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Development of a subunit vaccine against the dengue virus serotype 2 based on the non-structural protein NS5

Grant number: 12/21077-9
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): March 01, 2013
Effective date (End): February 28, 2015
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal Investigator:Luis Carlos de Souza Ferreira
Grantee:Rubens Prince dos Santos Alves
Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil


This project's main objective is to develop and characterize a strategy for prophylactic vaccination against dengue virus serotype 2 (DENV2), based on the combined three adjuvant with NS5 protein and administered subcutaneously in a murine model. This will be cloned and expressed the gene encoding the NS5 DENV2 in recombinant strains of Escherichia coli. The NS5 protein will be administered to BALB/c mice in combination with aluminum hydroxide, rich-CpG oligonucleotides (ODNs) or the toxin LTG33D originally produced by strains of E. enterotoxigenic E. coli (ETEC). Cellular and humoral responses anti-NS5 will be monitored by tests aimed at determining circulating antibodies by ELISA and tests directed to the characterization of cellular responses mediated by T lymphocytes of type CD4 + and CD8 +. This will be assessed the profiles of cytokine secretion and cell activation by ELISPOT and ICS (intracellular cytokine labeling). Further assays are performed in a murine model of challenge to determine the protective ability of the vaccine formulations against infection by DENV2. The safety of such formulations will be evaluated from the analysis of RBC, WBC, circulating cytokines, measuring markers of liver function, levels of C-reactive protein and histopathological analysis. The development of a vaccine based on DENV2 NS5 represent an unprecedented contribution to science, which can then be associated with vaccines aimed at controlling other viral serotypes to generate a tetravalent vaccine formulation capable of preventing dengue humans. Furthermore, the data generated from this project will integrate two research projects funded by FAPESP (ref. Case numbers 2011/51761-6 and 2012/50362-3). (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DOS SANTOS ALVES, RBENS PRINCE; PEREIRA, LENNON RAMOS; NASCIMENTO FABRIS, DENICAR LINA; SALVADOR, FELIPE SCASSI; SANTOS, ROBERT ANDREATA; DE ANDRADE ZANOTTO, PAOLO MARINHO; ROMANO, CAMILA MALTA; AMORIM, JAIME HENRIQUE; DE SOUZA FERREIRAA, LUIS CARLOS. Production of a Recombinant Dengue Virus 2 NS5 Protein and Potential Use as a Vaccine Antigen. Clinical and Vaccine Immunology, v. 23, n. 6, p. 460-469, . (15/02352-7, 12/21077-9, 14/17595-0)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
ALVES, Rubens Prince dos Santos. Development of a subunit vaccine against dengue virus serotype 2 based on the non-structural protein 5 (NS5).. 2015. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.

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