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Immunization of Flavivirus West Nile Recombinant Envelope Domain III Protein Induced Specific Immune Response and Protection against West Nile Virus Infection¹

Flavivirology Laboratory, Department of Microbiology, National University of Singapore, Singapore

The domain III of the West Nile virus (WNV) envelope glycoprotein (E) was shown to serve as virus attachment domain to the cellular receptor, and neutralizing Abs have been mapped to this specific domain. In this study, domain III of the WNV E protein (WNV E DIII) was expressed as a recombinant protein and its potential as a subunit vaccine candidate was evaluated in BALB/C mice. Immunization of WNV E DIII protein with oligodeoxynucleotides (CpG-DNA) adjuvant by i.p. injection was conducted over a period of 3 wk. The immunized mice generated high titer of WNV-neutralizing Abs. Murine Ab against WNV E DIII protein was also capable of neutralizing Japanese encephalitis virus. The IgG isotypes generated were predominantly IgG2a in the murine sera against the recombinant protein. Splenocyte cultures from the mice coadministrated with WNV E DIII protein and CpG secreted large amounts of IFN- and IL-2 and showed proliferation of T cells in the presence of WNV E DIII protein. Overall, this study highlighted that recombinant WNV E DIII protein delivered in combination with CpG adjuvant to mice generated a Th1 immune response type against WNV and can serve as a potential vaccine to prevent WNV infection.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work is supported by grants from the Biomedical Research Council, Singapore (BMRC/01/1/21/18/003), and the National University of Singapore (R-182-000-055-112). J.-H.J.C. was a Singapore Millennium Foundation Postdoctoral Research Fellow.

² Address correspondence and reprint requests to Mah-Lee Ng, Department of Microbiology, 5 Science Drive 2, National University of Singapore, Singapore 117597. E-mail address: micngml{at}nus.edu.sg

³ Abbreviations used in this paper: WNV, West Nile virus; E, envelope; IPTG, isopropyl -D-thiogalactoside; JEV, Japanese encephalitis virus.

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