Volume 69, Issue 2
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


A recombinant vaccine that expresses the envelope (E) gene of dengue virus type 4 was tested for immunogenicity and protection in . One hundred micrograms of semipurified recombinant E protein (E4rec) expressed in was used to immunize three animals. Neutralizing antibodies to dengue 4 virus with a titer of 1:30 were detected in all immunized monkeys prior to challenge. Animals were challenged with 10 plaque-forming units of dengue 4 virus. One vaccine-immunized monkey was protected from viremia, while the other two were partially protected. Monkeys immunized with E4rec elicited the highest neutralizing antibody titers ( < 0.05) ranging from 1:85 to 1:640 at day 30. In both immunized and control animals, the longest duration of viremia correlated with earliest and highest level of IgM antibody to dengue virus. The vaccinated animals showed anamnestic antibody responses upon virus challenge, indicating successful priming by the recombinant vaccine. Our results suggest that E4rec expressed in can provide partial protection against viremia. However, the results were not effective enough to use it as a vaccine candidate. Further work is required to improve the quality of the immunogen.


Article metrics loading...

The graphs shown below represent data from March 2017
Loading full text...

Full text loading...



  1. Gubler DJ, 2002. Epidemic dengue/dengue hemorrhagic fever as a public health social and economic problem in the 21st century. Trends Microbiol 10 : 100–103. [Google Scholar]
  2. Guzman MG, Kouri G, 2002. Dengue, an update. Lancet Infect Dis 2 : 33–42. [Google Scholar]
  3. Chambers TJ, Hahn C, Galler R, Rice CM, 1990. Flavivirus genome organization, expression and replication. Annu Rev Microbiol 44 : 649–688. [Google Scholar]
  4. Putnak R, 1994. Progress in the development of recombinant vaccines against dengue and other arthropod-borne flaviviruses. Kurstak E, ed. Modern Vaccinology. New York: Plenum Medical, 231–252.
  5. Roehrig JT, 1997. Immunochemistry of dengue viruses. Gubler D, Kuno G, eds. Dengue and Dengue Hemorrhagic Fever. New York: CAB International, 199–219.
  6. Chambers TJ, Tsai TF, Pervikov Y, Monath TP, 1997. Vaccine development against dengue and Japanese encephalitis: report of a World Health Organization meeting. Vaccine 15 : 1494–1502. [Google Scholar]
  7. Bhamarapravati N, Sutee Y, 2000. Live attenuated tetravalent dengue vaccine. Vaccine 18 : 44–47. [Google Scholar]
  8. Men R, Wyatt L, Tokimatsu I, Arakaki S, Shameem G, Elkins R, Chanock R, Moss B, Lai CJ, 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. [Google Scholar]
  9. Huang C, Butrapet S, Pierro DJ, Chang GJ, Hunt AR, Bhamarapravati N, Gubler D, Kinney RM, 2000. Chimeric dengue type 2 (vaccine strain PDK-53)/dengue type 1 as potential candidate dengue type 1 virus vaccine. J Virol 74 : 3020–3028. [Google Scholar]
  10. Kochel TJ, Raviprakash K, Hayes CG, Watts DM, Rusell KL, Gozalo AS, Phillips IA, Ewing DF, Murphy GS, Porter KR, 2000. A dengue virus serotype-1 DNA vaccine induces virus neutralizing antibodies and provides protection from viral challenge in Aotus monkeys. Vaccine 18 : 3166–3173. [Google Scholar]
  11. Putnak R, Barvir DA, Burrous JM, Dubois DR, Dándrea VM, Hoke CH, Sadoff JC, Eckels KH, 1996. Development of a purified, inactivated, dengue-2 virus vaccine prototype in Vero cells: immunogenicity and protection in mice and rhesus monkeys. J Infect Dis 174 : 1176–1184. [Google Scholar]
  12. Bielefeldt-Ohmann H, Beasley DWC, Fitspatrick DR, Aaskov JG, 1997. Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretory baculovirus system. J Gen Virol 78 : 2723–2733. [Google Scholar]
  13. Sugrue RJ, Fu J, Howe J, Chan Y-C, 1997. Expression of the dengue virus structural proteins in Pichia pastoris leads to the generation of virus-like particles. J Gen Virol 78 : 1861–1866. [Google Scholar]
