Live-attenuated dengue vaccine development
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645


Laboratory-attenuated strains of each of the four dengue serotypes previously tested as monovalent vaccines in volunteers were combined and tested for immunogenicity, safety, and reactogenicity in 16 dosage combinations. Tetravalent vaccines made using combinations of high (10 plaque-forming units [PFU]/dose) or low (10 PFU/dose) dosage formulations of each of the four viruses were inoculated in 64 flavivirus non-immune adult volunteers to determine which, if any, formulation raised neutralizing antibodies in at least 75% of volunteers to at least three of four dengue serotypes following one or two inoculations. Such formulations, if safe and sufficiently non-reactogenic, would be considered for an expanded Phase II trial in the future. Formulations 1–15 were each inoculated into three or four volunteers (total = 54) on days 0 and 28. Formulation 16 was tested in 10 volunteers, five volunteers inoculated on days 0 and 30, one volunteer on days 0 and 120, and four volunteers on days 0, 30, and 120. Blood was drawn for serologic assays immediately before and one month after each vaccination, and for viremia assay on day 10 after each vaccination. The 16 formulations were safe, but variably reactogenic after the first vaccination, and nearly non-reactogenic after the second and third vaccinations. Reactogenicity was positively correlated with immunogenicity. Similar proportions of volunteers seroconverted to dengue-1 (69%), dengue-2 (78%), and dengue-3 (69%), but significantly fewer volunteers seroconverted to dengue-4 (38%). The geometric mean 50% plaque reduction neutralization test titers in persons who seroconverted were significantly higher to dengue-1 (1:94) than to dengue-2 (1:15), dengue-3 (1:10), and dengue-4 (1:2). Seven formulations met the serologic criteria required for an expanded trial, and three of these were sufficiently attenuated clinically to justify further testing.


Article metrics loading...

Loading full text...

Full text loading...



