrolysis of choline phosphoglycerides is selective for the alkyl-linked subclass of Madin-Darby canine kidney cells. J Biol Chem 268:21519–21526
Fiorentino MA, Campos E, Cravero S, Arese A, Paolicchi F, Campero C, Rossetti O (2008) Protection levels in vaccinated heifers with experimental vaccines Brucella abortus M1-luc and INTA 2. Vet Microbiol 132:302–311. doi:10.1016/j.vetmic.2008.05.003
Fooks AR (2000) Development of oral vaccines for human use. Curr Opin Mol Ther 2:80
Fu S et al (2012) Immunization of mice with recombinant protein CobB or AsnC confers protection against Brucella abortus infection. PLoS One 7:e29552. doi:10.1371/journal.pone.0029552
Garg NK, Mangal S, Khambete H, Tyagi RK (2010) Mucosal delivery of vaccines: role of mucoadhesive/biodegradable polymers. Recent Pat Drug Deliv Formul 4:114–128
Gebert A, Steinmetz I, Fassbender S, Wendlandt KH (2004) Antigen transport into Peyer’s patches: increased uptake by constant numbers of M cells. Am J Pathol 164:65–72. doi:10.1016/S0002-9440(10)63097-0
Ghasemi A, Jeddi-Tehrani M, Mautner J, Salari MH, Zarnani AH (2014a) Immunization of mice with a novel recombinant molecular chaperon confers protection against Brucella melitensis infection. Vaccine. doi:10.1016/j.vaccine.2014.09.013
Ghasemi A, Zarnani AH, Ghoodjani A, Rezania S, Salari MH, Jeddi-Tehrani M (2014b) Identification of a new immunogenic candidate conferring protection against Brucella melitensis infection in Mice. Mol Immunol 62:142–149. doi:10.1016/j.molimm.2014.06.017
Giambartolomei GH, Zwerdling A, Cassataro J, Bruno L, Fossati CA, Philipp MT (2004) Lipoproteins, not lipopolysaccharide, are the key mediators of the proinflammatory response elicited by heat-killed Brucella abortus. J Immunol 173:4635–4642
Goel D, Bhatnagar R (2012) Intradermal immunization with outer membrane protein 25 protects Balb/c mice from virulent B. abortus 544. Mol Immunol 51:159–168. doi:10.1016/j.molimm.2012.02.126
Goel D, Rajendran V, Ghosh PC, Bhatnagar R (2013) Cell mediated immune response after challenge in Omp25 liposome immunized mice contributes to protection against virulent Brucella abortus 544. Vaccine 31:1231–1237. doi:10.1016/j.vaccine.2012.12.043
Golding B et al (2001) Immunity and protection against Brucella abortus. Microbes Infect 3:43–48
Gregory AE, Titball R, Williamson D (2013) Vaccine delivery using nanoparticles. Front Cell Infect Microbiol 3:13. doi:10.3389/fcimb.2013.00013
Hall JB, Dobrovolskaia MA, Patri AK, McNeil SE (2007) Characterization of nanoparticles for therapeutics. Nanomedicine (Lond) 2:789–803. doi:10.2217/17435889.2.6.789
Jabbal-Gill I, Watts P, Smith A (2012) Chitosan-based delivery systems for mucosal vaccines. Expert Opin Drug Deliv 9:1051–1067. doi:10.1517/17425247.2012.697455
Johansen P, Mohanan D, Martinez-Gomez JM, Kundig TM, Gander B (2010) Lympho-geographical concepts in vaccine delivery. J Control Release 148:56–62. doi:10.1016/j.jconrel.2010.05.019
Kammona O, Kiparissides C (2012) Recent advances in nanocarrier-based mucosal delivery of biomolecules. J Control Release 161:781–794. doi:10.1016/j.jconrel.2012.05.040
Kostrzak A et al (2009) Oral administration of low doses of plant-based HBsAg induced antigen-specific IgAs and IgGs in mice, without increasing levels of regulatory T cells. Vaccine 27:4798–4807
Kumar P, Chen K, Kolls JK (2013) Th17 cell based vaccines in mucosal immunity. Curr Opin Immunol 25:373–380. doi:10.1016/j.coi.2013.03.011
Luna-Martinez JE, Mejia-Teran C (2002) Brucellosis in Mexico: current status and trends. Vet Microbiol 90:19–30
Luo D et al (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. doi:10.1128/IAI.74.5.2734-2741.2006
Mahapatro A, Singh DK (2011) Biodegradable nanoparticles are excellent vehicle for site directed in-vivo delivery of drugs and vaccines. J Nanobiotechnol 9:55. doi:10.1186/1477-3155-9-55
Mobley HL, Island MD, Hausinger RP (1995) Molecular biology of microbial ureases. Microbiol Rev 59:451–480
