Volume 92, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



The immunogenicity and protective efficacy of a candidate tetravalent dengue virus purified inactivated vaccine (TDENV PIV) formulated with alum or an Adjuvant System (AS01, AS03 tested at three different dose levels, or AS04) was evaluated in a 0, 1-month vaccination schedule in rhesus macaques. One month after dose 2, all adjuvanted formulations elicited robust and persisting neutralizing antibody titers against all four dengue virus serotypes. Most of the formulations tested prevented viremia after challenge, with the dengue serotype 1 and 2 virus strains administered at 40 and 32 weeks post-dose 2, respectively. This study shows that inactivated dengue vaccines, when formulated with alum or an Adjuvant System, are candidates for further development.

[open-access] This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Article metrics loading...

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

Full text loading...



  1. Monath TP, Fields BN, Knipe DM, Chanock RM, , 1990. Flaviviruses. , eds. Field's Virology, 2nd Ed. New York, NY: Raven Press, 763814. [Google Scholar]
  2. Ye J, Zhu B, Fu ZF, Chen H, Cao S, , 2013. Immune evasion strategies of flaviviruses. Vaccine 31: 461471.[Crossref] [Google Scholar]
  3. Perera R, Kuhn RJ, , 2008. Structural proteomics of dengue virus. Curr Opin Microbiol 11: 369377.[Crossref] [Google Scholar]
  4. Mukhopadhyay S, Kuhn RJ, Rossmann MG, , 2005. A structural perspective of the flavivirus life cycle. Nat Rev Microbiol 3: 1322.[Crossref] [Google Scholar]
  5. Bhatt S, Gething PW, Brady OJ, Messina JP, Farlow AW, Moyes CL, Drake JM, Brownstein JS, Hoen AG, Sankoh O, Myers MF, George DB, Jaenisch T, Wint GR, Simmons CP, Scott TW, Farrar JJ, Hay SI, , 2013. The global distribution and burden of dengue. Nature 496: 504507.[Crossref] [Google Scholar]
  6. Gubler DJ, , 1998. Dengue and dengue hemorrhagic fever. Clin Microbiol Rev 11: 480496. [Google Scholar]
  7. Hombach J, Cardosa MJ, Sabchareon A, Vaughn DW, Barrett AD, , 2007. Scientific consultation on immunological correlates of protection induced by dengue vaccines: report from a meeting held at the World Health Organization 17–18 November 2005. Vaccine 25: 41304139.[Crossref] [Google Scholar]
  8. Kou Z, Quinn M, Chen H, Rodrigo WW, Rose RC, Schlesinger JJ, Jin X, , 2008. Monocytes, but not T or B cells, are the principal target cells for dengue virus (DV) infection among human peripheral blood mononuclear cells. J Med Virol 80: 134146.[Crossref] [Google Scholar]
  9. Kurane I, Matsutani T, Suzuki R, Takasaki T, Kalayanarooj S, Green S, Rothman AL, Ennis FA, , 2011. T-cell responses to dengue virus in humans. Trop Med Health 39: 4551.[Crossref] [Google Scholar]
  10. Rothman AL, , 2010. Cellular immunology of sequential dengue virus infection and its role in disease pathogenesis. Curr Top Microbiol Immunol 338: 8398. [Google Scholar]
  11. Yoon IK, Rothman AL, Tannitisupawong D, Srikiatkhachorn A, Jarman RG, Aldstadt J, Nisalak A, Mammen MP, Jr Thammapalo S, Green S, Libraty DH, Gibbons RV, Getis A, Endy T, Jones JW, Koenraadt CJ, Morrison AC, Fansiri T, Pimgate C, Scott TW, , 2012. Underrecognized mildly symptomatic viremic dengue virus infections in rural Thai schools and villages. J Infect Dis 206: 389398.[Crossref] [Google Scholar]
