The combined use of recombinant and pVAX-omp31 DNA Vaccine for immunological protection against pathogenic Brucellas melitesnsis in an experimental model of BALB/c Mice

Harzandi, Naser; Aghababa, Haniyeh; Khoramabadi, Nima; Tabaraie, Bahman

year 11, Issue 1 (March - April 2017) Iran J Med Microbiol 2017, 11(1): 39-47 | Back to browse issues page

Mendeley

Zotero

RefWorks

Harzandi N, Aghababa H, Khoramabadi N, Tabaraie B. The combined use of recombinant and pVAX-omp31 DNA Vaccine for immunological protection against pathogenic Brucellas melitesnsis in an experimental model of BALB/c Mice . Iran J Med Microbiol 2017; 11 (1) :39-47
URL: http://ijmm.ir/article-1-566-en.html

The combined use of recombinant and pVAX-omp31 DNA Vaccine for immunological protection against pathogenic Brucellas melitesnsis in an experimental model of BALB/c Mice

Naser Harzandi

¹, Haniyeh Aghababa²

, Nima Khoramabadi²

, Bahman Tabaraie³

1- Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran , naser.harzandi@kiau.ac.ir
2- Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
3- Pasteur Institute of Iran, Tehran, Iran

Abstract: (9268 Views)

Background and Aim: Brucellosis is still an important zoonotic infection and evaluation of immunologic properties of various bacterial antigens along with different vaccination strategies helps in designing efficient vaccines against the disease. The aim of this study is to immunological evaluate the eukaryotic vector pVAX1, carrying the outer membrane protein gene of 31 kDa (Omp31) B.melitensis.

Materials and Methods: In this study which was carried out in 2014, whole sequence of omp31 of B. melitensis was inserted between BamHI and XhoI of pVAX1 plasmid vector. Female BALB/c mice aged 6-8 weeks (purchased from Pasteur Institute of Iran) were immunized intra-muscularly with 100μg of the construct, followed by either protein or plasmid boosters separately. The level of IL-4, IL-12, IFN-γ, total serum IgG, and specific IgG1 and IgG2a against recombinant Omp31 were evaluated. Finally the protective immune response following exposure to B. melitensis 16M was evaluated.

Results: DNA-vaccine omp31 career with protein reminders Omp31, stimulate higher levels of IFN-γ, IL-12 and IgG2a compared to groups of DNA-vaccine or recombinant protein. Protective immunity was also significantly higher in mice which immunized with DNA vaccine– protein regimen.

Conclusions: Mice which immunized with DNA vaccine–protein regimen showed a significantly higher levels of IL-12 and IFN-γ along with serum IgG2a which together imply augmentation of T cell-mediated immune responses against Omp31. The latter was confirmed by significant protective response to B. melitensis 16M challenge.

Keywords: B. melitensis, DNA vaccine, Omp31, pVAX1

Full-Text [PDF 814 kb] (2346 Downloads)

Type of Study: Original Research Article | Subject: Molecular Microbiology
Received: 2016/05/28 | Accepted: 2017/02/15 | ePublished: 2017/03/16

References

1. von Bargen K, Gorvel JP, Salcedo SP. Internal affairs: investigating the Brucella intracellular lifestyle. FEMS Microbiol Rev 2012; 36(3):533-62. [PubMed]

2. Ramin B, Macpherson P. Human brucellosis. Bmj 2010; 341:c4545.

3. Mitka S, Ifantidou A, Chaidouli E, Vavatsi N, Kansouzidou A. Human brucellosis from Rev. 1 vaccine strain. Differentiation of the field Brucella strains from Rev. 1 vaccine strain. Acta Microbiologica Hellenica 2005; 50(6):352-64. [Article]

4. Schurig GG, Sriranganathan N, Corbel MJ. Brucellosis vaccines: past, present and future. Vet Microbiol 2002; 90(1-4):479-96. [PubMed]

5. Martirosyan A, Moreno E, Gorvel JP. An evolutionary strategy for a stealthy intracellular Brucella pathogen. Immunol Rev 2011; 240(1):211-34. [PubMed]

6. Celli J. Surviving inside a macrophage: the many ways of Brucella. Res Microbiol 2006; 157(2):93-8. [Article]

7. Martirosyan A, Gorvel JP. Brucella evasion of adaptive immunity. Fut Microbiol 2013; 8(2):147-54. [PubMed]

8. Dornand J, Gross A, Lafont V, Liautard J, Oliaro J, Liautard JP. The innate immune response against Brucella in humans. Vet Microbiol 2002; 90(1-4):383-94. [PubMed]

9. Ko J, Splitter GA. Molecular host-pathogen interaction in brucellosis: current understanding and future approaches to vaccine development for mice and humans. Clin Microbiol Rev 2003; 16(1):65-78. [PubMed]

10. Sung KY, Jung M, Shin MK, Park HE, Lee JJ, Kim S, et al. 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 2014; 24(6):854-61. [Article]

11. Ibanez AE, Smaldini P, Coria LM, Delpino MV, Pacifico LG, Oliveira SC, Risso GS, Pasquevich KA, Fossati CA, Giambartolomei GH, Docena GH, Cassataro J. Unlipidated Outer Membrane Protein Omp16 (U-Omp16) from Brucella spp. as Nasal Adjuvant Induces a Th1 Immune Response and Modulates the Th2 Allergic Response to Cow's Milk Proteins. PloS one 2013; 8(7):e69438. [Article]

12. Goel D, Bhatnagar R. Intradermal immunization with outer membrane protein 25 protects Balb/c mice from virulent B. abortus 544. Mol Immunol 2012; 51(2):159-68. [PubMed]

13. Kaushik P, Singh DK. Cytokine profile of mice immunized with 28kDa outer membrane protein of brucella melitensis. Indian Vet J 2011; 88(8):15-7.

