Volume 76, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


There are approximately 100 million new cases of dengue (DEN) virus infection each year. Infection can result in illness ranging from a mild fever to hemorrhaging, shock, or even death. There are four serotypes of dengue virus (DEN1–4), and immunity to one serotype does not cross protect from infection with other serotypes. Currently there are no approved vaccines for dengue fever. In this report, we describe the construction of a bivalent dengue virus vaccine using a complex recombinant adenovirus approach to express multiple genes of DEN1 and DEN2 serotypes. In vaccinated mice, this vector induced humoral immune responses against all four dengue serotypes as measured by enzyme-linked immunosorbent assay. However, the neutralizing antibody responses were specific for DEN1 and DEN2 serotypes. Expansion of this vaccine development platform towards the DEN3 and DEN4 serotypes can lead towards the development of an adenovirus-based tetravalent dengue vaccine.


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  1. Stephenson JR, 2005. Understanding dengue pathogenesis: implications for vaccine design. Bull World Health Organ 83 : 308–314. [Google Scholar]
  2. Gubler DJ, 1998. Dengue and dengue hemorrhagic fever. Clin Microbiol Rev 11 : 480–496. [Google Scholar]
  3. Gubler DJ, 1998. Resurgent vector-borne diseases as a global health problem. Emerg Infect Dis 4 : 442–450. [Google Scholar]
  4. Henchal EA, Putnak JR, 1990. The dengue viruses. Clin Microbiol Rev 3 : 376–396. [Google Scholar]
  5. Edelman R, Wasserman SS, Bodison SA, Putnak RJ, Eckels KH, Tang D, Kanesa-Thasan N, Vaughn DW, Innis BL, Sun W, 2003. Phase I trial of 16 formulations of a tetravalent live-attenuated dengue vaccine. Am J Trop Med Hyg 69 : 48–60. [Google Scholar]
  6. Kanesa-Thasan N, Edelman R, Tacket CO, Wasserman SS, Vaughn DW, Coster TS, Kim-Ahn GJ, Dubois DR, Putnak JR, King A, Summers PL, Innis BL, Eckels KH, Hoke CH Jr, 2003. Phase 1 studies of Walter Reed Army Institute of Research candidate attenuated dengue vaccines: selection of safe and immunogenic monovalent vaccines. Am J Trop Med Hyg 69 : 17–23. [Google Scholar]
  7. Kinney RM, Butrapet S, Chang GJ, Tsuchiya KR, Roehrig JT, Bhamarapravati N, Gubler DJ, 1997. Construction of infectious cDNA clones for dengue 2 virus: strain 16681 and its attenuated vaccine derivative, strain PDK-53. Virology 230 : 300–308. [Google Scholar]
  8. Sun W, Edelman R, Kanesa-Thasan N, Eckels KH, Putnak JR, King AD, Houng HS, Tang D, Scherer JM, Hoke CH Jr, Innis BL, 2003. Vaccination of human volunteers with monovalent and tetravalent live-attenuated dengue vaccine candidates. Am J Trop Med Hyg 69 : 24–31. [Google Scholar]
  9. Bielefeldt-Ohmann H, Beasley DW, Fitzpatrick 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]
  10. Kelly EP, Greene JJ, King AD, Innis BL, 2000. Purified dengue 2 virus envelope glycoprotein aggregates produced by baculovirus are immunogenic in mice. Vaccine 18 : 2549–2559. [Google Scholar]
  11. Putnak R, Coller BA, Voss G, Vaughn DW, Clements D, Peters I, Bignami G, Houng HS, Chen RC, Barvir DA, Seriwatana J, Cayphas S, Garcon N, Gheysen D, Kanesa-Thasan N, McDonell M, Humphreys T, Eckels KH, Prieels JP, Innis BL, 2005. An evaluation of dengue type-2 inactivated, recombinant subunit, and live-attenuated vaccine candidates in the rhesus macaque model. Vaccine 23 : 4442–4452. [Google Scholar]
  12. Velzing J, Groen J, Drouet MT, van Amerongen G, Copra C, Osterhaus AD, Deubel V, 1999. Induction of protective immunity against Dengue virus type 2: comparison of candidate live attenuated and recombinant vaccines. Vaccine 17 : 1312–1320. [Google Scholar]
  13. Bray M, Lai CJ, 1991. Construction of intertypic chimeric dengue viruses by substitution of structural protein genes. Proc Natl Acad Sci U S A 88 : 10342–10346. [Google Scholar]