  12. Alexander N, Balmaseda A, Coelho IC, Dimaano E, Hien TT, Hung NT, Jänisch T, Kroeger A, Lum LC, Martinez E, Siqueira JB, Thuy TT, Villalobos I, Villegas E, Wills B, , 2011. Multicentre prospective study on dengue classification in four South-east Asian and three Latin American countries. Trop Med Int Health 16: 936948.[Crossref] [Google Scholar]
  13. Yacoub S, Mongkolsapaya J, Screaton G, , 2013. The pathogenesis of dengue. Curr Opin Infect Dis 26: 284289.[Crossref] [Google Scholar]
  14. Lai CY, Tsai WY, Lin SR, Kao CL, Hu HP, King CC, Wu HC, Chang GJ, Wang WK, , 2008. Antibodies to envelope glycoprotein of dengue virus during the natural course of infection are predominantly cross-reactive and recognize epitopes containing highly conserved residues at the fusion loop of domain II. J Virol 82: 66316643.[Crossref] [Google Scholar]
  15. Dejnirattisai W, Jumnainsong A, Onsirisakul N, Fitton P, Vasanawathana S, Limpitikul W, Puttikhunt C, Edwards C, Duangchinda T, Supasa S, Chawansuntati K, Malasit P, Mongkolsapaya J, Screaton G, , 2010. Cross-reacting antibodies enhance dengue virus infection in humans. Science 328: 745748.[Crossref] [Google Scholar]
  16. Halstead SB, , 1970. Observations related to pathogensis of dengue hemorrhagic fever. VI. Hypotheses and discussion. Yale J Biol Med 42: 350362. [Google Scholar]
  17. Halstead SB, O'Rourke EJ, , 1977. Antibody-enhanced dengue virus infection in primate leukocytes. Nature 265: 739741.[Crossref] [Google Scholar]
  18. Durbin AP, Whitehead SS, , 2010. Dengue vaccine candidates in development. Curr Top Microbiol Immunol 338: 129143. [Google Scholar]
  19. Guy B, Guirakhoo F, Barban V, Higgs S, Monath TP, Lang J, , 2010. Preclinical and clinical development of YFV 17D-based chimeric vaccines against dengue, West Nile and Japanese encephalitis viruses. Vaccine 28: 632649.[Crossref] [Google Scholar]
  20. Sabchareon A, Wallace D, Sirivichayakul C, Limkittikul K, Chanthavanich P, Suvannadabba S, Jiwariyavej V, Dulyachai W, Pengsaa K, Wartel TA, Moureau A, Saville M, Bouckenooghe A, Viviani S, Tornieporth NG, Lang J, , 2012. Protective efficacy of the recombinant, live-attenuated, CYD tetravalent dengue vaccine in Thai schoolchildren: a randomised, controlled phase 2b trial. Lancet 380: 15591567.[Crossref] [Google Scholar]
  21. Falgout B, Bray M, Schlesinger JJ, Lai CJ, , 1990. Immunization of mice with recombinant vaccinia virus expressing authentic dengue virus nonstructural protein NS1 protects against lethal dengue virus encephalitis. J Virol 64: 43564363. [Google Scholar]
  22. Blaney JE, Jr Sathe NS, Goddard L, Hanson CT, Romero TA, Hanley KA, Murphy BR, Whitehead SS, , 2008. Dengue virus type 3 vaccine candidates generated by introduction of deletions in the 3′ untranslated region (3′-UTR) or by exchange of the DENV-3 3′-UTR with that of DENV-4. Vaccine 26: 817828.[Crossref] [Google Scholar]
  23. Kinney RM, Huang CY, , 2001. Development of new vaccines against dengue fever and Japanese encephalitis. Intervirology 44: 176197.[Crossref] [Google Scholar]