  14. Sugrue RJ, Cui T, Xu Q, Fu J, Chan YC, 1997. The production of recombinant dengue virus E protein using Escherichia coli and Pichia pastoris. J Virol Methods 69 : 159–169. [Google Scholar]
  15. Bisht H, Chugh DA, Raje M, Swaminathan, Khanna N, 2002. Recombinant dengue virus type 2 envelope/hepatitis B surface antigen hybrid protein expressed in Pichia pastoris can function as a bivalent immunogen. J Biotechol 99 : 97–110. [Google Scholar]
  16. Guirakhoo F, Bolin RA, Roerhig JT, 1992. The Murray Valley Encephalitis virus prM protein confers acid resistance to virus particles and alters the expression of epitopes within the R2 domain of the E glygoprotein. Virology 191 : 921–931. [Google Scholar]
  17. Tschopp JF, Brust PE, Cregg JM, Stillman CA, Gingeras GR, 1987. Expression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris. Nucleic Acids Res 15 : 3859–3876. [Google Scholar]
  18. Men R, Bray M, Lai CJ, 1991. Carboxy-terminally truncated dengue virus envelope glycoproteins expressed on the cell surface and secreted extracellularly exhibit increased immunogenicity in mice. J Virol 65 : 1400–1407. [Google Scholar]
  19. Hermida L, Rodriguez R, Lazo L, Lopez C, Marquez G, Paez R, Guzman MG, Suarez C, Espinosa R, Garcia J, Guillen G, 2002. A recombinant envelope protein from dengue virus purified by IMAC is bioequivalent with its immune-affinity chromatography purified counterpart. J Biotechnol 94 : 213–216. [Google Scholar]
  20. Morens DM, Halstead SB, Repik PM, Putvatana R, Raybourne N, 1985. Simplified plaque reduction neutralization assay for dengue viruses by semimicro methods in BHK21-cells: comparison of the BHK suspension test with the standard plaque reduction neuralization. J Clin Microbiol 22 : 250–254. [Google Scholar]
  21. Vazquez S, Saenz E, Huelva G, González A, Kouri G, Guzman MG, 1998. Detección de IgM contra el virus dengue en sangre total absorbida en papel de filtro. Rev Panam Salud Publica 3 : 174–178. [Google Scholar]
  22. Pupo M, Rodriguez R, Alvarez M, Amin N, Rodriguez H, Otero A, Guzman MG, 2001. Development of a monoclonal antibody specific to a recombinant envelope protein from dengue virus type 4 expressed in Pichia pastoris. Hybridoma 20 : 35–41. [Google Scholar]
  23. Clarke DH, Casals J, 1958. Techniques for hemagglutination and hemmaglutination-inhibition with arthropod-borne viruses. Am J Trop Med Hyg 7 : 561–573. [Google Scholar]
  24. Rodríguez R, Alvarez M, Guzman MG, Morier L, Kouri G, 2000. Isolation of dengue 2 virus in C636/HT cells by rapid centrifugation/shell vial assay: comparison with conventional virus isolation method. J Clin Microbiol 38 : 3508–3510. [Google Scholar]
  25. Lanciotti RS, Calisher CH, Gubler DJ, Chang GJ, Vorndam AV, 1992. Rapid detection and typing of dengue viruses from clinical samples using reverse transcriptase-polymerase chain reaction. J Clin Microbiol 30 : 545–551. [Google Scholar]
  26. Rosario D, Alvarez M, Díaz J, Contreras R, Vázquez S, Rodriguez R, Guzmán MG, 1998. Rapid detection and typing of dengue viruses from clinical samples using reverse transcriptase-polymerase chain reaction (in Spanish). Rev Panam Salud Publica 4 : 1–5. [Google Scholar]
  27. Feighny R, Burrous J, Putnak R, 1994. Dengue type 2 virus envelope protein made using recombinant baculovirus protects mice against virus challenge. Am J Trop Med Hyg 50 : 322–328. [Google Scholar]
  28. Scott RM, Nisalak A, Eckels KH, Tingpalapong M, Harrison VR, Gould DJ, Chapple FE, Russell PK, 1980. Dengue-2 vaccine: viremia and immune responses in rhesus monkeys. Infect Immun 27 : 181–186. [Google Scholar]
  29. Raviprakash K, Porter KR, Kochel TJ, Ewing D, Simmons M, Phillips I, Murphy GS, Weiss WR, Hayes CG, 2000. Dengue virus type 1 DNA vaccine induces protective immune responses in rhesus macaques. J Gen Virol 81 : 1659–1667. [Google Scholar]

Data & Media loading...

  • Received : 30 Sep 2002
  • Accepted : 12 Apr 2003

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error