Development of Safe, Effective Vaccines for Dengue Virus Disease by RecombinantBaculovirus. Subtitle: Development of Safe, Effective Vaccines for Dengue Disease Utilizing Viral Envelope and NSI Glycoproteins Expressed by Recombinant Baculovirus

摘要

During the initial phase of this study we have completed construction ofrecombinant baculoviruses that separately express full-length E or NS1 glycoprotein of dengue type 2 and type 4 viruses. Radio-immunoprecipitation was performed to analyze the dengue product in the lysate of recombinant virus infected cells. The results showed that insect cells infected with recombinant b (DEN 4, 100%E) produced a protein with molecular weight of 55-60 kilodalton (Kd) as predicted for the glycosylated form of dengue E glycoprotein. Similarly, recombinant b (DEN 4, NS1) or b (DEN 2, NS1) produced a protein in infected insect cells that was approximately 40-44 Kd, the size predicted for the dengue NS1 glycoprotein. In the next phase of this study we plan to prepare lysates of insect cells infected with various recombinant baculoviruses. These lysates will be analyzed quantitatively and used for evaluation of immunogenicity of the expressed dengue protein products in mice. These studies will allow us to determine separately the protective efficacy afforded by each of these dengue virus proteins, and therefore, select the most immunogenic as well as the most protective recombinant products for further evaluation in primates.