  24. Osorio JE, Brewoo JN, Silengo SJ, Arguello J, Moldovan IR, Tary-Lehmann M, Powell TD, Livengood JA, Kinney RM, Huang CY, Stinchcomb DT, , 2011. Efficacy of a tetravalent chimeric dengue vaccine (DENVax) in cynomolgus macaques. Am J Trop Med Hyg 84: 978987.[Crossref] [Google Scholar]
  25. Durbin AP, Whitehead SS, Shaffer D, Elwood D, Wanionek K, Thumar B, Blaney JE, Murphy BR, Schmidt AC, , 2011. A single dose of the DENV-1 candidate vaccine rDEN1Δ30 is strongly immunogenic and induces resistance to a second dose in a randomized trial. PLoS Negl Trop Dis 5: e1267.[Crossref] [Google Scholar]
  26. Durbin AP, Kirkpatrick BD, Pierce KK, Elwood D, Larsson CJ, Lindow JC, Tibery C, Sabundayo BP, Shaffer D, Talaat KR, Hynes NA, Wanionek K, Carmolli MP, Luke CJ, Murphy BR, Subbarao K, Whitehead SS, , 2013. A single dose of any of four different live attenuated tetravalent dengue vaccines is safe and immunogenic in flavivirus-naive adults: a randomized, double-blind clinical trial. J Infect Dis 207: 957965.[Crossref] [Google Scholar]
  27. Slifka MK, , 2014. Vaccine-mediated immunity against dengue and the potential for long-term protection against disease. Front Immunol 5: 195.[Crossref] [Google Scholar]
  28. Halstead SB, , 2013. Identifying protective dengue vaccines: guide to mastering an empirical process. Vaccine 31: 45014507.[Crossref] [Google Scholar]
  29. Sabchareon A, Lang J, Chanthavanich P, Yoksan S, Forrat R, Attanath P, Sirivichayakul C, Pengsaa K, Pojjaroen-Anant C, Chambonneau L, Saluzzo JF, Bhamarapravati N, , 2004. Safety and immunogenicity of a three dose regimen of two tetravalent live-attenuated dengue vaccines in five- to twelve-year-old Thai children. Pediatr Infect Dis J 23: 99109.[Crossref] [Google Scholar]
  30. Thomas SJ, Eckels KH, Carletti I, De la Barrera R, Dessy F, Fernandez S, Putnak R, Toussaint JF, Sun W, Bauer K, Gibbons RV, Innis BL, , 2013. A phase II, randomized, safety and immunogenicity study of a re-derived, live-attenuated dengue virus vaccine in healthy adults. Am J Trop Med Hyg 88: 7388.[Crossref] [Google Scholar]
  31. Tauber E, Kollaritsch H, von Sonnenburg F, Lademann M, Jilma B, Firbas C, Jelinek T, Beckett C, Knobloch J, McBride WJ, Schuller E, Kaltenböck A, Sun W, Lyons A, , 2008. Randomized, double-blind, placebo-controlled phase 3 trial of the safety and tolerability of IC51, an inactivated Japanese encephalitis vaccine. J Infect Dis 198: 493499.[Crossref] [Google Scholar]
  32. Lyons A, Kanesa-Thasan N, Kuschner RA, Eckels KH, Putnak R, Sun W, Burge R, Towle AC, Wilson P, Tauber E, Vaughn DW, , 2007. A Phase 2 study of a purified, inactivated virus vaccine to prevent Japanese encephalitis. Vaccine 25: 34453453.[Crossref] [Google Scholar]
  33. Halstead SB, Shotwell H, Casals J, , 1973. Studies on the pathogenesis of dengue infection in monkeys. I. Clinical laboratory responses to primary infection. J Infect Dis 128: 714.[Crossref] [Google Scholar]
  34. Halstead SB, Shotwell H, Casals J, , 1973. Studies on the pathogenesis of dengue infection in monkeys. II. Clinical laboratory responses to heterologous infection. J Infect Dis 128: 1522.[Crossref] [Google Scholar]
  35. Innis BL, Eckels KH, Kraiselburd E, Dubois DR, Meadors GF, Gubler DJ, Burke DS, Bancroft WH, , 1988. Virulence of a live dengue virus vaccine candidate: a possible new marker of dengue virus attenuation. J Infect Dis 158: 876880.[Crossref] [Google Scholar]
