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ALVES, R. P. S. Development of a subunit vaccine against dengue virus serotype 2 based on the non-structural protein 5 (NS5). 2015. 69 p. Master thesis (Microbiology) – Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2015. Dengue fever is a disease caused by four dengue virus serotypes (DENV 1-4), affecting millions of people worldwide and causing a significant number of deaths. There are no effective treatments or vaccine approaches capable of preventing such infection. Anti-DENV vaccine strategies based on nonstructural proteins as antigens have been shown to be safer than those based on structural proteins. In addition, anti-DENV cellular immune responses against nonstructural proteins have shown to play a major role in controlling viral replication. The DENV nonstructural protein 5 (NS5), the most conserved protein among the four serotypes, plays a crucial role in viral replication. In this study, we generated a recombinant form of DENV2 NS5 expressed in E. coli in high amounts and with preserved antigenic properties with regard to the native protein. Culture conditions were optimized in order to allow expression of NS5 as a soluble protein. The immunization of Balb/c mice using this protein alone or in combination with poly (I:C) led to increased survival after intracranial challenge with the DENV2 JHA1 strain. The combination of the protein with poly (I:C) emulsified in Montanide 720 led to the activation of NS5-specific CD8+ T lymphocytes. Altogether, the results indicate that the recombinant NS5 protein preserves antigenic determinants of the native protein and may be a useful tool for studies dealing with the DENV's biology, search for anti-viral drugs and vaccine development.

Desenvolvimento de uma vacina de subunidade contra o sorotipo 2 do vírus dengue baseada na proteína não estrutural 5 (NS5).