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Phenotypic and transcriptome evaluation of murine and human leukocytes in Leishmania infantum infection

Grant number: 16/18357-0
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): December 01, 2016
Effective date (End): August 31, 2017
Field of knowledge:Biological Sciences - Immunology - Cellular Immunology
Principal Investigator:João Santana da Silva
Grantee:Bruna Araújo David
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:13/08216-2 - CRID - Center for Research in Inflammatory Diseases, AP.CEPID


Visceral Leishmaniasis (VL) is a severe chronic disease, and it is potentially lethal in humans, whose lethality rate can reach 10% when not establishing proper treatment. The main goal of this project is to perform immunophenotypic comparison and transcriptome analysis of leukocytes aiming to identify genes that are differentially expressed during the disease. We also aim to identify the cellular processes that may be related to the different events during human infection with L. infantum, and the development of Visceral Leishmaniasis (VL). For this murine and human leukocytes will be immunophenotyped by flow cytometry combined with mass spectrometry (CyTOF, Time-of-flight mass cytometry) in order to determine the main cellular populations involved in the infectious process as well as phenotypic alterations of these cells. The transcriptome study will be done by individual sequencing of cell populations of interest selected from the results of CyTOF. Moreover, with the data obtained from the RNA sequencing, relevant cellular pathways during Visceral Leishmaniasis as well as participation of inflamassomes and cytokine profile throughout the infectious process may be revealed. Validation of the results of the RNA sequencing will be made using gene silencing techniques, conditional knockout mice strains and depletion of selective blockage of different cytokines. This would also impact on the elucidation of novel therapeutic strategies. Finally, the dynamics of leukocyte recruitment in a murine model of Visceral Leishmaniasis will be performed using confocal intravital microscopy. The execution of this project will contribute significantly to our understanding of cell populations and mechanisms involved in infection by L. infantum with great potential to establish intervention mechanisms in the infectious process leading to optimize the treatment of patients and also assist in the disease diagnosis. (AU)

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