14. Jain S, Afley P, Dohre SK, Saxena N, Kumar S. Evaluation of immunogenicity and protective efficacy of a plasmid DNA vaccine encoding ribosomal protein L9 of Brucella abortus in BALB/c mice. Vaccine 2014; 32(35):4537-42. [PubMed]

15. Riquelme-Neira R, Retamal-Diaz A, Acuna F, Riquelme P, Rivera A, Saez D, Onate A. Protective effect of a DNA vaccine containing an open reading frame with homology to an ABC-type transporter present in the genomic island 3 of Brucella abortus in BALB/c mice. Vaccine 2013; 31(36):3663-7. [PubMed]

16. Sislema-Egas F, Cespedes S, Fernandez P, Retamal-Diaz A, Saez D, Onate A. Evaluation of protective effect of DNA vaccines encoding the BAB1_0263 and BAB1_0278 open reading frames of Brucella abortus in BALB/c mice. Vaccine 2012; 30(50):7286-91. [Article]

17. Doosti A, Ghasemi-Dehkordi P, Javadi GR, Sardari S, Shokrgozar MA. DNA vaccine encoding the Omp31 gene of Brucella melitensis induces protective immunity in BALB/c mice. Res J Bio Sci 2009; 4(1):126-31. [Article]

18. Gupta VK, Rout PK, Vihan VS. Induction of immune response in mice with a DNA vaccine encoding outer membrane protein (omp31) of Brucella melitensis 16M. Res Vet Sci 2007; 82(3):305-13. [Article]

19. Delpino MV, Cassataro J, Fossati CA, Goldbaum FA, Baldi PC. Brucella outer membrane protein Omp31 is a haemin-binding protein. Microbes Infect 2006; 8(5):1203-8. [Article]

20. Cassataro J, Velikovsky CA, Bruno L, Estein SM, de la Barrera S, Bowden R, Fossati CA, Giambartolomei GH. Improved immunogenicity of a vaccination regimen combining a DNA vaccine encoding Brucella melitensis outer membrane protein 31 (Omp31) and recombinant Omp31 boosting. Clin Vaccine Immunol 2007; 14(7):869-74. [PubMed]

21. Cassataro J, Pasquevich KA, Estein SM, Laplagne DA, Zwerdling A, de la Barrera S, et al. A DNA vaccine coding for the chimera BLSOmp31 induced a better degree of protection against B. ovis and a similar degree of protection against B. melitensis than Rev.1 vaccination. Vaccine 2007; 25(32):5958-67.

22. Estein SM, Cheves PC, Fiorentino MA, Cassataro J, Paolicchi FA, Bowden RA. Immunogenicity of recombinant Omp31 from Brucella melitensis in rams and serum bactericidal activity against B. ovis. Vet Microbiol 2004; 102(3-4):203-13. [PubMed]

23. Khodadadi S, Mohabati Mobarez A, Harzandi N, Tabaraei B, Khoramabadi N, Bakhtiyari A, et al. Cloning and expression of 31 KDa outer membrane protein of Brucella melitensis in E. coli. J Ardabil Univ Med Sci 2012;12(1):33-39 [Article]

24. Aghababa H, Mohabati Mobarez A, Khoramabadi N, Behmanesh M, Mahdavi M, Tebianian M, et al. A comparative approach to strategies for cloning, expression, and purification of Mycobacterium tuberculosis mycolyl transferase 85B and evaluation of immune responses in BALB/c mice. Mol Biotechnol 2014; 56(6):487-97. [PubMed]

25. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acid Res 1987; 15(20):8125-48. [PubMed]

26. Cassataro J, Pasquevich K, Bruno L, Wallach JC, Fossati CA, Baldi PC. Antibody Reactivity to Omp31 from Brucella melitensis in Human and Animal Infections by Smooth and Rough Brucellae. Clin Diagn Lab Immunol 2004; 11(1):111-4. [PubMed]

27. Al-Mariri A, Abbady AQ. Evaluation of the immunogenicity and the protective efficacy in mice of a DNA vaccine encoding SP41 from Brucella melitensis. J Infect dev Ctries 2013; 7(4):329-37.

28. Yu DH, Hu XD, 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 2007; 26(6):435-43. [PubMed]

29. Cassataro J, Velikovsky CA, de la Barrera S, Estein SM, Bruno L, Bowden R, et al. A DNA vaccine coding for the Brucella outer membrane protein 31 confers protection against B. melitensis and B. ovis infection by eliciting a specific cytotoxic response. Infect Immun 2005; 73(10):6537-46. [PubMed]

30. Singha H, Mallick AI, Jana C, Isore DP, Goswami TK, Srivastava SK, et al. Escheriosomes entrapped DNA vaccine co-expressing Cu-Zn superoxide dismutase and IL-18 confers protection against Brucella abortus. Microbes Infect 2008; 10(10-11):1089-96. [PubMed]

31. Luo D, Ni B, Li P, Shi W, Zhang S, Han Y, et al. 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 2006; 74(5):2734-41. [PubMed]

32. Saez D, Guzman I, Andrews E, Cabrera A, Onate A. Evaluation of Brucella abortus DNA and RNA vaccines expressing Cu-Zn superoxide dismutase (SOD) gene in cattle. Vet Microbiol 2008; 129(3-4):396-403. [Article]

33. Imani Fooladi AA, Bagherpour G, Khoramabadi N, Fallah Mehrabadi J, Mahdavi M, Halabian R, Amin M, Izadi Mobarakeh J, Einollahi B. Cellular immunity survey against urinary tract infection using pVAX/fimH cassette with mammalian and wild type codon usage as a DNA vaccine. Clin Exp Vaccine Res 2014; 3(2):185-93. [PubMed]