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Development of a subunit vaccine against the dengue virus serotype 2 based on the protein NS3 helicase domain.

Grant number: 11/14212-4
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): May 01, 2012
Effective date (End): April 30, 2014
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Luis Carlos de Souza Ferreira
Grantee:Raiza Sales Pereira Bizerra
Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil


This project aims to develop and characterize a strategy for prophylactic vaccination against dengue virus serotype 2 (DENV2), based on NS3 protein helicase domain (NS3H) combined with three adjuvants and injected subcutaneously in mice. To do this, will be cloned and expressed the genes encoding NS3H and a non-toxic heat-labile toxin (LTG33D) from enterotoxigenic E. coli (ETEC) in recombinant strains of E. coli. Protein NS3H will be administered to BALB/c mice in combination with aluminum hydroxide, CpG-rich oligonucleotides (ODNs) or LTG33D. The cellular and humoral responses against NS3H will be monitored by testing the determination of circulating antibodies by ELISA assays and characterizing cellular responses mediated by T lymphocytes CD4+ and CD8+. The profiles of cytokine secretion and cell activation will be assessed by ELISPOT and ICS (intracellular cytokine staining). Further tests will be performed in a murine model challenge to determine the protective ability of vaccine formulations against the DENV2 infection. The safety of such formulations will be evaluated by analysing red and white blood cells counts, circulating cytokines, markers of liver function, levels of C-reactive protein and by pathological examinations. The development of a DENV2 vaccine represent an unprecedented contribution to science, which may later be associated with vaccines for other viral serotypes in order to generate a tetravalent vaccine formulation able to prevent dengue fever in humans. Furthermore, the data generated from this project will represent an important contribution to studies aimed at the development of vaccination strategies against dengue.

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Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
BIZERRA, Raiza Sales Pereira. Development of a subunit vaccine against dengue virus serotype 2 based on the NS3 helicase domain.. 2014. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.

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