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Doctoral Thesis
DOI
10.11606/T.41.2008.tde-18072008-155232
Document
Author
Full name
Enéas de Carvalho
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Ho, Paulo Lee (President)
Brito, Thales de
Silva, Aline Maria da
Soares Netto, Luis Eduardo
Yasuda, Paulo Hideki
Title in Portuguese
Análise das proteínas de Leptospira com possível papel hemolítico através de expressão recombinante: detecção de expressão nativa, atividade biológica e potencial vacinal
Keywords in Portuguese
Leptospira
Adesão
Hemolisina
Leptospirose
Vacina
Abstract in Portuguese
A leptospirose é considerada a zoonose mais difundida do mundo, assim como uma doença reemergente. Esta enfermidade, causada por bactérias patogênicas do gênero Leptospira, possui altas taxas de infecção em países em desenvolvimento, ocasionando graves prejuízos econômicos e de saúde pública. Até o momento, não existem vacinas humanas licenciadas contra leptospirose. Após o seqüenciamento do genoma de três espécies de Leptospiras vários genes foram apontados como candidatos vacinais promissores. Uma categoria importante de genes candidatos são aqueles com possível atividade hemolítica. Neste trabalho, clonamos e expressamos diversas proteínas com possível atuação hemolítica. As proteínas recombinantes obtidas, no entanto, não exibiram atividade hemolítica. Uma destas proteínas, TlyC, foi investigada quanto à sua capacidade de interagir com os componentes da matriz extracelular (MEC). Os resultados obtidos indicam que TlyC liga-se com alta afinidade a diversos componentes da MEC, e que esta proteína é capaz de inibir competitivamente a adesão de Leptospiras à um material biológico que se assemelha à MEC. A transcrição e expressão destas proteínas foi detectada em cultura de Leptospira. Algumas das proteínas recombinantes foram utilizadas em um desafio animal contra leptospirose, mas nenhum delas foi protetora. Concluímos que estas proteínas não parecem ser bons candidatos vacinais e que TlyC é uma proteína que interage com componentes da MEC.
Title in English
Analysis of the Leptospira proteins with putative hemolytic role thorough recombinant expression: detection of native expression, biological activity and vaccine potential
Keywords in English
Leptospira
Adhesion
Hemolysin
Leptospirosis
Vaccine
Abstract in English
Leptospirosis is considered the most disseminated zoonosis of the world, and also a reemerging disease. This disease, caused by pathogenic bacteria of the genus Leptospira, has high rates of infection in developing countries, leading to severe economic and medical costs. There is not a licensed vaccine against leptospirosis for human use. After the genome sequencing of three species of leptospires, several genes were pointed to be promising vaccinal candidates. An important category of these candidates are those with putative hemolytic activity. In this work, we cloned and expressed some proteins with putative hemolytic activity. The recombinant proteins obtained, however, did not show hemolytic activity. One of these proteins, TlyC, was investigated with regard to its possible ability to interact to extracellular matrix (ECM) components. The results obtained indicate that TlyC binds with high affinity to several ECM components and that this protein can inhibit the Leptospira bind to a biological material that ressambles the ECM. The transcription and expression of these proteins were detected in leptospires cultures. Some of the recombinant proteins were used in an animal challenge against leptospirosis, but none of them were protective. We conclude that these proteins do not seem to be good vaccine candidates and that TlyC is a protein that interacts with the ECM and its components.
 
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Publishing Date
2008-07-25
 
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