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

Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Clinical and Vaccine Immunology; v. 23, n. 6, p. 460-469, JUN 2016.
• Citações Web of Science: 4

Resumo

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)

• Processo FAPESP: 12/21077-9 - Desenvolvimento de uma vacina de subunidade contra o sorotipo 2 do vírus dengue baseada na proteína não estrutural 5 (NS5).
• Beneficiário: Rubens Prince dos Santos Alves
• Linha de fomento: Bolsas no Brasil - Mestrado
• Processo FAPESP: 14/17595-0 - Novas pesquisas vacinais baseadas em proteínas recombinantes do vírus dengue
• Beneficiário: Luis Carlos de Souza Ferreira
• Linha de fomento: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/02352-7 - Vacinas para o controle da dengue: potencial vacinal da combinação das proteínas não estruturais na geração de resposta celular protetora em modelo experimental
• Beneficiário: Rubens Prince dos Santos Alves
• Linha de fomento: Bolsas no Brasil - Doutorado