Development of a Purified, Inactivated, dengue-2 Virus Vaccine Prototype in Vero Cells: Immunogenicity and Protection in Mice and Rhesus Monkeys

J Infect Dis. 1996 Dec;174(6):1176-84. doi: 10.1093/infdis/174.6.1176.


The feasibility of a purified, inactivated dengue (DEN) vaccine made in Vero cells was explored. A DEN-2 virus candidate was chosen for production of a monotypic, purified, inactivated vaccine (PIV). Virus was harvested from roller bottle culture supernatants, concentrated, and purified on sucrose gradients. The purified virus was inactivated with 0.05% formalin at 22 degrees C. After inactivation, the virus retained its antigenicity and was immunogenic in mice and rhesus monkeys, in which it elicited high titers of DEN-2 virus-neutralizing antibody. Mice were completely protected against challenge with live, virulent virus after receiving two 0.15-microg doses of PIV. Monkeys vaccinated with three doses ranging as low as 0.25 microg demonstrated complete absence or a significant reduction in the number of days of viremia after challenge with homologous virus. These results warrant further testing and development of PIVs for other DEN virus serotypes.

MeSH terms

  • Animals
  • Antibodies, Viral / analysis
  • Blotting, Western
  • Cells, Cultured
  • Centrifugation, Density Gradient / methods
  • Chlorocebus aethiops
  • Dengue / immunology*
  • Dengue / prevention & control*
  • Dengue Virus / growth & development
  • Dengue Virus / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • Formaldehyde / pharmacology
  • Hemagglutination Inhibition Tests
  • Macaca mulatta
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred ICR
  • Neutralization Tests
  • Vaccines, Inactivated / administration & dosage
  • Vaccines, Inactivated / chemical synthesis
  • Vaccines, Inactivated / immunology*
  • Vero Cells
  • Viremia / prevention & control


  • Antibodies, Viral
  • Vaccines, Inactivated
  • Formaldehyde