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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2016.tde-25082016-151946
Document
Author
Full name
Keyde Cristina Martins de Melo
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Ruiz, Rita de Cássia (President)
Oliveira, Maria Leonor Sarno de
Silva, Rosa Maria
Souza, Ana Olívia de
Yamamoto, Denise
Title in Portuguese
Interação de Escherichia coli enteropatogênica (EPEC) atípica com fagócito profissional.
Keywords in Portuguese
Antifagocitose
EPEC atípica
Fagocitose
Macrófago
Óxido nítrico
Rab5
Rab7
Abstract in Portuguese
O aumento dos casos de diarreia causados por EPECa evoca a sua capacidade de adaptação e patogenicidade. O objetivo deste estudo foi investigar o comportamento da EPECa na interação com macrófagos (fagocitose e antifagocitose). O estudo da fagocitose das cepas LB7 (O55:H7), LB13 (O111:abH9) e BA487 (O55:H7) em macrófagos J774A1 mostrou sobrevivência intracelular em presença de NO. EPECa impede a maturação do vacúolo parasitóforo. A sobrevivência em macrófago derivado de C3H/HeJ, mutante do tlr4, foi reduzida e em macrófago de C57BL/6 não foi observada. O fator antigagocítico (Fa) secretado pela LB7, já detectado pelo grupo, apresenta natureza peptídica e a sua ação não é específica de EPEC, pois inibe também a fagocitose de Shigella e látex. A secreção do Fa foi avaliada em M9 e DMEM. O produto do fracionamento do sobrenadante do cultivo por SPE apresentou Fa em ambos meios. No entanto, a secreção em M9 é baixa e não foi detectada por HPLC. O Fa do DMEM obtido por HPLC mostrou-se citotóxico. Novos meios de cultivo deverão ser estudados para a identificação do Fa.
Title in English
Interaction atypical enteropathogenic Escherichia coli (EPEC) with professional phagocytes.
Keywords in English
Anti-phagocytosis
Atypical EPEC
Macrophage
Nitric Oxide
Phagocytosis
Rab5
Rab7
Abstract in English
The increase in the numbers of diarrhea cases caused by aEPEC denotes its adaptability and pathogenicity. The objective of this study was to investigate the behavior of aEPEC in the interaction with macrophages (phagocytosis and anti-phagocytosis). Strains LB7 (O55:H7), LB13 (O111:abH9) and BA487 (O55:H7) were shown to survive within J774A1 macrophages in the presence of NO. aEPEC prevents maturation of the parasitophorus vacuole. Survival inside C3H/HeJ derived macrophages, mutant for tlr4, decreased and was not observed in C57BL/6 derived macrophages. The anti-phagocytic factor (AF), secreted by LB7 and previously detected by our group, is peptidic and its action is not specific to EPEC as it also inhibits phagocytosis of Shigella and latex. Secretion of AF was evaluated in M9 and DMEM. AF was detected after SPE fractionation of both culture media. However, the secretion in M9 is low and was not detected by HPLC. The DMEM HPLC fraction containing AF was cytotoxic. New culture media will be studied for the identification of AF.
 
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Publishing Date
2016-08-25
 
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