Mohanan D et al (2010) Administration routes affect the quality of immune responses: a cross-sectional evaluation of particulate antigen-delivery systems. J Control Release 147:342–349. doi:10.1016/j.jconrel.2010.08.012
Montejo JM, Alberola I, Glez-Zarate P, Alvarez A, Alonso J, Canovas A, Aguirre C (1993) Open, randomized therapeutic trial of six antimicrobial regimens in the treatment of human brucellosis. Clin Infect Dis 16:671–676
Murphy EA, Sathiyaseelan J, Parent MA, Zou B, Baldwin CL (2001) Interferon-gamma is crucial for surviving a Brucella abortus infection in both resistant C57BL/6 and susceptible BALB/c mice. Immunology 103:511–518
Neutra MR, Kozlowski PA (2006) Mucosal vaccines: the promise and the challenge. Nat Rev Immunol 6:148–158
Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV (2006) The new global map of human brucellosis. Lancet Infect Dis 6:91–99. doi:10.1016/S1473-3099(06)70382-6
Pasquevich KA et al (2009) Immunization with recombinant Brucella species outer membrane protein Omp16 or 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. doi:10.1128/IAI.01151-08
Pasquevich KA et al (2011) An oral vaccine based on U-Omp19 induces protection against B. abortus mucosal challenge by inducing an adaptive IL-17 immune response in mice. PLoS One 6:e16203
Perkins SD, Smither SJ, Atkins HS (2010) Towards a Brucella vaccine for humans. FEMS Microbiol Rev 34:379–394
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. doi:10.1016/j.ijpharm.2008.04.036
Saraiva M, Christensen JR, Veldhoen M, Murphy TL, Murphy KM, O’Garra A (2009) Interleukin-10 production by Th1 cells requires interleukin-12-induced STAT4 transcription factor and ERK MAP kinase activation by high antigen dose. Immunity 31:209–219. doi:10.1016/j.immuni.2009.05.012
Shakir RA (1986) Neurobrucellosis. Postgrad Med J 62:1077–1079
Slutter B et al (2009) Mechanistic study of the adjuvant effect of biodegradable nanoparticles in mucosal vaccination. J Control Release 138:113–121. doi:10.1016/j.jconrel.2009.05.011
Subbiah R, Ramalingam P, Ramasundaram S, Kim do Y, Park K, Ramasamy MK, Choi KJ (2012) N,N,N-Trimethyl chitosan nanoparticles for controlled intranasal delivery of HBV surface antigen. Carbohydr Polym 89:1289–1297. doi:10.1016/j.carbpol.2012.04.056
Tabynov K et al (2014a) Influenza viral vectors expressing the Brucella OMP16 or L7/L12 proteins as vaccines against B. abortus infection. Virol J 11:69. doi:10.1186/1743-422X-11-69
Tabynov K, Yespembetov B, Sansyzbay A (2014b) Novel vector vaccine against Brucella abortus based on influenza A viruses expressing Brucella L7/L12 or Omp16 proteins: evaluation of protection in pregnant heifers. Vaccine 32:5889–5892. doi:10.1016/j.vaccine.2014.08.073
Vemulapalli R et al (2000) Characterization of specific immune responses of mice inoculated with recombinant vaccinia virus expressing an 18-kilodalton outer membrane protein of Brucella abortus. Clin Diagn Lab Immunol 7:114–118
Verheul RJ, Slutter B, Bal SM, Bouwstra JA, Jiskoot W, Hennink WE (2011) Covalently stabilized trimethyl chitosan-hyaluronic acid nanoparticles for nasal and intradermal vaccination. J Control Release 156:46–52. doi:10.1016/j.jconrel.2011.07.014
Vitry MA et al (2012) Crucial role of gamma interferon-producing CD4+ Th1 cells but dispensable function of CD8+ T cell, B cell, Th2, and Th17 responses in the control of Brucella melitensis infection in mice. Infect Immun 80:4271–4280. doi:10.1128/IAI.00761-12
Acknowledgments
This work was supported by INSF (No: 91000787): Iran National Science Foundation. We would like to thank Dr. Bram Slutter and Dr. Nima Khoramabadi for their helpful advice.
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The authors declare that they have no conflicts of interest.
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Abkar, M., Lotfi, A.S., Amani, J. et al. Survey of Omp19 immunogenicity against Brucella abortus and Brucella melitensis: influence of nanoparticulation versus traditional immunization. Vet Res Commun 39, 217–228 (2015). https://doi.org/10.1007/s11259-015-9645-2
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DOI: https://doi.org/10.1007/s11259-015-9645-2