  1. Brandt WE, 1990. From the World Health Organization. Development of dengue and Japanese encephalitis vaccines. J Infect Dis 162 : 577–583.
  2. Halstead SB, Palumbo NE, 1973. Studies on the immunization of monkeys against dengue. II. Protection following inoculation of combinations of viruses. Am J Trop MedHyg 22 : 375–381.
  3. Edelman R, Tacket CO, Wasserman SS, Vaughn DW, Eckels KH, Dubois DR, Summers PL, Hoke CH, 1994. A live attenuated dengue-1 vaccine candidate (45AZ5) passaged in primary dog kidney cell culture is attenuated and immunogenic for humans. J Infect Dis 170 : 1448–1455.
  4. Kanesa-thasan N, Edelman R, Tacket CO, Wasserman SS, Vaughn DW, Coster TS, Kim-Ahn D, Dubois DR, Putnak JR, King A, Summers PL, Innis BL, Eckels KH, Hoke CH Jr, 2003. 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 (suppl): (in press).
  5. Sun W, Edelman R, Kanesa-thasan N, Eckels KH, Putnak R, King AD, Huong H-S, 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 (suppl): (in press).
  6. Sutter RW, Cochi SL, Melnick JL, 1999. Live attenuated polio vaccines. Plotkin SA, Orenstein WA, eds. Vaccines. Philadelphia: W. B. Saunders, 364–408.
  7. McKee KT Jr, Bancroft WH, Eckels KH, Redfield RR, Summers PL, Russell PK, 1987. Lack of attenuation of a candidate dengue 1 vaccine (45AZ5) in human volunteers. Am J Trop Med Hyg 36 : 435–442.
  8. Eckels KH, Scott RM, Bancroft WH, Brown J, Dubois DR, Summers PL, Russell PK, Halstead SB, 1984. Selection of attenuated dengue 4 viruses by serial passage in primary kidney cells. V. Human response to immunization with a candidate vaccine prepared in fetal rhesus lung cells. Am J Trop Med Hyg 33 : 684–689.
  9. Russell PK, Nisalak A, Sukhavachana P, Vivona S, 1967. A plaque reduction test for dengue virus neutralizing antibodies. J Immunol 99 : 285–290.
  10. Houng HSH, Chen RCM, Vaughn DW, Kanesa-Thasan N, 2001. Development of a fluorogenic RT-PCR system for quantitative identification of dengue virus serotypes 1–4 using conserved and serotype-specific 3′ noncoding sequences. J Virol Methods 95 : 19–32.
  11. Sabin A, 1952. Research on dengue in World War II. Am J Trop Med Hyg 1 : 30–50.
  12. Kanesa-thasan N, Sun W, Kim-Ahn G, Van Albert S, Putnak JR, King A, Raengsakulsrach B, Christ-Schmidt H, Gilson K, Zahradnik JM, Vaughn DW, Innis BL, Saluzzo J, Hoke CH, 2001. Safety and immunogenicity of attenuated dengue virus vaccines (Aventis Pasteur) in human volunteers. Vaccine 19 : 3179–3188.
  13. Bravo JR, Guzman MG, Kouri GP, 1987. Why dengue haemorrhagic fever in Cuba? 1. Individual risk factors for dengue haemorrhagic fever/dengue shock syndrome (DHF/DSS). Trans R Soc Trop Med Hyg 81 : 816–820.
  14. Bancroft WH, Top FH, Eckels KH, Anderson JH, McCown JM, Russell PK, 1981. Dengue-2 vaccine: virological, immunological, and clinical responses of six yellow fever-immune recipients. Infect Immun 31 : 698–703.
  15. Scott RM, Eckels KH, Bancroft WH, Summers PL, McCown JM, Anderson JH, Russell PK, 1983. Dengue 2 vaccine: dose response in volunteers in relation to yellow fever immune status. J Infect Dis 148 : 1055–1060.
  16. Bancroft WH, Scott RM, Eckels KH, Hoke CH, Simms TE, Jesrani KD, Summers PL, Dubois DR, Tsoulos D, Russell PK, 1984. Dengue virus type 2 vaccine: reactogenicity and immunogenicity in soldiers. J Infect Dis 149 : 1005–1010.
  17. 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 : 876–880.
  18. Hoke CH, Malinoski FJ, Eckels KH, Scott RM, Dubois DR, Summers PL, Simms T, Burrous J, Hasty SE, Bancroft WH, 1990. Preparation of an attenuated dengue 4 (341750 Carib) virus vaccine. II. Safety and immunogenicity in humans. Am J Trop Med Hyg 43 : 219–226.
  19. Bhamarapravati N, Yoksan S, 1989. Study of bivalent dengue vaccine in volunteers (letter). Lancet 1 : 1077.
  20. Bhamarapravati N, Yoksan S, Chayaniyayothin T, Angsubphakorn S, Bunyaratvej A, 1987. Immunization with a live attenuated dengue-2-virus candidate vaccine (16681-PDK 53): clinical, immunological and biological responses in adult volunteers. Bull World Health Organ 65 : 189–195.
  21. Bhamarapravati N, Sutee Y, 2000. Live attenuated tetravalent dengue vaccine. Vaccine 18 (suppl 2): 44–47.
  22. Vaughn DW, Hoke CH Jr, Yoksan S, LaChance R, Innis BL, Rice RM, Bhamarapravati N, 1996. Testing of a dengue 2 live-attenuated vaccine (strain 16681 PDK 53) in ten American volunteers. Vaccine 14 : 329–336.
  23. Henchal EA, Henchal LS, Schlesinger JJ, 1988. Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue 2 virus. J Gen Virol 69 : 2101–2107.
  24. Kaufman BM, Summers PL, Dubois DR, Cohen WH, Gentry MK, Timchak RL, Burke DS, Eckels KH, 1989. Monoclonal antibodies for dengue virus prM glycoprotein protect mice against lethal dengue infection. Am J Trop Med Hyg 41 : 576–580.
  25. Kliks SC, Nisalak A, Brandt WE, Wahl L, Burke DS, 1989. Antibody-dependent enhancement of dengue virus growth in human monocytes as a risk factor for dengue hemorrhagic fever. Am J Trop Med Hyg 40 : 444–451.

Data & Media loading...

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