  14. Bray M, Men R, Lai CJ, 1996. Monkeys immunized with intertypic chimeric dengue viruses are protected against wild-type virus challenge. J Virol 70 : 4162–4166. [Google Scholar]
  15. Huang CY, Butrapet S, Tsuchiya KR, Bhamarapravati N, Gubler DJ, Kinney RM, 2003. Dengue 2 PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development. J Virol 77 : 11436–11447. [Google Scholar]
  16. Chambers TJ, Liang Y, Droll DA, Schlesinger JJ, Davidson AD, Wright PJ, Jiang X, 2003. Yellow fever virus/dengue-2 virus and yellow fever virus/dengue-4 virus chimeras: biological characterization, immunogenicity, and protection against dengue encephalitis in the mouse model. J Virol 77 : 3655–3668. [Google Scholar]
  17. Guirakhoo F, Weltzin R, Chambers TJ, Zhang ZX, Soike K, Ratterree M, Arroyo J, Georgakopoulos K, Catalan J, Monath TP, 2000. Recombinant chimeric yellow fever-dengue type 2 virus is immunogenic and protective in nonhuman primates. J Virol 74 : 5477–5485. [Google Scholar]
  18. Kochel T, Wu SJ, Raviprakash K, Hobart P, Hoffman S, Porter K, Hayes C, 1997. Inoculation of plasmids expressing the dengue-2 envelope gene elicit neutralizing antibodies in mice. Vaccine 15 : 547–552. [Google Scholar]
  19. Konishi E, Yamaoka M, Kurane I, Mason PW, 2000. A DNA vaccine expressing dengue type 2 virus premembrane and envelope genes induces neutralizing antibody and memory B cells in mice. Vaccine 18 : 1133–1139. [Google Scholar]
  20. 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]
  21. Mota J, Acosta M, Argotte R, Figueroa R, Mendez A, Ramos C, 2005. Induction of protective antibodies against dengue virus by tetravalent DNA immunization of mice with domain III of the envelope protein. Vaccine 23 : 3469–3476. [Google Scholar]
  22. Apt D, Raviprakash K, Brinkman A, Semyonov A, Yang S, Skinner C, Diehl L, Lyons R, Porter K, Punnonen J, 2006. Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen. Vaccine 24 : 335–344. [Google Scholar]
  23. Kochel TJ, Raviprakash K, Hayes CG, Watts DM, Russell 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]
  24. Putnak R, Fuller J, VanderZanden L, Innis BL, Vaughn DW, 2003. Vaccination of rhesus macaques against dengue-2 virus with a plasmid DNA vaccine encoding the viral pre-membrane and envelope genes. Am J Trop Med Hyg 68 : 469–476. [Google Scholar]
  25. Arribillaga L, de Cerio AL, Sarobe P, Casares N, Gorraiz M, Vales A, Bruna-Romero O, Borras-Cuesta F, Paranhos-Baccala G, Prieto J, Ruiz J, Lasarte JJ, 2002. Vaccination with an adenoviral vector encoding hepatitis C virus (HCV) NS3 protein protects against infection with HCV-recombinant vaccinia virus. Vaccine 21 : 202–210. [Google Scholar]
  26. Jacobs SC, Stephenson JR, Wilkinson GW, 1992. High-level expression of the tick-borne encephalitis virus NS1 protein by using an adenovirus-based vector: protection elicited in a murine model. J Virol 66 : 2086–2095. [Google Scholar]
  27. Jaiswal S, Khanna N, Swaminathan S, 2003. Replication-defective adenoviral vaccine vector for the induction of immune responses to dengue virus type 2. J Virol 77 : 12907–12913. [Google Scholar]
  28. Cohen P, 2006. Immunity’s yin and yang. A successful vaccine must first avoid being eliminated by pre-existing immunity before it can promote a protective immune response. IAVI Rep 10 : 1–5. [Google Scholar]
  29. van Kampen KR, Shi Z, Gao P, Zhang J, Foster KW, Chen DT, Marks D, Elmets CA, Tang DC, 2005. Safety and immunogenicity of adenovirus-vectored nasal and epicutaneous influenza vaccines in humans. Vaccine 23 : 1029–1036. [Google Scholar]
  30. Wang D, Hevey M, Juompan LY, Trubey CM, Raja NU, Deitz SB, Woraratanadharm J, Luo M, Yu H, Swain BM, Moore KM, Dong JY, 2006. Complex adenovirus-vectored vaccine protects guinea pigs from three strains of Marburg virus challenges. Virology 353 : 324–332. [Google Scholar]
