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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2017.tde-09032017-134114
Document
Author
Full name
Thaís Akemi Amamura
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Isaac, Lourdes (President)
Carvalho, Enéas de
Loures, Flávio Vieira
Tambourgi, Denise Vilarinho
Title in Portuguese
Clivagem de proteínas do complexo de ataque à membrana do sistema complemento humano por proteases de leptospiras patogênicas.
Keywords in Portuguese
Evasão Imune
Leptospira
MAC
Proteases
Sistema Complemento
TCC
Abstract in Portuguese
A leptospirose é causada por bactérias que pertencem ao gênero Leptospira. Em um estudo realizado por nosso grupo, observou-se que as proteases secretadas por leptospiras patogênicas foram capazes de clivar a molécula C3 do Complemento e seus fragmentos C3b e iC3b, além de Fator B, C4b e C2. Neste trabalho expandimos a análise da atividade proteolítica sobre os componentes do Complexo de Ataque à Membrana (MAC): C6, C7, C8 e C9. Nós observamos que essas proteases clivam todos os componentes do MAC inclusive o complexo solúvel formado e que essas clivagens ocorrem de modo tempo-dependente. Além disso, as clivagens dessas moléculas ocorrem de modo seletivo, pois mesmo utilizando quantidades reduzidas de sobrenadantes ainda foi possível observar produtos de clivagem. A atividade proteolítica foi inibida pela 1,10fenantrolina, indicando a participação de metaloproteases. O reconhecimento de quais moléculas do MAC são clivadas por proteases de leptospiras patogênicas pode contribuir para o desenvolvimento de estratégias terapêuticas na infecção por estes patógenos.
Title in English
Cleavage of membrane attack complex proteins of human complement system by pathogenic leptospires proteases.
Keywords in English
Complement system
Immune evasion
Leptospira
MAC
Proteases
TCC
Abstract in English
Leptospirosis is a zoonosis caused by spirochetes from the genus Leptospira. In a previous study, our group observed that the proteases secreted by Pathogenic Leptospira were capable of cleaving C3 of Complement, as well as the fragments C3b and iC3b, Factor B (Alternative Pathway), C4 and C2 (Classical and Lectin Pathways). In this work, we analyze the activity of the leptospiral proteases on the components of Membrane Attack Complex (MAC). We observed that the protease cleaves all MAC components including soluble complex formed and that these cleavages occur in a time-dependent manner and in a selective way, since even when reduced quantities of supernatants were used, the cleavage products were still observed. The proteolytic activity was inhibited by 1,10phenanthroline, indicating the participation of metalloproteinases. The recognition that MAC molecules are cleaved by proteases of pathogenic leptospires can contribute to the development of therapeutic strategies for the infection by these pathogens.
 
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Publishing Date
2017-03-09
 
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