Evaluation of a DNA vaccine encoding Brucella BvrR in BALB/c mice

Chen,Bo; Liu,Baoshan; Zhao,Zhina; Wang,Guizhen

doi:10.3892/mmr.2018.9735

Evaluation of a DNA vaccine encoding Brucella BvrR in BALB/c mice

Authors:
- Bo Chen
- Baoshan Liu
- Zhina Zhao
- Guizhen Wang
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Affiliations: Department of Pathogenic Biology, College of Basic Medical Science, China Medical University, Shenyang, Liaoning 110122, P.R. China, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, Liaoning 110866, P.R. China, Department of Microbiology and Cell Biology, College of Life Science and Pharmaceuticals, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, P.R. China
Published online on: December 11, 2018 https://doi.org/10.3892/mmr.2018.9735
Pages: 1302-1308

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Abstract

Brucellosis is an important neglected zoonotic disease, and the pathogens responsible are Brucellae. In order to evaluate the immunogenicity and protective efficacy of a DNA vaccine encoding Brucella BvrR, the recombinant plasmid pCDNA‑BvrR was constructed by inserting the BvrR gene fragment into a pCDNA3.0 vector. The His6‑tagged BvrR was purified with His‑trap FF crude affinity chromatography and verified with an anti‑histidine monoclonal antibody by western blot analysis. The specific immunoglobulin antigens and their isotypes were detected by indirect ELISA. The recombinant His6‑BvrR protein was expressed and purified by affinity chromatography. The optical density 450 value of immunoglobulin G (IgG) in the pCDNA‑BvrR group was significantly increased compared with the pCDNA3.0 vector or PBS groups (P<0.05), and the pCDNA3.0 vector and PBS groups exhibited no significant difference (P>0.05). BvrR induced specific antibodies with a dominance of IgG2a over IgG1 and the T cell‑proliferative response, in addition to a typical T helper‑1 (Th1)‑dominated immune response in mice. The splenocytes from mice of the pCDNA‑BvrR group demonstrated significant proliferative activity compared with the pCDNA3.0 vector group. The present results indicated that immunization with BvrR induced a specific Th1‑type immune response in mice. Subsequent to challenging with B. abortus S19, it was identified that the DNA vaccine pCDNA‑BvrR induced a significant level of protection in BALB/c mice by evaluating systemic bacterial clearance. These results suggested that BvrR may be a good candidate for a DNA vaccine against brucellosis.

