Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase
Autor(a) principal: | |
---|---|
Data de Publicação: | 2006 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UERJ |
Texto Completo: | http://www.bdtd.uerj.br/handle/1/8808 |
Resumo: | Leprosy presents an ample spectrum of clinical forms with also histopathological differences. In this context, skin macrophages from paucibacillary and multibacillary patients have a different response to Mycobacterium leprae infection. In addition, the relationship of macrophage with others skin cells may have a key role in the development of local immunological response in these groups of patients. This way, the functional and morphological characterization of these macrophages is crucial for a better understanding of tissue lesion in leprosy. By enzymatic dissociation, cells from paucibacillary (n=3), multibacillary patients with (ENL; n=5) and without reaction (n=13) were isolated. For the morphologic and functional analysis of this sample, Giemsa and Ziehl-Neelsen staining was used, as well as, flow cytometry, immunocytochemistry, ELISA and real time PCR. In parallel, skin fragments used for culture, were stored and processed for histopathological and real time RT-PCR analysis. Approximately 40-50% of the cell population of the groups was CD14+, characteristic of macrophages. The CD3+ cells (lymphocyte) analysis showed a major percentage of this cell population in paucibacillary patients. The inverse was observed for CD56+ cells (NK cells), where a greater percentage was verified in multibacillary patients. Moreover, the analysis of TLR-2 and TLR-4 expression demonstrated that the percentage of TLR-2+ was higher than TLR-4+ in paucibacillary and multibacillary patients. In patients with ENL a higher percentage of TLR4+ cells was observed which could be related with the hiper-reactivity of the immune inflammatory response. Giemsa staining showed morphological characteristics of macrophages during the whole period of culture and Ziehl-Neelsen s the bacilli detection respectively in these paucibacillary patients cells. Moreover, the cells presented intense marking for CD68 in immunocytochemistry. During the period of culture the cells secreted TNFα, IFNγ and IL-10, as well as inflammatory mediators. The gene expression for these same citokines was evaluated by real time PCR, and showed an increase in the TNFα and IFNγ expression in cells of paucibacillary compared with the multibacillary patients and a higher expression of IL-10 in multibacillary. A similar profile to the one of the multibacillary patients was observed in the patients with ENL and in the skin fragments. The analysis of the TNFα secretion, IFNγ and IL-10 was carried out and the same profile of the analysis for RT-PCR was observed. These data confirm isolated different profiles of Mфfrom paucibacillary and multibacillary patient s skin in the same way that it is observed in the tecidual injury in leprosy. |
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Sampaio, Elizabeth Pereirahttp://lattes.cnpq.br/8058296962302730Sarno, Euzenir Nuneshttp://lattes.cnpq.br/3330957069068672Cruz, Alda Maria dahttp://lattes.cnpq.br/2028714603825846Antunes, Sergio Luiz Gomeshttp://lattes.cnpq.br/6330876489112189Costa, Andréa Monte Altohttp://lattes.cnpq.br/2742511199834466http://lattes.cnpq.br/1757761916657648Moura, Danielle Fonseca de2021-01-05T19:43:06Z2018-05-022006-10-09MOURA, Danielle Fonseca de. Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase. 2006. 96 f. Dissertação (Mestrado em Ciências Médicas) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2006.http://www.bdtd.uerj.br/handle/1/8808Leprosy presents an ample spectrum of clinical forms with also histopathological differences. In this context, skin macrophages from paucibacillary and multibacillary patients have a different response to Mycobacterium leprae infection. In addition, the relationship of macrophage with others skin cells may have a key role in the development of local immunological response in these groups of patients. This way, the functional and morphological characterization of these macrophages is crucial for a better understanding of tissue lesion in leprosy. By enzymatic dissociation, cells from paucibacillary (n=3), multibacillary patients with (ENL; n=5) and without reaction (n=13) were isolated. For the morphologic and functional analysis of this sample, Giemsa and Ziehl-Neelsen staining was used, as well as, flow cytometry, immunocytochemistry, ELISA and real time PCR. In parallel, skin fragments used for culture, were stored and processed for histopathological and real time RT-PCR analysis. Approximately 40-50% of the cell population of the groups was CD14+, characteristic of macrophages. The CD3+ cells (lymphocyte) analysis showed a major percentage of this cell population in paucibacillary patients. The inverse was observed for CD56+ cells (NK cells), where a greater percentage was verified in multibacillary patients. Moreover, the analysis of TLR-2 and TLR-4 expression demonstrated that the percentage of TLR-2+ was higher than TLR-4+ in paucibacillary and multibacillary patients. In patients with ENL a higher percentage of TLR4+ cells was observed which could be related with the hiper-reactivity of the immune inflammatory response. Giemsa staining showed morphological characteristics of macrophages during the whole period of culture and Ziehl-Neelsen s the bacilli detection respectively in these