Dengue virus type-3 envelope protein domain III; expression and immunogenicity

Document Type : Original Article

Authors

1 Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran 2 Department of Cellular and Molecular Biology, Islamic Azad University, Pharmaceutical Sciences Branch (IAUPS), Tehran, Iran

2 Department of Immunology, Faculty of Medical Sciences, Tehran Medical University, Tehran, Iran

3 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

4 Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s): Production of a recombinant and immunogenic antigen using dengue virus type-3 envelope protein is a key point in dengue vaccine development and diagnostic researches. The goals of this study were providing a recombinant protein from dengue virus type-3 envelope protein and evaluation of its immunogenicity in mice.
Materials and Methods: Multiple amino acid sequences of different isolates of dengue virus type-3, corresponding to the envelope protein domain III, were achieved from GenBank. Clustal V alignment tool was used to provide a consensus amino acid sequence. Nucleotide sequence of the coding gene was optimized using “Optimizer”. The origami (DE3) strain of Escherichia coli was used as the host in order to express the protein. A commercial affinity chromatography method was used to purify the recombinant protein. Immunogenicity of the recombinant protein was evaluated in mice using ELISA, MTT and cytokine assays.
Results: A consensus amino acid sequence corresponding to the most important region of dengue virus type-3 envelope protein (domain III) was provided. A high concentration (≥ 20 mg/L culture medium) of soluble recombinant antigen (EDIII3) was achieved. Immunized mice developed specific antibody responses against EDIII3 protein. The splenocytes from EDIII3-immunized mice showed a high proliferation rate in comparison with the negative control. In addition, the concentrations of two measured cytokines (IFN-γ and IL-4) were increased markedly in immunized mice.
Conclusion: The results showed that the expressed recombinant EDIII3 protein is an immunogenic antigen and can be applied to induce specific immune responses against dengue virus type-3.

