Development and evaluation of an enzyme-linked immunosorbent assay for quantifying antibodies to Japanese encephalitis virus nonstructural 1 protein to detect subclinical infections in vaccinated horses.

Antibodies to Japanese encephalitis virus (JEV) nonstructural 1 (NS1) protein constitute a marker of natural JEV infection among populations vaccinated with inactivated JE vaccine. In Japan, with few recent human JE cases, the natural infection rate is critical to evaluate the necessity of continuing JE vaccination. A sensitive immunochemical staining method for detecting NS1 antibodies in individuals naturally and subclinically infected with JEV was previously established.

Comparison of protective efficacies of plasmid DNAs encoding Japanese encephalitis virus proteins that induce neutralizing antibody or cytotoxic T lymphocytes in mice.

Mice immunized with a plasmid DNA encoding the premembrane (prM) and envelope (E) proteins of Japanese encephalitis (JE) virus (designated pcJEME) produce neutralizing antibodies and are protected from JE. To determine the role of the immune response to other viral proteins in protection, we constructed plasmid DNAs encoding other JE virus proteins and made a direct comparison among these plasmids using a mouse model. Cytotoxic T lymphocytes (CTLs) were induced by plasmids encoding capsid (C) or nonstructural proteins, NS1, NS2A, NS2B, NS3 or NS5.

Enhancing effect of vaxfectin on the ability of a Japanese encephalitis DNA vaccine to induce neutralizing antibody in mice.

Vaxfectin, a recently developed adjuvant, was evaluated for its enhancing effect on immunogenicity of a Japanese encephalitis (JE) DNA vaccine plasmid encoding the JE virus premembrane (prM) and envelope (E) genes (designated pcJEME), using BALB/c and ICR mice. Formulation of pcJEME with Vaxfectin provided > or =8-fold higher neutralizing antibody titers than those induced by pcJEME alone and reduced the amount of pcJEME to one-tenth to induce comparable levels of neutralizing antibody. Use of Vaxfectin did not alter a Th1 type IgG isotype immune response (IgG1 < IgG2a) induced by pcJEME in mice.