  36. Putnak R, Coller B-A, Voss G, Vaughn DW, Clements D, Peters I, Bignami G, Houng H-S, Chen RC, Barvir DA, Seriwatana J, Cayphas S, Garçon 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: 44424452.[Crossref] [Google Scholar]
  37. Sun W, Nisalak A, Gettayacamin M, Eckels KH, Putnak JR, Vaughn DW, Innis BL, Thomas SJ, Endy TP, , 2006. Protection of rhesus monkeys against dengue virus challenge after tetravalent live attenuated dengue virus vaccination. J Infect Dis 193: 16581665.[Crossref] [Google Scholar]
  38. Simmons M, Porter KR, Hayes CG, Vaughn DW, Putnak R, , 2006. Characterization of antibody responses to combinations of a dengue virus type 2 DNA vaccine and two dengue virus type 2 protein vaccines in rhesus macaques. J Virol 80: 95779585.[Crossref] [Google Scholar]
  39. Eckels KH, Dubois DR, Putnak R, Vaughn DW, Innis BL, Henchal EA, Hoke CH, Jr, 2003. Modification of dengue virus strains by passage in primary dog kidney cells: preparation of candidate vaccines and immunization of monkeys. Am J Trop Med Hyg 69: 1216. [Google Scholar]
  40. Blaney JE, Jr Sathe NS, Hanson CT, Firestone CY, Murphy BR, Whitehead SS, , 2007. Vaccine candidates for dengue virus type 1 (DEN1) generated by replacement of the structural genes of rDEN4 and rDEN4Δ30 with those of DEN1. Virol J 4: 23.[Crossref] [Google Scholar]
  41. Simmons M, Burgess T, Lynch J, Putnak R, , 2010. Protection against dengue virus by non-replicating and live attenuated vaccines used together in a prime boost vaccination strategy. Virology 396: 280288.[Crossref] [Google Scholar]
  42. Fernandez S, Cisney ED, Tikhonov AP, Schweitzer B, Putnak RJ, Simmons M, Ulrich RG, , 2011. Antibody recognition of the dengue virus proteome and implications for development of vaccines. Clin Vaccine Immunol 18: 523532.[Crossref] [Google Scholar]
  43. Garçon N, Van Mechelen M, , 2011. Recent clinical experience with vaccines using MPL- and QS-21-containing adjuvant systems. Expert Rev Vaccines 10: 471486.[Crossref] [Google Scholar]
  44. Garçon N, Vaughn DW, Didierlaurent AM, , 2012. Development and evaluation of AS03, an adjuvant system containing α-tocopherol and squalene in an oil-in-water emulsion. Expert Rev Vaccines 11: 349366.[Crossref] [Google Scholar]
  45. Didierlaurent AM, Morel S, Lockman L, Giannini SL, Bisteau M, Carlsen H, Kielland A, Vosters O, Vanderheyde N, Schiavetti F, Larocque D, Van Mechelen M, Garçon N, , 2009. AS04, an aluminum salt- and TLR4 agonist-based adjuvant system, induces a transient localized innate immune response leading to enhanced adaptive immunity. J Immunol 183: 61866197.[Crossref] [Google Scholar]
  46. Morel S, Didierlaurent A, Bourguignon P, Delhaye S, Baras B, Jacob V, Planty C, Elouahabi A, Harvengt P, Carlsen H, Kielland A, Chomez P, Garçon N, Van Mechelen M, , 2011. Adjuvant system AS03 containing α-tocopherol modulates innate immune response and leads to improved adaptive immunity. Vaccine 29: 24612473.[Crossref] [Google Scholar]
  47. Putnak R, Barvir DA, Burrous JM, Dubois DR, D'Andrea 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: 11761184.[Crossref] [Google Scholar]
  48. Sadon N, Delers A, Jarman RG, Klungthong C, Nisalak A, Gibbons RV, Vassilev V, , 2008. A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples. J Virol Methods 153: 16.[Crossref] [Google Scholar]
