A candidate DNA vaccine expressing dengue virus type 1 pre-membrane and envelope proteins was used to immunize rhesus macaques. Monkeys were immunized intramuscularly (i.m.) or intradermally (i.d.) by three or four 1 mg doses of vaccine, respectively. Monkeys that were inoculated i.m. seroconverted more quickly and had higher antibody levels than those that were inoculated i.d. The sera exhibited virus-neutralizing activity, which declined over time. Four of the eight i.m.-inoculated monkeys were protected completely from developing viraemia when challenged 4 months after the last dose with homologous dengue virus. The other four monkeys had reduced viraemia compared with the control immunized monkeys. The i.d.-inoculated monkeys showed no reduction in viraemia when challenged with the virus. All vaccinated monkeys showed an anamnestic antibody response, indicating that they had established immunological memory. Vaccine-induced antibody had an avidity index similar to that of antibody induced by virus infection; however, no clear correlation was apparent between antibody avidity and virus neutralization titres.


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  1. Ansari, M. Z., Shope, R. E. & Malik, S. (1993). Evaluation of vero cell lysate antigen for the ELISA of flaviviruses. Journal of Clinical Laboratory Analysis 7, 230-237.[CrossRef] [Google Scholar]
  2. Bagarazzi, M. L., Boyer, J. D., Ugen, K. E., Javadian, M. A., Chattergoon, M., Shah, A., Bennett, M., Ciccarelli, R., Carrano, R., Coney, L. & Weiner, D. B. (1998). Safety and immunogenicity of HIV-1 DNA constructs in chimpanzees. Vaccine 16, 1836-1841.[CrossRef] [Google Scholar]
  3. Barrett, A. D. (1997a). Yellow fever vaccines. Biologicals 25, 17-25.[CrossRef] [Google Scholar]
  4. Barrett, A. D. (1997b). Japanese encephalitis and dengue vaccines. Biologicals 25, 27-34.[CrossRef] [Google Scholar]
  5. Bhamarapravati, N. & Yoksan, S. (1989). Study of bivalent dengue vaccine in volunteers. Lancet i, 1077.
  6. Boyer, J. D., Ugen, K. E., Wang, B., Agadjanyan, M., Gilbert, L., Bagarazzi, M. L., Chattergoon, M., Frost, P., Javadian, A., Williams, W. V., Refaeli, Y., Ciccarelli, R. B., McCallus, D., Coney, L. & Weiner, D. B. (1997). Protection of chimpanzees from high-dose heterologous HIV-1 challenge by DNA vaccination. Nature Medicine 3, 526-532.[CrossRef] [Google Scholar]
  7. Bray, M., Zhao, B. T., Markoff, L., Eckels, K. H., Chanock, R. M. & Lai, C. J. (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. [Google Scholar]
  8. Chambers, T. J., Hahn, C. S., Galler, R. & Rice, C. M. (1990). Flavivirus genome organization, expression, and replication. Annual Review of Microbiology 44, 649-688.[CrossRef] [Google Scholar]
  9. Colombage, G., Hall, R., Pavy, M. & Lobigs, M. (1998). DNA-based and alphavirus-vectored immunization with prM and E proteins elicits long-lived and protective immunity against the flavivirus, Murray Valley encephalitis virus.Virology 250, 151-163.[CrossRef] [Google Scholar]
  10. Davis, H. L., McCluskie, M. J., Gerin, J. L. & Purcell, R. H. (1996). DNA vaccine for hepatitis B: evidence for immunogenicity in chimpanzees and comparison with other vaccines. Proceedings of the National Academy of Sciences, USA 93, 7213-7218.[CrossRef] [Google Scholar]
  11. Deubel, V., Kinney, R. M., Esposito, J. J., Cropp, C. B., Vorndam, A. V., Monath, T. P. & Trent, D. W. (1988). Dengue 2 virus envelope protein expressed by a recombinant vaccinia virus fails to protect monkeys against dengue. Journal of General Virology 69, 1921-1929.[CrossRef] [Google Scholar]
  12. Donnelly, J. J., Friedman, A., Martinez, D., Montgomery, D. L., Shiver, J. W., Motzel, S. L., Ulmer, J. B. & Liu, M. A. (1995). Preclinical efficacy of a prototype DNA vaccine: enhanced protection against antigenic drift in influenza virus. Nature Medicine 1, 583-587.[CrossRef] [Google Scholar]
  13. Eckels, K. H., Dubois, D. R., Summers, P. L., Schlesinger, J. J., Shelly, M., Cohen, S., Zhang, Y. M., Lai, C. J., Kurane, I., Rothman, A. and others (1994). Immunization of monkeys with baculovirus–dengue type-4 recombinants containing envelope and nonstructural proteins: evidence of priming and partial protection. American Journal of Tropical Medicine and Hygiene 50, 472–478. [Google Scholar]
