Production of a Recombinant Dengue Virus 2 NS5 Protein and Potential Use as a Vaccine Antigen

Author(s)

dos Santos Alves, Rbens Prince ^[1] ; Pereira, Lennon Ramos ^[1] ; Nascimento Fabris, Denicar Lina ^[1] ; Salvador, Felipe Scassi ^[2] ; Santos, Robert Andreata ^[1] ; de Andrade Zanotto, Paolo Marinho ^[3] ; Romano, Camila Malta ^[2] ; Amorim, Jaime Henrique ^{[1, 4]} ; de Souza Ferreiraa, Luis Carlos ^[1]

Affiliation

^[1] Univ Sao Paulo, Dept Microbiol, Inst Ciencias Biomed, Lab Desenvolvimento Vacinas, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Escola Med, Inst Trop Med, Sao Paulo - Brazil

^[3] Univ Sao Paulo, Dept Microbiol, Inst Ciencias Biomed, Lab Evolucao & Mol Bioinformat, Sao Paulo - Brazil

^[4] Univ Fed Oeste Bahia, Ctr Ciencias Biol & Saude, Barreiras, BA - Brazil

• Document type: Journal article
• Source: Clinical and Vaccine Immunology; v. 23, n. 6, p. 460-469, JUN 2016.
• Web of Science Citations: 4

Abstract

Dengue fever is caused by any of the four known dengue virus serotypes (DENV1 to DENV4) that affect millions of people worldwide, causing a significant number of deaths. There are vaccines based on chimeric viruses, but they still are not in clinical use. Anti-DENV vaccine strategies based on nonstructural proteins are promising alternatives to those based on whole virus or structural proteins. The DENV nonstructural protein 5 (NS5) is the main target of anti-DENV T cell-based immune responses in humans. In this study, we purified a soluble recombinant form of DENV2 NS5 expressed in Escherichia coli at large amounts and high purity after optimization of expression conditions and purification steps. The purified DENV2 NS5 was recognized by serum from DENV1-, DENV2-, DENV3-, or DENV4-infected patients in an epitope-conformation-dependent manner. In addition, immunization of BALB/c mice with NS5 induced high levels of NS5-specific antibodies and expansion of gamma interferon-and tumor necrosis factor alpha-producing T cells. Moreover, mice immunized with purified NS5 were partially protected from lethal challenges with the DENV2 NGC strain and with a clinical isolate (JHA1). These results indicate that the recombinant NS5 protein preserves immunological determinants of the native protein and is a promising vaccine antigen capable of inducing protective immune responses. (AU)

• FAPESP's process: 15/02352-7 - Vaccines for control of dengue: vaccine potential of the combination of the non-structural proteins in the generation of protective cell response in an experimental model
• Grantee: Rúbens Prince dos Santos Alves
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 12/21077-9 - Development of a subunit vaccine against the dengue virus serotype 2 based on the non-structural protein NS5
• Grantee: Rúbens Prince dos Santos Alves
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 14/17595-0 - New vaccine research based on recombinant proteins of dengue virus
• Grantee: Luis Carlos de Souza Ferreira
• Support Opportunities: Regular Research Grants