Inoculation of plasmids expressing the dengue-2 envelope gene elicit neutralizing antibodies in mice.

To develop a nucleic acid vaccine against dengue type-2 virus, the PreM and 92% of the envelope (E) genes were cloned into different eukaryotic plasmid expression vectors (pkCMVint Polyli and pVR1012). The resultant plasmid constructs (pD2ME and P1012D2ME) properly expressed the truncated E protein in vitro as evidenced by the expected protein size on SDS-PAGE and the ability of the protein to be recognized by monoclonal antibodies directed against conformational epitopes. Three-week-old BALB/c mice were given intradermal inoculations of each construct. Plasmid expression vectors without dengue genes were used as controls. One hundred percent of the mice that received the pD2ME and p1012D2ME constructs developed anti-dengue antibodies. These antibodies were shown to neutralize dengue type-2 virus in vitro. This is the first demonstration of the use of nucleic acid inoculation in the development of potential dengue virus. vaccines.