title: Preparation and immunogenic properties of a recombinant West Nile subunit vaccine.
abstract: 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-gamma, 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.
authors.affiliation: Hawaii Biotech, Inc., 99-193 Aiea Heights Drive, Aiea, HI 96701, USA. mlieberman@hibiotech.com
authors.forename: Michael M
authors.initials: MM
authors.lastname: Lieberman
authors.forename: David E
authors.initials: DE
authors.lastname: Clements
authors.forename: Steven
authors.initials: S
authors.lastname: Ogata
authors.forename: Gordon
authors.initials: G
authors.lastname: Wang
authors.forename: Gloria
authors.initials: G
authors.lastname: Corpuz
authors.forename: Teri
authors.initials: T
authors.lastname: Wong
authors.forename: Tim
authors.initials: T
authors.lastname: Martyak
authors.forename: Lynne
authors.initials: L
authors.lastname: Gilson
authors.forename: Beth-Ann
authors.initials: BA
authors.lastname: Coller
authors.forename: Julia
authors.initials: J
authors.lastname: Leung
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Watts
authors.forename: Robert B
authors.initials: RB
authors.lastname: Tesh
authors.forename: Marina
authors.initials: M
authors.lastname: Siirin
authors.forename: Amelia
authors.initials: A
authors.lastname: Travassos da Rosa
authors.forename: Tom
authors.initials: T
authors.lastname: Humphreys
authors.forename: Carolyn
authors.initials: C
authors.lastname: Weeks-Levy
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000096:biosynthesis; D000917:Antibody Formation / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007111:Immunity, Cellular; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology*; D014901:West Nile Fever / Q000276:immunology* / Q000517:prevention & control* / Q000821:virology; D014902:West Nile virus / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D016207:Cytokines; D014614:Vaccines, Synthetic; D014765:Viral Vaccines