| Congresso Brasileiro de Microbiologia 2023 | Resumo: 736-1 | ||||
Resumo:The Zika virus (ZIKV) is an arbovirus that has a great impact on global health and has drawn attention due to the pathogenesis associated with infections and the rapid dissemination in many countries, especially tropical ones with vector mosquitoes, such as Brazil. Therefore, continued investment in programs to control and prevent is essential. The goal of this study is to develop an effective vaccine against ZIKV, using VLPs platforms as a delivery of the ED-III antigen. VLPs (Qβ) and ED-III ZIKV proteins were obtained in prokaryotic expression systems, through in a bacterial platform (Escherichia coli BL21), purified by chromatography, and analyzed by electrophoresis. After vaccine formulation, in which the ED-III protein was chemically conjugated to QβVLPs, isogenic C57BL/6 mice were immunized with two doses intramuscularly, and blood samples were collected weekly by submandibular venous plexus puncture to assess the humoral and cellular immune response. At the end of 42 days, the spleen was collected and processed for analysis by flow cytometry. The production of antibodies was quantified using the enzyme-linked immunosorbent assay technique. Statistical analyses were performed using the R language. At the end of the protein purification was obtained approximately 3 mg of pure ED-III protein per 1L of culture. The vaccine formulation was analyzed by SDS-PAGE, Western blot and transmission electron microscopy. Flow cytometry analysis revealed a statistical significance increase of intracellular cytokine (IFN-γ and TNF-α) compared to the control groups. In the humoral immune response, the production of total IgG antibodies and subclasses were evidenced, indicating elevated levels of these molecules in the vaccine groups. Therefore, both the protein and VLP were produced, purified, and validated regarding the preservation of antigenicity and functionality. They exhibited high quality for application in vaccine antigen assays. The vaccine formulations demonstrated immunogenicity, inducing high titers of antigen-specific antibodies in an animal model, and conferring a cellular immune response, which is considered crucial for generating protective responses against ZIKV infection. Palavras-chave: Arboviruses, Protein expression, Vaccine, VLPs, Zika virus Agência de fomento:FAPESP | |||||