paucibacillary patients cells. Moreover, the cells presented intense marking for CD68 in immunocytochemistry. During the period of culture the cells secreted TNFα, IFNγ and IL-10, as well as inflammatory mediators. The gene expression for these same citokines was evaluated by real time PCR, and showed an increase in the TNFα and IFNγ expression in cells of paucibacillary compared with the multibacillary patients and a higher expression of IL-10 in multibacillary. A similar profile to the one of the multibacillary patients was observed in the patients with ENL and in the skin fragments. The analysis of the TNFα secretion, IFNγ and IL-10 was carried out and the same profile of the analysis for RT-PCR was observed. These data confirm isolated different profiles of Mфfrom paucibacillary and multibacillary patient s skin in the same way that it is observed in the tecidual injury in leprosy.A hanseníase apresenta um amplo espectro de formas clínicas que possuem diferenças histopatológicas entre si. Neste contexto, os macrófagos (Mф) da pele de pacientes paucibacilares e multibacilares possuem uma resposta diferenciada à infecção pelo Mycobacterium leprae. Além disso, a relação do Mф com outras células da pele pode exercer papel chave no desenvolvimento da resposta imunológica local nestes grupos de pacientes. Desta forma, a caracterização morfológica e funcional destes Mф é crucial para um melhor entendimento da lesão tecidual na hanseníase. Por dissociação enzimática, células da lesão de pacientes paucibacilares (n=3), multibacilares (n=13) e durante o episódio reacional (ENL; n=5) foram isoladas. Para a análise morfológica e funcional desta amostra foram realizadas colorações por Giemsa e Ziehl-Neelsen, assim como, citometria de fluxo, imunocitoquímica, ELISA e PCR em tempo real. Em paralelo, fragmentos de pele utilizados para a cultura, foram armazenados e processados para análise histopatológica e avaliação por RT-PCR em tempo real. Aproximadamente 40-50% da população nos grupos utilizados era CD14+, característico de Mф. A análise das células e CD3+ (linfócito) mostraram uma maior porcentagem desta população em pacientes paucibacilares. O inverso foi observado para células CD56+ (células NK) onde a maior porcentagem foi verificada em multibacilares. Além disso, também foi analisada a expressão de TLR-2 e TLR-4, onde foi verificado que a porcentagem de células TLR2+ era maior do que a TLR-4+ em pacientes pauci e multibacilares. No entanto, em pacientes com ENL foi encontrada uma maior porcentagem de células TLR4+ o que pode estar relacionado com a hiper reatividade da resposta imuno-inflamatória. Coloração por Giemsa e Ziehl-Neelsen mostraram características morfológicas de M durante o tempo de cultura e a detecção de bacilos nestas células em pacientes multibacilares respectivamente. Além disso, as células apresentavam intensa marcação para CD68 por imunocitoquímica. Durante o período de cultura as células secretaram TNFα, IFNγ e IL-10, assim como mediadores inflamatórios. A expressão gênica para as mesmas citocinas analisadas foi feita por PCR em tempo real, onde ocorreu um aumento na expressão de TNFα IFNγ em células de paucibacilares comparados a multibacilares e uma maior expressão de IL-10 em multibacilares. Um perfil semelhante ao dos pacientes multibacilares foi observado nos pacientes com ENL e em biópsias de pele. A análise da secreção de TNFα, IFNγ e IL-10 foi realizada e o mesmo perfil da análise por RT-PCR foi mantido. Estes dados confirmam perfis diferentes de Mф isolados da pele de pacientes paucibacilares e multibacilares da mesma forma que é observado na lesão tecidual na hanseníase.Submitted by Boris Flegr (boris@uerj.br) on 2021-01-05T19:43:06Z No. of bitstreams: 1 DISSERTACAO_FINAL_PUBLICADA_Danielle_ Fonseca_de_Moura.pdf: 3384501 bytes, checksum: 394af32770de4eef82f715122eb091a6 (MD5)Made available in DSpace on 2021-01-05T19:43:06Z (GMT). No. of bitstreams: 1 DISSERTACAO_FINAL_PUBLICADA_Danielle_ Fonseca_de_Moura.pdf: 3384501 bytes, checksum: 394af32770de4eef82f715122eb091a6 (MD5) Previous issue date: 2006-10-09application/pdfporUniversidade do Estado do Rio de JaneiroPrograma de Pós-Graduação em Ciências MédicasUERJBRCentro Biomédico::Faculdade de Ciências MédicasLeprosyMacrophagesSkinHanseníaseMacrófagosPeleHanseníaseMacrófagosPele DoençasMycobacterium lepraeCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIACaracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníaseMorphological and functional characterization of inflammatory infiltrated cells on skin leprosy lesionsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UERJinstname:Universidade do Estado do Rio de Janeiro (UERJ)instacron:UERJORIGINALDISSERTACAO_FINAL_PUBLICADA_Danielle_ Fonseca_de_Moura.pdfapplication/pdf3384501http://www.bdtd.uerj.br/bitstream/1/8808/1/DISSERTACAO_FINAL_PUBLICADA_Danielle_+Fonseca_de_Moura.pdf394af32770de4eef82f715122eb091a6MD511/88082024-02-26 16:00:06.17oai:www.bdtd.uerj.br:1/8808Biblioteca Digital de Teses e Dissertaçõeshttp://www.bdtd.uerj.br/PUBhttps://www.bdtd.uerj.br:8443/oai/requestbdtd.suporte@uerj.bropendoar:29032024-02-26T19:00:06Biblioteca Digital de Teses e Dissertações da UERJ - Universidade do Estado do Rio de Janeiro (UERJ)false |
dc.title.por.fl_str_mv |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
dc.title.alternative.eng.fl_str_mv |
Morphological and functional characterization of inflammatory infiltrated cells on skin leprosy lesions |
