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Analysis of the migratory ability of dendritic cells in experimental chromoblastomycosis

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Author(s):
Telma Fátima Emídio Kimura
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
Defense date:
Examining board members:
Sandro Rogerio de Almeida; Tânia Sueli de Andrade; Patrícia Xander Batista
Advisor: Sandro Rogerio de Almeida
Abstract

The chromoblastomycosis is a subcutaneous mycosis with a high morbidity rate, Fonsecaea pedrosoi (F. Pedrosoi) being the largest etiologic agent of this mycosis, featuring a chronic disease, usually confined to the skin and subcutaneous tissues. Rarely do people have cure for this disease, because the therapies shown to be deficient contemporary and few studies report the host-parasite relationship. Dendritic cells (DCs) are specialized in presenting antigens to naïve T lymphocytes inducing primary immune responses. Therefore, we propose to study the migratory capacity of DCs after infection with conidia of F. pedrosoi, since the migration of these cells is intimately linked to its function on T cells, leading to development of a protective adaptive immune response. The phenotype of DCs was evaluated using cells obtained from popliteal lymph nodes, sub cutaneous tissue of BALB/c mice after 12, 24 and 72 hours of infection with conidia of the fungus. Cells were labeled with specific antibodies and analyzed by flow cytometry. After 24 and 72 hours of infection we found a significant decrease in the percentage of DCs in the sub cutaneous tissue, and a significant increase of these cells in the lymph nodes after 72 hours. The expression of surface markers such as CCR7 and costimulatory molecules, were reduced in cells obtained from the sub cutaneous tissue. Like the process of immunophenotyping we can analyze DCs coming from various locations, to better assess the migratory capacity of DCs, cells were stained paws in vivo by injecting dye subcutaneously with CFSE conidia of the fungus. We found that after 12 and 72 hours, DCs sub cutaneous tissue of infected animals migrated to regional lymph nodes. We found that the fungus F. pedrosoi is capable of inducing migration of macrophages and neutrophils to the site of infection within a few hours, leads to increased B-cells in lymph nodes after 12 and 72 hours of infection, and also leads to an increase of CD4 + T lymphocytes in both the site of infection and in the lymph nodes. The results also show that the fungus F. pedrosoi was able to activate the DCs, inducing their migration to regional lymph nodes. (AU)

FAPESP's process: 10/03615-8 - Analysis of the migratory capacity of dendritic cells in chromoblastomycosis experimental
Grantee:Telma Fátima Emidio Kimura
Support Opportunities: Scholarships in Brazil - Master