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1	Norman FF, Monge-Maillo B, Chamorro-Tojeiro S, Pérez-Molina JA and López-Vélez R: Imported brucellosis: A case series and literature review. Travel Med Infect Dis. 14:182–199. 2016. View Article : Google Scholar : PubMed/NCBI
2	Adone R, Muscillo M, La Rosa G, Francia M and Tarantino M: Antigenic, immunologic and genetic characterization of rough strains B. abortus RB51, B. melitensis B115 and B. melitensis B18. PLoS One. 6:e240732011. View Article : Google Scholar : PubMed/NCBI
3	Davis DS and Elzer PH: Brucella vaccines in wildlife. Vet Microbiol. 90:533–544. 2002. View Article : Google Scholar : PubMed/NCBI
4	Perkins SD, Smither SJ and Atkins HS: Towards a Brucella vaccine for humans. FEMS Microbiol Rev. 34:379–394. 2010. View Article : Google Scholar : PubMed/NCBI
5	Bobbala S and Hook S: Is there an optimal formulation and delivery strategy for subunit vaccines? Pharm Res. 33:2078–2097. 2016. View Article : Google Scholar : PubMed/NCBI
6	Baloglu S, Boyle SM, Vemulapalli R, Sriranganathan N, Schurig GG and Toth TE: Immune responses of mice to vaccinia virus recombinants expressing either Listeria monocytogenes partial listeriolysin or Brucella abortus ribosomal L7/L12 protein. Vet Microbiol. 109:11–17. 2005. View Article : Google Scholar : PubMed/NCBI
7	Vemulapalli R, He Y, Cravero S, Sriranganathan N, Boyle SM and Schurig GG: Overexpression of protective antigen as a novel approach to enhance vaccine efficacy of Brucella abortus strain RB51. Infect Immun. 68:3286–3289. 2000. View Article : Google Scholar : PubMed/NCBI
8	Rajasekaran P, Seleem MN, Contreras A, Purwantini E, Schurig GG, Sriranganathan N and Boyle SM: Brucella abortus strain RB51 leucine auxotroph as an environmentally safe vaccine for plasmid maintenance and antigen overexpression. Appl Environ Microbiol. 74:7051–7055. 2008. View Article : Google Scholar : PubMed/NCBI
9	Yu DH, Hu XD and Cai H: A combined DNA vaccine encoding BCSP31, SOD, and L7/L12 confers high protection against Brucella abortus 2308 by inducing specific CTL responses. DNA Cell Biol. 26:435–443. 2007. View Article : Google Scholar : PubMed/NCBI
10	Hu XD, Yu DH, Chen ST, Li SX and Hong C: A combined DNA vaccine provides protective immunity against Mycobacterium bovis Brucella abortus in cattle. DNA Cell Biol. 28:191–199. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zhao HP, Sun JF, Li N, Sun Y, Xia ZH, Wang Y, Cheng D, Qi QF, Jin ML and Qiu HJ: Assessment of the cell-mediated immunity induced by alphavirus replicon-vectored DNA vaccines against classical swine fever in a mouse model. Vet Immunol Immunopathol. 129:57–65. 2009. View Article : Google Scholar : PubMed/NCBI
12	Maspi N, Ghaffarifar F, Sharifi Z and Dalimi A: Codelivery of DNA vaccination encoding LeIF gene and IL-12 increases protection against Leishmania major infection in BALB/c mice. Parasite Immunol. 38:228–235. 2016. View Article : Google Scholar : PubMed/NCBI
13	Oksanen KE, Myllymäki H, Ahava MJ, Mäkinen L, Parikka M and Rämet M: DNA vaccination boosts Bacillus Calmette-Guérin protection against mycobacterial infection in zebrafish. Dev Comp Immunol. 54:89–96. 2016. View Article : Google Scholar : PubMed/NCBI
14	Griffin BD, Muthumani K, Warner BM, Majer A, Hagan M, Audet J, Stein DR, Ranadheera C, Racine T, De La Vega MA, et al: DNA vaccination protects mice against Zika virus-induced damage to the testes. Nat Commun. 8:157432017. View Article : Google Scholar : PubMed/NCBI
15	Bello J, Sáez D, Escalona E, Velozo P, Santiviago CA, Contreras I and Oñate Á: Mucosal immunization of BALB/c mice with DNA vaccines encoding the SEN1002 and SEN1395 open reading frames of Salmonella enterica serovar Enteritidis induces protective immunity. Epidemiol Infect. 144:247–256. 2016. View Article : Google Scholar : PubMed/NCBI
16	González-Smith A, Vemulapalli R, Andrews E and Oñate A: Evaluation of Brucella abortus DNA vaccine by expression of Cu-Zn superoxide dismutase antigen fused to IL-2. Immunobiology. 211:65–74. 2006. View Article : Google Scholar : PubMed/NCBI
17	Velikovsky CA, Cassataro J, Giambartolomei GH, Goldbaum FA, Estein S, Bowden RA, Bruno L, Fossati CA and Spitz M: A DNA vaccine encoding lumazine synthase from Brucella abortus induces protective immunity in BALB/c mice. Infect Immun. 70:2507–2511. 2002. View Article : Google Scholar : PubMed/NCBI
18	López-Goñi I, Guzmán-Verri C, Manterola L, Sola-Landa A, Moriyón I and Moreno E: Regulation of Brucella virulence by the two-component system BvrR/BvrS. Vet Microbiol. 90:329–339. 2002. View Article : Google Scholar : PubMed/NCBI
