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: Rúbens Prince dos Santos Alves
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

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)

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, JUN 2016. (15/02352-7, 12/21077-9, 14/17595-0)

ALVES, RUBENS PRINCE DOS SANTOS; ANDREATA-SANTOS, ROBERT; DE FREITAS, CARLA LONGO; PEREIRA, LENNON RAMOS; FABRIS-MAEDA, DENICAR LINA NASCIMENTO; RODRIGUES-JESUS, MONICA JOSIANE; PEREIRA, SAMUEL SANTOS; CARVALHO, ALEXIA ADRIANNE VENCESLAU BRITO; SALES, NATIELY SILVA; PERON, JEAN PIERRE SCHATZMANN; et al. Protective Immunity to Dengue Virus Induced by DNA Vaccines Encoding Nonstructural Proteins in a Lethal Challenge Immunocompetent Mouse Model. FRONTIERS IN MEDICAL TECHNOLOGY, v. 2, p. 12-pg., 2020-10-30. (17/26170-0, 14/17595-0, 15/02352-7, 12/21077-9)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

ALVES, Rúbens 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.