  49. Risi G, Frenette L, Langley JM, Li P, Riff D, Sheldon E, Vaughn DW, Fries L, , 2011. Immunological priming induced by a two-dose series of H5N1 influenza antigen, administered alone or in combination with two different formulations of AS03 adjuvant in adults: results of a randomised single heterologous booster dose study at 15 months. Vaccine 29: 64086418.[Crossref] [Google Scholar]
  50. Leroux-Roels I, Roman F, Forgus S, Maes C, De Boever F, Dramé M, Gillard P, van der Most R, Van Mechelen M, Hanon E, Leroux-Roels G, , 2010. Priming with AS03A-adjuvanted H5N1 influenza vaccine improves the kinetics, magnitude and durability of the immune response after a heterologous booster vaccination: an open non-randomised extension of a double-blind randomised primary study. Vaccine 28: 849857.[Crossref] [Google Scholar]
  51. Durier C, Desaint C, Lucht F, Girard PM, Lévy Y, May T, Michelet C, Rami A, Roman F, Delfraissy JF, Aboulker JP, Launay O, , 2013. Long-term immunogenicity of two doses of 2009 A/H1N1v vaccine with and without AS03A adjuvant in HIV-1-infected adults. AIDS 27: 8793.[Crossref] [Google Scholar]
  52. Baras B, Bouveret N, Devaster JM, Fries L, Gillard P, Sänger R, Hanon E, , 2008. A vaccine manufacturer's approach to address medical needs related to seasonal and pandemic influenza viruses. Influenza Other Respir Viruses 2: 251260.[Crossref] [Google Scholar]
  53. Blaney JE, Jr Matro JM, Murphy BR, Whitehead SS, , 2005. Recombinant, live-attenuated tetravalent dengue virus vaccine formulations induce a balanced, broad, and protective neutralizing antibody response against each of the four serotypes in rhesus monkeys. J Virol 79: 55165528.[Crossref] [Google Scholar]
  54. Endy TP, Nisalak A, Chunsuttitwat S, Vaughn DW, Green S, Ennis FA, Rothman AL, Libraty DH, , 2004. Relationship of preexisting dengue virus (DV) neutralizing antibody levels to viremia and severity of disease in a prospective cohort study of DV infection in Thailand. J Infect Dis 189: 9901000.[Crossref] [Google Scholar]
  55. Rothman AL, , 2014. DHIM supporting immunologic investigations and the identification of immune correlates of protection. J Infect Dis 209 (Suppl 2): S61S65.[Crossref] [Google Scholar]
  56. Hombach J, Solomon T, Kurane I, Jacobson J, Wood D, , 2005. Report on a WHO consultation on immunological endpoints for evaluation of new Japanese encephalitis vaccines, WHO, Geneva, 2–3 September, 2004. Vaccine 23: 52055211.[Crossref] [Google Scholar]
  57. Van Gessel Y, Klade CS, Putnak R, Formica A, Krasaesub S, Spruth M, Cena B, Tungtaeng A, Gettayacamin M, Dewasthaly S, , 2011. Correlation of protection against Japanese encephalitis virus and JE vaccine (IXIARO®) induced neutralizing antibody titers. Vaccine 29: 59255931.[Crossref] [Google Scholar]
  58. Konishi E, , 2013. Memory B cells: a proposed new immunological correlate for protective efficacy of Japanese encephalitis vaccine. Expert Rev Vaccines 12: 871873.[Crossref] [Google Scholar]

Data & Media loading...

Supplementary PDF

  • Received : 30 Apr 2014
  • Accepted : 04 Dec 2014
  • Published online : 01 Apr 2015

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