Health  Vol.2 No.11 , November 2010
Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus
ABSTRACT
In this work, we report the evaluation of two DNA vaccines against dengue-3 virus (DENV-3). The first construction, called pVAC3DEN3, was engineered inserting the pre-membrane (prM) and envelope (E) gene of DENV-3 truncated with a restriction site between them, as previously described. The second construction was developed cloning the full gene sequence of prM and E from DENV-3 virus in pCI plasmid for mammalian expression and was denominated pVAC1WDEN3. The results showed that both constructions were capable of expressing the prM and E proteins, as demonstrated by ELISA and immunoblotting detection in cell culture transfected with the plasmids. After positive “in vitro” results, the vaccine candidates were used to immunize BALB/c mice and the elicited response was investigated. After immunization by intramuscular inoculation with three doses of each vaccinal clone the animals were sacrificed, the cytokine levels and T cell response were analyzed in the spleens, after three days of culture with stimulus, our analysis showed that the two constructions elicited T cell responses mea- sured by BrdU incorporation assay and high levels of IFN-γ, detected in the supernatant of the cultures. Moreover, both constructions induced detectable titers of neutralizing antibodies in mice. And finally the survival rate of the immunized animals after intracerebral challenge was analyzed, showing a better result in the pVAC3DEN3 group with an 80% survival compared with a 50% survival of the pVAC1 WDEN3. Thus, these data showed that our two constructions were able to induce specific immune response and protects mice against a lethal challenge with DENV-3, and these vaccine candidates can be employed to develop a viable dengue vaccine.

Cite this paper
nullPaula, S. , França, R. , Lima, D. , Dutra, N. , Paula, M. , Oliveira, M. , Oliveira, L. and Fonseca, B. (2010) Evaluation of immunogenicity elicited from two DNA vaccine candidates that expresses the prM and E genes of the dengue-3 virus. Health, 2, 1298-1307. doi: 10.4236/health.2010.211193.
References
[1]   Whitehead, S.S., Blaney, J.E., Durbin, A.P. and Murphy, B.R. (2007) Prospects for a dengue virus vaccine. Nature Reviews Microbiology, 5, 518-528.

[2]   Halstead, S.B. (1989) Antibody, macrophages, dengue virus-infection, shock, and hemorrhage: A pathogenetic cascade. Reviews of Infectious Diseases, 11, S830-S839.

[3]   Innis, B.L. (1995) Dengue and dengue hemorrhagic fever. In: Porterfield, J.S. Ed., Exotic Viral Infections, Chapman Hall, London, 103-146.

[4]   Kuno, G. (2004) Serodiagnosis of flaviviral infections and vaccinations in humans. Advances in Virus Research, 61, 63-65.

[5]   Gubler, D.J. (1998) Population growth, urbanization, au- tomobiles and aeroplanes: The dengue connection. In: Greenwood, B. and De Cock, K. Eds., New & Resurgent Infection. Prediction, Detection & Management of Tomorrows Epidemics, John Wiley & Sons, New York, 117- 129.

[6]   Gubler, D.J. (2002) The global emergence/resurgence of arboviral diseases as public health problems. Archives of Medical Research, 33, 330-342.

[7]   Barrett, A.D.T. (1997) Yellow fever vaccines. Biologicals, 25, 17-25.

[8]   Barrett, A.D.T. (1997) Japanese encephalitis and dengue vaccines. Biologicals, 25, 27-34.

[9]   Raviprakash, K., Kochel, T.J., Ewing, D., Simmons, M., Phillips, I., et al. (2000) Immunogenicity of dengue virus type 1 DNA vaccines expressing truncated and full length envelope protein. Vaccine, 18, 2426-2434.

[10]   Edelman, R. (2005) Dengue and dengue vaccines. Journal of Infectious Diseases, 191, 650-653.

[11]   Kinney, R.M. and Huang, C.Y.H. (2001) Development of new vaccines against dengue fever and Japanese encephalitis. Intervirology, 44, 176-197.

[12]   Imoto, J., Konishi, E. (2007) Dengue tetravalent DNA vaccine increases its immunogenicity in mice when mixed with a dengue type 2 subunit vaccine or an inactivated Japanese encephalitis vaccine. Vaccine, 25, 1076- 1084.

[13]   Colombage, G., Hall, R., Pavy, M. and Lobigs, M. (1998) DNA-based and alphavirus-vectored immunisation with prM and E proteins elicits long-lived and protective immunity against the flavivirus, Murray Valley encephalitis virus. Virology, 250, 151-163.

[14]   Konishi, E., Yamaoka, M., Khin Sane, W., Kurane, I. and Mason, P.W. (1998) Induction of protective immunity against Japanese encephalitis in mice by immunization with a plasmid encoding Japanese encephalitis virus premembrane and envelope genes. Journal of Virology, 72, 4925-4930.

[15]   Phillpotts, R.J., Venugopal, K. and Brooks, T. (1996) Immunisation with DNA polynucleotides protects mice against lethal challenge with St Louis encephalitis virus. Archives of Virology, 141, 743-749.

[16]   Schmaljohn, C., Vanderzanden, L., Bray, M., Custer, D., Meyer, B., et al. (1997) Naked DNA vaccines expressing the prM and E genes of Russian spring summer encephalitis virus and Central European encephalitis virus protect mice from homologous and heterologous challenge. Jour- nal of Virology, 71, 9563-9569.