Keywords

References

1. Gubler DJ. Epidemic dengue/dengue hemorrhagic fever as a public health, social and economic problem in the 21st century. Trends Microbiol 2002; 10:100–103.
2. Center for Disease Control and Prevention. Available at: http://cdc.gov/ncidod/dvbid/dengue
3. Stephenson JR. Understanding dengue pathogenesis: implications for vaccine design. Bull World Health Organ 2005: 83:308-314.
4. Tong GZ, Zhou YJ, Hao XF, Tian ZJ, An TQ, Qiu HJ. Highly pathogenic porcine reproductive and respiratory syndrome, China. Emerg Infect Dis 2007; 13:1434-1436.
5. Li DJ, Wang HM, Li L. Gene fusion of molecular adjuvant C3d to hCG enhances the anti-hCG antibody response in DNA immunization. J Reprod Immunol 2003; 60:129–141.
6. Simmons M, Murphy GS, Hayes CG. Short report: Antibody responses of mice immunized with a tetravalent dengue recombinant protein subunit vaccine. Am J Trop Med Hyg 2001; 65:159-161.
7. Hermida L, Rodriguez R, Lazo L, Bernardo L, Silva R, Zulueta A, et al. A fragment of the envelope protein from dengue-1 virus, fused in two different sites of the meningococcal P64k protein carrier, induces a functional immune response in mice. Biotechnol Appl Biochem 2004; 39:107-114.
8. Raja NU, Holman DH, Wang D, Raviprakash K, Juompan LY, Deitz SB, et al. Induction of bivalent immune responses by expression of dengue virus type 1 and type 2 antigens from a single complex adenoviral vector. Am J Trop Med Hyg 2007; 76:743-751.
9. Clements DE, Coller BA, Lieberman MM, Ogata S, Wang G, Harada KE, et al. Development of a recombinant tetravalent dengue virus vaccine: immunogenicity and efficacy studies in mice and monkeys. Vaccine 2010; 28:2705-2715.
10. Honda ER, Zanchi F, Rios K, Lira E, DeusileneVieira, da Silva LH, et al. Design and heterologous expression of dengue virus envelope protein (E) peptides and their use for serological diagnosis. J Virol Methods 2012; 186:55-61.
11. Clements DE, Coller BA, Lieberman MM, Ogata S, Wang G, Harada KE, et al. Expression of dengue-3 premembrane and envelope polyprotein in lettuce chloroplasts. Plant Mol Biol 2011; 76:323-333.
12. Modis Y, Ogata S, Clements D, Harrison SC. A ligand-binding pocket in the dengue virus envelope glycoprotein. Proc Natl Acad Sci USA 2003; 100:6986-6991.
13. Crill WD, Roehrig JT. Monoclonal antibodies that bind to domain III of dengue virus E glycoprotein are the most efficient blockers of virus adsorption to Vero cells. J Virol 2001; 75:7769-7773.
14. Hung JJ, Hsieh MT, Young MJ, Kao CL, King CC  Chang W. An external loop region of domain III of dengue virus type 2 envelope protein is involved in serotype-specific binding to mosquito but not mammalian cells. J Virol 2004; 78:378-388.
15. Chin JF, Chu JJ, Ng ML. The envelope glycoprotein domain III of dengue virus serotypes 1 and 2 inhibit virus entry. Microbes Infect 2007; 9:1-6.
16. Chen Y, Maguire T, Hileman RE, Fromm JR, Esko JD, Linhardt RJ, et al. Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nat Med 1997; 3 866-781.
17. Mune M, Rodriguez R, Ramirez R, Soto Y, Sierra B, Rodriguez Roche R, et al. Carboxy-terminally truncated Dengue 4 virus envelope glycoprotein expressed in Pichia pastoris induced neutralizing antibodies and resistance to Dengue 4 virus challenge in mice. Arch Virol 2003; 148:2267-2273.
18. Apt D, Raviprakash K, Brinkman A, Semyonov A, Yang S, Skinner C, et al. Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen. Vaccine 2006; 24:335-344.
19. Brandler S, Lucas-Hourani M, Moris A, Frenkiel MP, Combredet C, Fevrier M, et al. Pediatric measles vaccine expressing a dengue antigen induces durable serotype-specific neutralizing antibodies to dengue virus. PLoS Negl Trop Dis 2007; 1:e96.
20. Leng CH, Liu SJ, Tsai JP, Li YS, Chen MY, Liu HH, et al. A novel dengue vaccine candidate that induces cross-neutralizing antibodies and memory immunity. Microbes Infect 2009; 11:288-295.
21. Khanam S, Khanna N, Swaminathan S. Induction of neutralizing antibodies and T cell responses by dengue virus type 2 envelope domain III encoded by plasmid and adenoviral vectors. Vaccine 2006; 24:6513-6525.
22. Khanam S, Rajendra P, Khanna N, Swaminathan S. An adenovirus prime/plasmid boost strategy for induction of equipotent immune responses to two dengue virus serotypes. BMC Biotechnol 2007; 7:10.
23. Etemad B, Batra G, Raut R, Dahiya S, Khanam S, Swaminathan S, et al. An envelope domain III-based chimeric antigen produced in Pichia pastoris elicits neutralizing antibodies against all four dengue virus serotypes. Am J Trop Med Hyg 2008; 79:353-363.
24. Jaiswal S, Khanna N, Swaminathan S. High-level expression and one-step purification of recombinant dengue virus type 2 envelope domain III protein in Escherichia coli. Protein Expr Purif 2004; 33:80-91.
25. Liao M, Kielian M. Domain III from class II fusion proteins functions as a dominant-negative inhibitor of virus membrane fusion. J Cell Biol 2005; 171:111-120.
26. Babu JP, Pattnaik P, Gupta N, Shrivastava A, Khan M, Rao PV. Immunogenicity of a recombinant envelope domain III protein of dengue virus type-4 with various adjuvants in mice. Vaccine 2008; 26: 4655-4663.
27. Prinz WA, Aslund F, Holmgren A, Beckwith J. The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm. J Biol Chem 1997; 272:15661-1567.
28. Bessette PH, Aslund F, Beckwith J, Georgiou G. Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm. PNAS 1999; 96: 13703–13708.
 
29. Swaminathan S, Khanna N. Dengue vaccine – current progress and challenges. Current Science, Special Section: Biology and Pathogenesis of Viruses 2010; 98:3,369-378.
30. Goeddel DV. Systems for heterologous gene expression. Methods Enzymol 1990; 185:3-7.
31. Burgess-Brown NA, Sharma S, Sobott F, Loenarz C, Oppermann U, Gileadi O. Codon optimization can improve expression of human genes in Escherichia coli: A multi-gene study. Protein Expr Purif 2008; 59:94-102.
32. Simmons M, Nelson WM, Wu SJ, Hayes CG. Evaluation of the protective efficacy of a recombinant dengue envelope B domain fusion protein against dengue 2 virus infection in mice. Am J Trop Med Hyg 1998; 58:655-662.
 
Iranian Journal of Basic Medical Sciences
Volume 17, Issue 11 - Serial Number 11
November 2014
Pages 836-843

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