Evaluation of endothelial permeability to study dengue pathogenesis and screen com...
| Grant number: | 13/01690-0 |
| Support type: | Regular Research Grants |
| Duration: | March 01, 2014 - August 31, 2016 |
| Field of knowledge: | Biological Sciences - Microbiology |
| Principal Investigator: | Ester Cerdeira Sabino |
| Grantee: | Ester Cerdeira Sabino |
| Home Institution: | Instituto de Medicina Tropical de São Paulo (IMT). Universidade de São Paulo (USP). São Paulo , SP, Brazil |
| Assoc. researchers: | Camila Malta Romano ; Francielle Tramontini Gomes de Sousa |
Abstract
The bleeding is one of the main signs of complication of dengue virus (DENV) infection, causing dengue hemorrhagic fever (DHF), a severe form of the disease with lethal potential especially in children. The DHF pathogenesis is complicated and multifactorial, involving both viral and host factors. Moreover, the lack of animal models representing satisfactorily the pathophysiology of dengue fever in humans has been limiting the advances in understanding the DHF mechanisms as well as the development of drugs for the DHF treatment. Monocytes are cells responsible for immune response to infection with DENV, producing mediators that interact with endothelia and increasing the vascular leakage in humans. Therefore, the aim of this project is to implement a model for assessing the in vitro vascular permeability using endothelial cells co-cultured with monocytes and infected by DENV in order to study the factors involved in vascular extravasation and to screen compounds that possibly inhibit this process. The endothelial cells will be cultivated on inserts with microporous membranes arranged in culture plates, generating two compartments, apical and basolateral. In this system, the cells infected with DENV can produce cytokines and mediators which will interfere with the in vitro vascular permeability, similarly to the in vivo mechanism. The vascular permeability will be calculated through the addition of fluorescent dextran at the apical site and measurement of its transport to the basolateral compartment. The proposed system will allow us to perform the screening of compounds with therapeutic potential against dengue and to study the interference of different clinical isolates of DENV on vascular permeability and the cytokine production profile in order to better understand the factors that interfere in the pathogenesis of DHF. (AU)