  14. Edelman, R., Tacket, C. O., Wasserman, S. S., Vaughn, D. W., Eckels, K. H., Dubois, D. R., Summers, P. L. & Hoke, C. H. (1994). A live attenuated dengue-1 vaccine candidate (45AZ5) passaged in primary dog kidney cell culture is attenuated and immunogenic for humans. Journal of Infectious Diseases 170, 1448-1455.[CrossRef] [Google Scholar]
  15. Falgout, B., Bray, M., Schlesinger, J. J. & 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. [Google Scholar]
  16. Gramzinski, R. A., Maris, D. C., Doolan, D., Charoenvit, Y., Obaldia, N., Rossan, R., Sedegah, M., Wang, R., Hobart, P., Margalith, M. & Hoffman, S. (1997). Malaria DNA vaccines in Aotus monkeys. Vaccine 15, 913-915.[CrossRef] [Google Scholar]
  17. Gramzinski, R. A., Millan, C. L., Obaldia, N., Hoffman, S. L. & Davis, H. L. (1998). Immune response to a hepatitis B DNA vaccine in Aotus monkeys: a comparison of vaccine formulation, route, and method of administration. Molecular Medicine 4, 109-118. [Google Scholar]
  18. Gray, D. (1993). Immunological memory. Annual Review of Immunology 11, 49-77.[CrossRef] [Google Scholar]
  19. Gubler, D. J. & Trent, D. W. (1993). Emergence of epidemic dengue/dengue hemorrhagic fever as a public health problem in the Americas. Infectious Agents and Disease 2, 383-393. [Google Scholar]
  20. Halstead, S. B. (1988). Pathogenesis of dengue: challenges to molecular biology. Science 239, 476-481.[CrossRef] [Google Scholar]
  21. Hartmann, G., Weiner, G. J. & Krieg, A. M. (1999). CpG DNA: a potent signal for growth, activation, and maturation of human dendritic cells. Proceedings of the National Academy of Sciences, USA 96, 9305-9310.[CrossRef] [Google Scholar]
  22. Kliks, S. C., Nimmanitya, S., Nisalak, A. & Burke, D. S. (1988). Evidence that maternal dengue antibodies are important in the development of dengue hemorrhagic fever in infants. American Journal of Tropical Medicine and Hygiene 38, 411-419. [Google Scholar]
  23. Kochel, T., Wu, S. J., 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.[CrossRef] [Google Scholar]
  24. Kochel, T. J., Raviprakash, K., Hayes, C. G., Watts, D. M., Russel, K. L., Gozalo, A. S., Phillips, I. A., Ewing, D. W., Murphy, G. S. & Porter, K. R. (2000). A dengue virus serotype-1 DNA vaccine induces virus neutralizing antibodies and provides protection from viral challenge in Aotus monkeys. Vaccine (in press).
  25. Konishi, E., Yamaoka, M., Khin Sane, W., Kurane, I. & 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. [Google Scholar]
  26. Konishi, E., Yamaoka, M., Khin Sane, W., Kurane, I., Takada, K. & Mason, P. W. (1999). The anamnestic neutralizing antibody response is critical for protection of mice from challenge following vaccination with a plasmid encoding the Japanese encephalitis virus premembrane and envelope genes. Journal of Virology 73, 5527-5534. [Google Scholar]
  27. Kreil, T. R., Maier, E., Fraiss, S., Attakpah, E., Burger, I., Mannhalter, J. W. & Eibl, M. M. (1998). Vaccination against tick-borne encephalitis virus, a flavivirus, prevents disease but not infection, although viremia is undetectable. Vaccine 16, 1083-1086.[CrossRef] [Google Scholar]
  28. Krieg, A. M. (1999). CpG DNA: a novel immunomodulator. Trends in Microbiology 7, 64-65.[CrossRef] [Google Scholar]
  29. Lekutis, C., Shiver, J. W., Liu, M. A. & Letvin, N. L. (1997). HIV-1 env DNA vaccine administered to rhesus monkeys elicits MHC class II-restricted CD4+ T helper cells that secrete IFN-gamma and TNF-alpha. Journal of Immunology 158, 4471-4477. [Google Scholar]
  30. Lodmell, D. L., Ray, N. B., Parnell, M. J., Ewalt, L. C., Hanlon, C. A., Shaddock, J. H., Sanderlin, D. S. & Rupprecht, C. E. (1998). DNA immunization protects nonhuman primates against rabies virus. Nature Medicine 4, 949-952.[CrossRef] [Google Scholar]
  31. McKee, K. T.Jr, Bancroft, W. H., Eckels, K. H., Redfield, R. R., Summers, P. L. & Russell, P. K. (1987). Lack of attenuation of a candidate dengue 1 vaccine (45AZ5) in human volunteers. American Journal of Tropical Medicine and Hygiene 36, 435-442. [Google Scholar]
  32. Maniatis, T., Fritsch, E. F. & Sambrook, J. (1984).Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  33. Marchette, N. J. & Halstead, S. B. (1974). Immunopathogenesis of dengue infection in the rhesus monkey. Transplantation Proceedings 6, 197-201. [Google Scholar]
