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Master's Dissertation
DOI
https://doi.org/10.11606/D.25.2009.tde-03072009-103411
Document
Author
Full name
Hayana Ramos Lima
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 2009
Supervisor
Committee
Campanelli, Ana Paula (President)
Garlet, Ana Paula Favaro Trombone
Santos, Carlos Ferreira dos
Title in Portuguese
Caracterização do papel do receptor do tipo Toll 4 (TLR4) em infecção por Aggregatibacter actinomycetemcomitans
Keywords in Portuguese
Aggregatibacter actinomycetemcomitans
Doença periodontal
Receptor do tipo Toll 4
Abstract in Portuguese
Os tecidos periodontais estão em confronto continuo com microorganismos capazes de disparar mecanismos da resposta imune inata, dando origem ao infiltrado inflamatório. Estudos recentes mostraram a importancia dos receptores do tipo Toll (TLRs) na fase inicial de reconhecimento de diferentes patogenos. A participação de receptores tipo Toll (TLRs) na resposta de neutrófilos e macrófagos frente a periodontopatógenos precisa ser determinada. Nesse estudo procuramos caracterizar o infiltrado inflamatório presente no peritônio de animais deficientes de TLR4-/-, avaliar a atividade fagocítica, bem como a produção de óxido nítrico (NO) e a atividade de mieloperoxidase (MPO) no curso da infecção por Aggregatibacter actinomycetemcomitans. A ausência de TLR4 não influenciou a quimiotaxia de neutrófilos e macrófagos para o local da infecção, a produção de óxido nítrico, a atividade de MPO e a viabilidade celular. No entanto, neutrófilos e macrófagos de animais TLR4-/- apresentaram menor atividade fagocítica quando comparado ao grupo controle (camundongos WT). Em relação a doença periodontal induzida experimentalmente com Aggregatibacter actinomycetemcomitans em camundongos deficientes de TLR4, os resultados mostraram que 100% dos animais deficientes de TLR4 sobreviveram a infecção durante o período de observação. Em relação a análise de perda óssea, os dados revelaram uma menor perda progressiva de osso alveolar na região dos molares de animais deficientes de TLR4. A ausência do receptor interferiu na disseminação da bactéria, uma vez que se observou um grande número de bacilos no linfonodo e baco dos animais que não expressaram TLR4, diferente do observado para os animais selvagens (WT). Os resultados indicam a importância da sinalização via TLR4 durante a resposta imune contra Aggregatibacter actinomycetemcomitans.
Title in English
The role of TLR4 (Toll like receptor 4) in the recognition of Aggregatibacter actinomycetemcomitans
Keywords in English
Aggregatibacter actinomycetemcomitans
Periodontal disease
Toll like receptor 4 (TLR4)
Abstract in English
Aggregatibacter actinomycetemcomitans is an oral gram negative bacteria that contributes to periodontitis progression. Isolated antigens from A. actinomycetemcomitans could be activating innate immune cells through Toll-like receptors (TLRs), molecules that recognize structural components conserved among microorganisms. In this study, we evaluated the role of TLR4 in the recognition of Aggregatibacter actinomycetemcomitans. Neutrophils and macrophage from TLR4 deficient mice and WT mice were collected and used for the subsequent assays. The phagocytosis of leukocytes against A. actinomycetemcomitans and the presence of apoptotic cells were determined by flow cytometry. The in vivo and in vitro production of NO and MPO was evaluated 24h after A. actinomycetemcomitans challenge. In addition, we examined the mechanisms that modulate the outcome of A. actinomycetemcomitans-induced periodontal disease in TLR4-/- mice. The results show that inflammatory cells influx in peritoneal cavity of TLR4-/- mice was similar to that observed into their littermate controls. The phagocytic activity was diminished by cells from TLR4-/- mice. In addition, we did not observe difference in NO and MPO production and the frequency of apoptotic cells between cells from TLR4-/- and WT mice. The results showed that TLR4-deficient mice developed less severe periodontitis after A. actinomycetemcomitans infection, characterized by significantly lower bone loss and inflammatory cell migration to periodontal tissues. Together, these data demonstrate the role TLR4 signals for neutrophils activation after A. actinomycetemcomitans infection and development of periodontal disease.
 
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HayanaRamosLima.pdf (2.29 Mbytes)
Publishing Date
2009-07-07
 
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