19	Guzmán-Verri C, Manterola L, Sola-Landa A, Parra A, Cloeckaert A, Garin J, Gorvel JP, Moriyon I, Moreno E and Lopez-Goni I: The two-component system BvrR/BvrS essential for Brucella abortus virulence regulates the expression of outer membrane proteins with counterparts in members of the Rhizobiaceae. Proc Natl Acad Sci USA. 99:12375–12380. 2002. View Article : Google Scholar : PubMed/NCBI
20	Manterola L, Moriyón I, Moreno E, Sola-Landa A, Weiss DS, Koch MH, Howe J, Brandenburg K and López-Goñi I: The lipopolysaccharide of Brucella abortus BvrS/BvrR mutants contains lipid a modifications and has higher affinity for bactericidal cationic peptides. J Bacteriol. 187:5631–5639. 2005. View Article : Google Scholar : PubMed/NCBI
21	Manterola L, Guzmán-Verri C, Chaves-Olarte E, Barquero-Calvo E, de Miguel MJ, Moriyón I, Grilló MJ, López-Goñi I and Moreno E: BvrR/BvrS-controlled outer membrane proteins Omp3a and Omp3b are not essential for Brucella abortus virulence. Infect Immun. 75:4867–4874. 2007. View Article : Google Scholar : PubMed/NCBI
22	Martínez-Núñez C, Altamirano-Silva P, Alvarado-Guillén F, Moreno E, Guzmán-Verri C and Chaves-Olarte E: The two-component system BvrR/BvrS regulates the expression of the type IV secretion system VirB in Brucella abortus. J Bacteriol. 192:5603–5608. 2010. View Article : Google Scholar : PubMed/NCBI
23	Tadesse G: Brucellosis seropositivity in animals and humans in ethiopia: A meta-analysis. PLoS Negl Trop Dis. 10:e00050062016. View Article : Google Scholar : PubMed/NCBI
24	Faurez F, Grasland B, Béven V, Cariolet R, Keranflec'h A, Henry A, Jestin A and Dory D: The protective immune response against Pseudorabies virus induced by DNA vaccination is impaired if the plasmid harbors a functional Porcine circovirus type 2 rep and origin of replication. Antiviral Res. 96:271–279. 2012. View Article : Google Scholar : PubMed/NCBI
25	Otani N, Yamanishi K, Sakaguchi Y, Imai Y, Shima M and Okuno T: Varicella-zoster virus-specific cell-mediated immunity in subjects with herpes zoster. J Immunol Methods. 377:53–55. 2012. View Article : Google Scholar : PubMed/NCBI
26	Tan Z, Zhou T, Zheng H, Ding Y and Xu Y: Malaria DNA vaccine gp96NTD-CSP elicits both CSP-specific antibody and CD8(+) T cell response. Parasitol Res. 114:2333–2339. 2015. View Article : Google Scholar : PubMed/NCBI
27	Estein SM, Cheves PC, Fiorentino MA, Cassataro J, Paolicchi FA and Bowden RA: Immunogenicity of recombinant Omp31 from Brucella melitensis in rams and serum bactericidal activity against B. ovis. Vet Microbiol. 102:203–213. 2004. View Article : Google Scholar : PubMed/NCBI
28	Lacerda TL, Salcedo SP and Gorvel JP: Brucella T4SS: The VIP pass inside host cells. Curr Opin Microbiol. 16:45–51. 2013. View Article : Google Scholar : PubMed/NCBI
29	Oñate AA, Céspedes S, Cabrera A, Rivers R, González A, Muñoz C, Folch H and Andrews E: A DNA vaccine encoding Cu, Zn superoxide dismutase of Brucella abortus induces protective immunity in BALB/c mice. Infect Immun. 71:4857–4861. 2003. View Article : Google Scholar : PubMed/NCBI
30	Avila-Calderón ED, Lopez-Merino A, Sriranganathan N, Boyle SM and Contreras-Rodríguez A: A history of the development of Brucella vaccines. Biomed Res Int. 2013:7435092013. View Article : Google Scholar : PubMed/NCBI
31	Ranjbar R, Sharifimoghadam S, Saeidi E, Jonaidi N and Sheikhshahrokh A: Induction of protective immune responses in mice by double DNA vaccine encoding of Brucella melitensis Omp31 and Escherichia coli Eae genes. Trop J Pharm Res. 15:20772016. View Article : Google Scholar
32	Yu DH, Li M, Hu XD and Cai H: A combined DNA vaccine enhances protective immunity against Mycobacterium tuberculosis Brucella abortus in the presence of an IL-12 expression vector. Vaccine. 25:6744–6754. 2007. View Article : Google Scholar : PubMed/NCBI
33	Lladser A, Párraga M, Quevedo L, Carmen Molina M, Silva S, Ferreira A, Billetta R and G Quest AF: Naked DNA immunization as an approach to target the generic tumor antigen survivin induces humoral and cellular immune responses in mice. Immunobiology. 211:11–27. 2006. View Article : Google Scholar : PubMed/NCBI
34	Hinkula J, Devignot S, Åkerström S, Karlberg H, Wattrang E, Bereczky S, Mousavi-Jazi M, Risinger C, Lindegren G, Vernersson C, et al: Immunization with DNA plasmids coding for crimean-congo hemorrhagic fever virus capsid and envelope proteins and/or virus-like particles induces protection and survival in challenged mice. J Virol. 91:e02076–e02116. 2017. View Article : Google Scholar : PubMed/NCBI