  34. Mason, P. W., McAda, P. C., Mason, T. L. & Fournier, M. J. (1987). Sequence of the dengue-1 virus genome in the region encoding the three structural proteins and the major nonstructural protein NS1. Virology 161, 262-267.[CrossRef] [Google Scholar]
  35. Mathew, A., Kurane, I., Green, S., Stephens, H. A., Vaughn, D. W., Kalayanarooj, S., Suntayakorn, S., Chandanayingyong, D., Ennis, F. A. & Rothman, A. L. (1998). Predominance of HLA-restricted cytotoxic T-lymphocyte responses to serotype-cross-reactive epitopes on nonstructural proteins following natural secondary dengue virus infection. Journal of Virology 72, 3999-4004. [Google Scholar]
  36. Narita, M., Matsuzono, Y., Takekoshi, Y., Yamada, S., Itakura, O., Kubota, M., Kikuta, H. & Togashi, T. (1998). Analysis of mumps vaccine failure by means of avidity testing for mumps virus-specific immunoglobulin G. Clinical and Diagnostic Laboratory Immunology 5, 799-803. [Google Scholar]
  37. Phillpotts, R. J., Venugopal, K. & Brooks, T. (1996). Immunisation with DNA polynucleotides protects mice against lethal challenge with St Louis encephalitis virus. Archives of Virology 141, 743-749.[CrossRef] [Google Scholar]
  38. Porter, K. R., Kochel, T. J., Wu, S. J., Raviprakash, K., Phillips, I. & Hayes, C. G. (1998). Protective efficacy of a dengue 2 DNA vaccine in mice and the effect of CpG immuno-stimulatory motifs on antibody responses. Archives of Virology 143, 997-1003.[CrossRef] [Google Scholar]
  39. Putnak, R., Barvir, D. A., Burrous, J. M., Dubois, D. R., D’Andrea, V. M., Hoke, C. H., Sadoff, J. C. & Eckels, K. H. (1996). Development of a purified, inactivated, dengue-2 virus vaccine prototype in Vero cells: immunogenicity and protection in mice and rhesus monkeys. Journal of Infectious Diseases 174, 1176-1184.[CrossRef] [Google Scholar]
  40. Raviprakash, K., Kochel, T. J., Ewing, D., Simmons, M., Phillips, I., Hayes, C. G. & Porter, K. R. (2000). Immunogenicity of dengue virus type 1 DNA vaccines expressing truncated and full length envelope protein. Vaccine 18, 2426-2434.[CrossRef] [Google Scholar]
  41. Rothman, A. L. & Ennis, F. A. (1999). Immunopathogenesis of dengue hemorrhagic fever. Virology 257, 1-6.[CrossRef] [Google Scholar]
  42. Russell, P. K., Nisalak, A., Sukhavachana, P. & Vivona, S. (1967). A plaque reduction test for dengue virus neutralizing antibodies. Journal of Immunology 99, 285-290. [Google Scholar]
  43. Sanders, V. M., Uhr, J. W. & Vitetta, E. S. (1987). Antigen-specific memory and virgin B cells differ in their requirements for conjugation to T cells. Cell Immunology 104, 419-425.[CrossRef] [Google Scholar]
  44. Schlesinger, R. W. (1977). Dengue viruses. Virology Monographs 16, 1-132. [Google Scholar]
  45. Schmaljohn, C., Vanderzanden, L., Bray, M., Custer, D., Meyer, B., Li, D., Rossi, C., Fuller, D., Fuller, J., Haynes, J. & Huggins, J. (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. Journal of Virology 71, 9563-9569. [Google Scholar]
  46. Scott, R. M., Nisalak, A., Eckels, K. H., Tingpalapong, M., Harrison, V. R., Gould, D. J., Chapple, F. E. & Russell, P. K. (1980). Dengue-2 vaccine: viremia and immune responses in rhesus monkeys. Infection and Immunity 27, 181-186. [Google Scholar]
  47. Simmons, M., Porter, K. R., Escamilla, J., Graham, R., Watts, D. M., Eckels, K. H. & Hayes, C. G. (1998). Evaluation of recombinant dengue viral envelope B domain protein antigens for the detection of dengue complex-specific antibodies. American Journal of Tropical Medicine and Hygiene 58, 144-151. [Google Scholar]
  48. Wang, B., Boyer, J., Srikantan, V., Coney, L., Carrano, R., Phan, C., Merva, M., Dang, K., Agadjanan, M., Gilbert, L. and others (1993). DNA inoculation induces neutralizing immune responses against human immunodeficiency virus type 1 in mice and nonhuman primates. DNA and Cell Biology 12, 799–805.[CrossRef] [Google Scholar]
  49. Yasutomi, Y., Robinson, H. L., Lu, S., Mustafa, F., Lekutis, C., Arthos, J., Mullins, J. I., Voss, G., Manson, K., Wyand, M. & Letvin, N. L. (1996). Simian immunodeficiency virus-specific cytotoxic T-lymphocyte induction through DNA vaccination of rhesus monkeys. Journal of Virology 70, 678-681. [Google Scholar]

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