Journal Article

DNA vaccine candidate expressing dengue-3 virus prM and E proteins elicits neutralizing antibodies and protects mice against lethal challenge

De Paula, Sérgio Oliveira, Lima, Danielle Malta, Gomes-Ruiz, Alessandra Cristina, de Oliveira França, Rafael Freitas, da Fonseca, Benedito Antônio Lopes

2008

Overview

In an effort to develop a suitable DNA vaccine candidate for dengue, using dengue-3 virus (DENV-3) as a prototype, the genes coding for premembrane (prM) and envelope proteins (E) were inserted into an expression plasmid. After selecting recombinant clones containing prM/E genes, protein expression in the cell monolayer was detected by indirect immunofluorescence and immunoprecipitation assays. After selecting three vaccine candidates (pVAC1DEN3, pVAC2DEN3 and pVAC3DEN3), they were analyzed in vivo to determine their ability to induce a DENV-3-specific immune response. After three immunizations, the spleens of the immunized animals were isolated, and the cells were cultivated to measure cytokine levels by ELISA and used for lymphoproliferation assays. All of the animals inoculated with the recombinant clones induced neutralizing antibodies against DENV-3 and produced a T cell proliferation response after specific stimuli. Immunized and control mice were challenged with a lethal dose of DENV-3 and observed in order to assess their survival capability. The groups that presented the best survival rate after the challenge were the animals vaccinated with the pVAC3DEN3 clones, with an 80% survival rate. Thus, these data show that we have manufactured a vaccine candidate for DENV-3 that is able to induce a specific immune response and protects mice against a lethal challenge.

• Publisher: Vienna : Springer Vienna,Springer Vienna,Springer,Springer Nature B.V
• Subject: Animals / Antibodies / Antibodies, Viral / Antibodies, Viral - blood / Biological and medical sciences / Biomedical and Life Sciences / Biomedicine / blood / Cell culture / Cell Line, Tumor / Cell Proliferation / Cercopithecus aethiops / Chlorocebus aethiops / clones / Cloning / Cytokines / Cytokines - immunology / Cytokines - metabolism / dengue / Dengue - immunology / Dengue - mortality / Dengue - prevention & control / Dengue fever / Dengue Vaccines / Dengue Vaccines - genetics / Dengue Vaccines - immunology / Dengue Vaccines - therapeutic use / Dengue Virus / Dengue Virus - genetics / Dengue Virus - immunology / Dengue virus type 3 / enzyme-linked immunosorbent assay / Female / Fundamental and applied biological sciences. Psychology / genes / genetics / Genomes / HeLa Cells / Humans / immune response / immunology / Infections / Infectious Diseases / lethal dose / Medical Microbiology / metabolism / Mice / Mice, Inbred BALB C / Microbiology / Miscellaneous / mortality / neutralization / Neutralization Tests / neutralizing antibodies / Original Article / Plasmids / prevention & control / protein synthesis / Proteins / Recombinant Proteins / Recombinant Proteins - genetics / Recombinant Proteins - immunology / recombinant vaccines / survival rate / T-Lymphocytes / T-Lymphocytes - immunology / T-Lymphocytes - metabolism / therapeutic use / Vaccines / Vaccines, DNA / Vaccines, DNA - genetics / Vaccines, DNA - immunology / Vaccines, DNA - therapeutic use / Vero Cells / Viral Envelope Proteins / Viral Envelope Proteins - genetics / Viral Envelope Proteins - immunology / Viral Proteins / Viral Proteins - genetics / Viral Proteins - immunology / Virology / Viruses