  31. Wang D, Schmaljohn AL, Raja NU, Trubey CM, Juompan LY, Luo M, Deitz SB, Yu H, Woraratanadharm J, Holman DH, Moore KM, Swain BM, Hart MK, Pratt WD, Dong JY, 2006. De novo syntheses of Marburg virus antigens from adenovirus vectors induce potent humoral and cellular immune responses. Vaccine 24 : 2975–2985. [Google Scholar]
  32. Wang D, Raja NU, Trubey CM, Juompan LY, Luo M, Woraratanadharm J, Deitz SB, Yu H, Swain BM, Moore KM, Pratt WD, Hart MK, Dong JY, 2006. Development of a cAdVax-based bivalent Ebola vaccine that induces immune responses against both the Sudan and Zaire species of Ebola virus. J Virol 80 : 2738–2746. [Google Scholar]
  33. Rubinchik S, Woraratanadharm J, Schepp J, Dong JY, 2003. Improving the transcriptional regulation of genes delivered by adenovirus vectors. Methods Mol Med 76 : 167–199. [Google Scholar]
  34. Rubinchik S, Norris JS, Dong JY, 2002. Construction, purification and characterization of adenovirus vectors expressing apoptosis-inducing transgenes. Methods Enzymol 346 : 529–547. [Google Scholar]
  35. Henchal EA, Gentry MK, McCown JM, Brandt WE, 1982. Dengue virus-specific and flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence. Am J Trop Med Hyg 31 : 830–836. [Google Scholar]
  36. Henchal EA, McCown JM, Seguin MC, Gentry MK, Brandt WE, 1983. Rapid identification of dengue virus isolates by using monoclonal antibodies in an indirect immunofluorescence assay. Am J Trop Med Hyg 32 : 164–169. [Google Scholar]
  37. Crill WD, Roehrig JT, 2001. 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 75 : 7769–7773. [Google Scholar]
  38. Mathew A, Kurane I, Green S, Stephens HA, Vaughn DW, Kalayanarooj S, Suntayakorn S, Chandanayingyong D, Ennis FA, Rothman AL, 1998. Predominance of HLA-restricted cytotoxic T-lymphocyte responses to serotype-cross-reactive epitopes on nonstructural proteins following natural secondary dengue virus infection. J Virol 72 : 3999–4004. [Google Scholar]
  39. Hanna SL, Pierson TC, Sanchez MD, Ahmed AA, Murtadha MM, Doms RW, 2005. N-linked glycosylation of west nile virus envelope proteins influences particle assembly and infectivity. J Virol 79 : 13262–13274. [Google Scholar]
  40. Konishi E, Mason PW, 1993. Proper maturation of the Japanese encephalitis virus envelope glycoprotein requires cosynthesis with the premembrane protein. J Virol 67 : 1672–1675. [Google Scholar]
  41. Simmons M, Murphy GS, Hayes CG, 2001. Short report: Antibody responses of mice immunized with a tetravalent dengue recombinant protein subunit vaccine. Am J Trop Med Hyg 65 : 159–161. [Google Scholar]
  42. Jan LR, Yang CS, Henchal LS, Sumiyoshi H, Summers PL, Dubois DR, Lai CJ, 1993. Increased immunogenicity and protective efficacy in outbred and inbred mice by strategic carboxyl-terminal truncation of Japanese encephalitis virus envelope glycoprotein. Am J Trop Med Hyg 48 : 412–423. [Google Scholar]
  43. Livingston PG, Kurane I, Lai CJ, Bray M, Ennis FA, 1994. Recognition of envelope protein by dengue virus serotype-specific human CD4+ CD8- cytotoxic T-cell clones. J Virol 68 : 3283–3288. [Google Scholar]
  44. Yang ZY, Duckers HJ, Sullivan NJ, Sanchez A, Nabel EG, Nabel GJ, 2000. Identification of the Ebola virus glycoprotein as the main viral determinant of vascular cell cytotoxicity and injury. Nat Med 6 : 886–889. [Google Scholar]
  45. Kitchener S, Nissen M, Nasveld P, Forrat R, Yoksan S, Lang J, Saluzzo JF, 2006. Immunogenicity and safety of two live-attenuated tetravalent dengue vaccine formulations in healthy Australian adults. Vaccine 24 : 1238–1241. [Google Scholar]

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  • Received : 26 Apr 2006
  • Accepted : 18 Nov 2006

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