[17]   Alves, A.M.B., Lasaro, M.O., Almeida, D.F. and Ferreira, L.C.S. (1999) New vaccine strategies against enterotoxigenic Escherichia coli. I: DNA vaccines against the CFA/I fimbrial adhesin. Brazilian Journal of Medical and Biological Research, 32, 223-229.

[18]   Donnelly, J.J., Wahren, B. and Liu, M.A. (2005) DNA vaccines: Progress and challenges. Journal of Immunology, 175, 633-639.

[19]   Mukhopadhyay, S., Kuhn, R.J., Rossmann, M.G. (2005) A structural perspective of the Flavivirus life cycle. Nature Reviews Microbiology, 3, 13-22.

[20]   Kuhn, R.J., Zhang, W., Rossmann, M.G., Pletnev, S.V., Corver, J., et al. (2002) Structure of dengue virus: Implications for flavivirus organization, maturation, and fusion. Cell, 108, 717-725.

[21]   Brinton, M.A., Kurane, I., Mathew, A., Zeng, L.L., Shi, P.Y., et al. (1998) Immune mediated and inherited defences against flaviviruses. Clinical and Diagnostic Virology, 10, 129-139.

[22]   Chambers, T.J., Hahn, C.S., Galler, R. and Rice, C.M. (1990) Flavivirus genome organization, expression, and replication. Annual Review of Microbiology, 44, 649-688.

[23]   De Paula, S.O., Lima, D.M., Oliveira Fran?a, R.F., Gomes-Ruiz, A.C. and Fonseca, B.A.L (2008) A DNA vaccine candidate expressing dengue-3 virus prM and E pro- teins elicits neutralizing antibodies and protects mice against lethal challenge. Archives of Virology. 153(12), 2215-2223.

[24]   Russell, P.K. and Nisalak, A. (1967) Dengue virus identification by plaque reduction neutralization test. Journal of Immunology, 99, 291.

[25]   Konishi, E., Pincus, S., Paoletti, E., Shope, R.E., Burrage, T., et al. (1992) Mice immunized with a subviral particle containing the Japanese Encephalitis-virus prM/M and E-proteins are protected from lethal JEV infection. Virology, 188, 714-720.

[26]   Fonseca, B.A.L., Pincus, S., Shope, R.E., Paoletti, E. and Mason, P.W. (1994) Recombinant Vaccinia viruses co- expressing Dengue-1 glycoproteins prM and E-induce neutralizing antibodies in mice. Vaccine, 12, 279-285.

[27]   Allison, S.L., Stadler, K., Mandl, C.W., Kunz, C., Heinz, F.X. (1995) Synthesis and secretion of recombinant Tick- Borne Encephalitis-Virus protein-E in soluble and particulate form. Journal of Virology, 69, 5816-5820.

[28]   Lorenz, I.C., Allison, S.L., Heinz, F.X., Helenius, A. (2002) Folding and dimerization of tick-borne encephalitis virus envelope proteins prM and E in the endoplasmic reticulum. Journal of Virology, 76, 5480-5491.

[29]   Jimenez, R.O. and da Fonseca, B.A.L. (2000) Recombinant plasmid expressing a truncated dengue-2 virus E protein without co-expression of prM protein induces partial protection in mice. Vaccine, 19, 648-654.

[30]   Men, R., Wyatt, L., Tokimatsu, I., Arakaki, S., Shameem, G., et al. (2000) Immunization of rhesus monkeys with a recombinant of modified vaccinia virus Ankara expressing a truncated envelope glycoprotein of dengue type 2 virus induced resistance to dengue type 2 virus challenge. Vaccine, 18, 3113-3122.

[31]   Guirakhoo, F., Pugachev, K., Zhang, Z., Myers, G. and Levenbook, I., et al. (2004) Safety and efficacy of chimeric yellow fever-dengue virus tetravalent vaccine formulations in nonhuman primates. Journal of Virology, 78, 4761-4775.

[32]   Khanam, S., Khanna, N. and Swarninathan, S. (2006) Induction of neutralizing antibodies and T cell responses by dengue virus type 2 envelope domain III encoded by plasmid and adenoviral vectors. Vaccine, 24, 6513-6525.

[33]   Bente, D.A. and Rico-Hesse, R. (2006) Models of dengue virus infection. Drug Discovery Today: Disease Mo- dels, 3(1), 97-103.

[34]   Kaufman, B.M., Summers, P.L., Dubois, D.R. and Eckels, K.H. (1987) Monoclonal-antibodies against Dengue-2 virus E-glycoprotein protect mice against lethal dengue infection. American Journal of Tropical Medicine and Hygiene, 36, 427-434.

[35]   Bray, M., Zhao, B.T., Markoff, L., Eckels, K.H., Cha- nock, R.M., et al. (1989) Mice immunized with recombinant vaccinia virus expressing Dengue-4 virus structural proteins with or without nonstructural protein-NS1 are protected against fatal Dengue virus encephalitis. Journal of Virology, 63, 2853-2856.

[36]   Falgout, B., Bray, M., Schlesinger, J.J. and Lai, C.J. (1990) Immunization of mice with recombinant Vaccinia virus expressing authentic Dengue virus nonstructural protein NS1 protects against lethal Dengue virus encephalitis. Journal of Virology, 64, 4356-4363.

[37]   Van Der Most, R.G. and Strauss, J.H. (2000) Chimeric yellow fever/dengue virus as a candidate dengue vaccine: Quantitation of the dengue virus-specific CD8 T-cell response. Journal of Virology, 74, 8094-8101.

[38]   Yauch, L.E. and Shresta, S. (2008) Mouse models of den- gue virus infection and disease. Antiviral Research, 80, 87-93.

 
 
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