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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2010.tde-05102010-154953
Document
Author
Full name
Sérgio Paulo Dejato da Rocha
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Elias Junior, Waldir Pereira (President)
Amaral, Tania Aparecida Tardelli Gomes do
Carbonare, Solange Barros
Ferreira, Luis Carlos de Souza
Oliveira, Maria Leonor Sarno de
Title in Portuguese
Análise estrutural e funcional da região LEE de Escherichia coli enteropatogênica atípica.
Keywords in Portuguese
Escherichia coli
Diarréia
Interação celular
Abstract in Portuguese
aEPEC é capaz de causar lesão A/E, provocada por proteínas codificadas na região LEE. Foi realizada a análise estrutural e funcional da região LEE de amostras de aEPEC que expressam os padrões ALL, AA e AD, e amostra não aderente (NA). O padrão de adesão característico e capacidade de causar a lesão A/E foram investigados em células epiteliais. As amostras mantiveram o padrão de adesão independentemente da origem da linhagem celular. A lesão A/E foi detectada em algumas linhagens celulares após o contato com as amostras ALL e AD. A presença da região LEE foi detectada intacta e ensaios de PCR em tempo real, microarray e imunodetecção, mostrando a funcionalidade da mesma em todas as amostras. Um plasmídio que expressa a proteína EspFu foi introduzido em todas as 4 amostras, demonstrando não influenciar nos padrões de adesão e nem na capacidade de causar a lesão A/E nas amostras ALL, AA e AD. Mas, a amostra NA expressou o padrão ALL e foi capaz de causar a lesão A/E. Assim, EspFu desempenhou papel na adesão celular além do estabelecimento da lesão A/E in vitro.
Title in English
Structural and functional analysis of LEE region of atypical enteropathogenic Escherichia coli.
Keywords in English
Escherichia coli
Cellular interaction
Diarrhea
Abstract in English
aEPEC is capable to cause A/E lesion, triggered by proteins encoded by LEE region. We analyzed structurally and functionally the LEE region of aEPEC strains displaying LAL, AA, DA, and one nonadherent (NA) strain. The adherence characteristics and ability to cause A/E were investigated in epithelial cells. The displayed adherence patterns were independent of the cell line origin. A/E lesion was detected in some cellular lines after contact only with ALL- and AD-strains. LEE region presence was detected intact and real time PCR, microarray and immunodetection, in all samples tested. An EspFu-expressing plasmid was introduced in all strains, demonstrating no influence of this protein neither in the adherence patterns nor in the capacity to cause A/E of the LAL-, AA- and DA-strains. But, NA-strain expressed the LAL pattern and was able to cause A/E. Therefore, EspFu was shown to play a role in cell adhesion in addition to the establishment of the A/E lesion in vitro.
 
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Publishing Date
2010-10-21
 
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