Preparation and immunogenic properties of a recombinant West Nile subunit vaccine

Michael M. Lieberman, David E. Clements, Steven Ogata, Gordon Wang, Gloria Corpuz, Teri Wong, Tim Martyak, Lynne Gilson, Beth Ann Coller, Julia Leung, Douglas M. Watts, Robert B. Tesh, Marina Siirin, Amelia Travassos da Rosa, Tom Humphreys, Carolyn Weeks-Levy

Research output: Contribution to journalArticlepeer-review

60 Scopus citations


While several West Nile vaccines are being developed, none are yet available for humans. In this study aimed at developing a vaccine for humans, West Nile virus (WNV) envelope protein (E) and non-structural protein 1 (NS1) were produced in the Drosophila S2 cell expression system. The C-terminal 20% of the E protein, which contains the membrane anchor portion, was deleted, thus allowing for efficient secretion of the truncated protein (80E) into the cell culture medium. The proteins were purified by immunoaffinity chromatography (IAC) using monoclonal antibodies that were flavivirus envelope protein group specific (for the 80E) or flavivirus NS1 group specific (for NS1). The purified proteins were produced in high yield and used in conjunction with adjuvant formulations to vaccinate mice. The mice were tested for both humoral and cellular immune responses by a plaque reduction neutralization test and ELISA, and by lymphocyte proliferation and cytokine production assays, respectively. The results revealed that the 80E and the NS1 proteins induced both high-titered ELISA and neutralizing antibodies in mice. Splenocytes from immunized mice, cultured in vitro with the vaccine antigens as stimulants, showed excellent proliferation and production of cytokines (IFN-γ, IL-4, IL-5, and IL-10). The level of antigen-stimulated lymphocyte proliferation and cytokine production was comparable to the level obtained from mitogen (phytohemagglutinin or pokeweed) stimulation, indicating a robust cellular response as well. These findings are encouraging and warrant further in vivo studies to determine the protective efficacy of the WNV vaccine candidate.

Original languageEnglish (US)
Pages (from-to)414-423
Number of pages10
Issue number3
StatePublished - Jan 5 2007


  • Recombinant subunit vaccine
  • West Nile vaccine

ASJC Scopus subject areas

  • Molecular Medicine
  • General Immunology and Microbiology
  • General Veterinary
  • Public Health, Environmental and Occupational Health
  • Infectious Diseases


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