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Master's Dissertation
DOI
https://doi.org/10.11606/D.9.2012.tde-01032013-092230
Document
Author
Full name
Telma Fátima Emídio Kimura
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Almeida, Sandro Rogerio de (President)
Andrade, Tânia Sueli de
Batista, Patrícia Xander
Title in Portuguese
Análise da capacidade migratória de células dendríticas na cromoblastomicose experimental
Keywords in Portuguese
Células dendríticas
Cromoblastomicose (Medicina)
Imunologia
Micologia médica
Migração celular
Abstract in Portuguese
A cromoblastomicose é uma micose subcutânea, com alto índice de morbidade, sendo Fonsecaea pedrosoi (F. pedrosoi) considerado o maior agente etiológico dessa micose, caracterizando uma doença crônica, geralmente confinada na pele e tecidos subcutâneos. Raramente os indivíduos apresentam cura dessa doença, pois as terapias contemporâneas mostram-se deficientes e poucos trabalhos relatam a relação parasito-hospedeiro. As células dendríticas (DCs) são especializadas na apresentação de antígenos para linfócitos T naive induzindo respostas imunes primárias. Diante disso, propomos estudar a capacidade migratória de DCs após infecção com conídios de F. pedrosoi, uma vez que o processo de migração dessas células está intimamente ligado com a sua função sobre as células T, levando ao desenvolvimento de uma resposta imune adaptativa protetora. O fenótipo de DCs foi avaliado através de células obtidas dos linfonodos poplíteos, inguinais e patas de camundongos BALB/c após 12, 24 e 72 horas de infecção com conídios do fungo. Células obtidas foram marcadas com anticorpos específicos e analisadas por citometria de fluxo. Após 24 e 72 horas de infecção verificamos uma diminuição significativa na porcentagem de DCs nas patas, e um aumento significativo dessas células nos linfonodos após 72 horas. A expressão de marcadores de superfície como CCR7 e moléculas co-estimulatórias, mostraram-se diminuídas nas células obtidas das patas. Como no processo de imunofenotipagem podemos analisar DCs vindas de diversos locais, para melhor avaliar a capacidade migratória das DCs, células das patas foram marcadas in vivo injetando-se subcutaneamente o corante CFSE juntamente com conídios do fungo. Verificamos que após 12 e 72 horas, as DCs das patas dos animais infectados, migraram para os linfonodos regionais. Assim constatamos que o fungo F. pedrosoi é capaz de induzir a migração de macrófagos e neutrófilos para o sítio de infecção em poucas horas, leva ao aumento de células B nos linfonodos após 12 e 72 horas de infecção, e também leva ao aumento de linfócitos T CD4+ tanto no local de infecção quanto nos linfonodos. Os resultados também comprovam que o fungo F. pedrosoi foi capaz de ativar as DCs, induzindo sua migração para os linfonodos regionais.
Title in English
Analysis of the migratory ability of dendritic cells in experimental chromoblastomycosis
Keywords in English
Cell migration
Chromoblastomycosis (Medicine)
Dendritic cells
Immunology
Medical micology
Abstract in English
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.
 
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Publishing Date
2013-04-08
 
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