title |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
spellingShingle |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase Moura, Danielle Fonseca de Leprosy Macrophages Skin Hanseníase Macrófagos Pele Hanseníase Macrófagos Pele Doenças Mycobacterium leprae CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
title_short |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
title_full |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
title_fullStr |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
title_full_unstemmed |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
title_sort |
Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase |
author |
Moura, Danielle Fonseca de |
author_facet |
Moura, Danielle Fonseca de |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Sampaio, Elizabeth Pereira |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8058296962302730 |
dc.contributor.advisor-co1.fl_str_mv |
Sarno, Euzenir Nunes |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/3330957069068672 |
dc.contributor.referee1.fl_str_mv |
Cruz, Alda Maria da |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/2028714603825846 |
dc.contributor.referee2.fl_str_mv |
Antunes, Sergio Luiz Gomes |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/6330876489112189 |
dc.contributor.referee3.fl_str_mv |
Costa, Andréa Monte Alto |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/2742511199834466 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/1757761916657648 |
dc.contributor.author.fl_str_mv |
Moura, Danielle Fonseca de |
contributor_str_mv |
Sampaio, Elizabeth Pereira Sarno, Euzenir Nunes Cruz, Alda Maria da Antunes, Sergio Luiz Gomes Costa, Andréa Monte Alto |
dc.subject.eng.fl_str_mv |
Leprosy Macrophages Skin |
topic |
Leprosy Macrophages Skin Hanseníase Macrófagos Pele Hanseníase Macrófagos Pele Doenças Mycobacterium leprae CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
dc.subject.por.fl_str_mv |
Hanseníase Macrófagos Pele Hanseníase Macrófagos Pele Doenças Mycobacterium leprae |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
description |
Leprosy presents an ample spectrum of clinical forms with also histopathological differences. In this context, skin macrophages from paucibacillary and multibacillary patients have a different response to Mycobacterium leprae infection. In addition, the relationship of macrophage with others skin cells may have a key role in the development of local immunological response in these groups of patients. This way, the functional and morphological characterization of these macrophages is crucial for a better understanding of tissue lesion in leprosy. By enzymatic dissociation, cells from paucibacillary (n=3), multibacillary patients with (ENL; n=5) and without reaction (n=13) were isolated. For the morphologic and functional analysis of this sample, Giemsa and Ziehl-Neelsen staining was used, as well as, flow cytometry, immunocytochemistry, ELISA and real time PCR. In parallel, skin fragments used for culture, were stored and processed for histopathological and real time RT-PCR analysis. Approximately 40-50% of the cell population of the groups was CD14+, characteristic of macrophages. The CD3+ cells (lymphocyte) analysis showed a major percentage of this cell population in paucibacillary patients. The inverse was observed for CD56+ cells (NK cells), where a greater percentage was verified in multibacillary patients. Moreover, the analysis of TLR-2 and TLR-4 expression demonstrated that the percentage of TLR-2+ was higher than TLR-4+ in paucibacillary and multibacillary patients. In patients with ENL a higher percentage of TLR4+ cells was observed which could be related with the hiper-reactivity of the immune inflammatory response. Giemsa staining showed morphological characteristics of macrophages during the whole period of culture and Ziehl-Neelsen s the bacilli detection respectively in these paucibacillary patients cells. Moreover, the cells presented intense marking for CD68 in immunocytochemistry. During the period of culture the cells secreted TNFα, IFNγ and IL-10, as well as inflammatory mediators. The gene expression for these same citokines was evaluated by real time PCR, and showed an increase in the TNFα and IFNγ expression in cells of paucibacillary compared with the multibacillary patients and a higher expression of IL-10 in multibacillary. A similar profile to the one of the multibacillary patients was observed in the patients with ENL and in the skin fragments. The analysis of the TNFα secretion, IFNγ and IL-10 was carried out and the same profile of the analysis for RT-PCR was observed. These data confirm isolated different profiles of Mфfrom paucibacillary and multibacillary patient s skin in the same way that it is observed in the tecidual injury in leprosy. |
publishDate |
2006 |
dc.date.issued.fl_str_mv |
2006-10-09 |
dc.date.available.fl_str_mv |
2018-05-02 |
dc.date.accessioned.fl_str_mv |
2021-01-05T19:43:06Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
MOURA, Danielle Fonseca de. Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase. 2006. 96 f. Dissertação (Mestrado em Ciências Médicas) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2006. |
dc.identifier.uri.fl_str_mv |
http://www.bdtd.uerj.br/handle/1/8808 |
identifier_str_mv |
MOURA, Danielle Fonseca de. Caracterização morfológica e funcional de células do infiltrado inflamatório na lesão da hanseníase. 2006. 96 f. Dissertação (Mestrado em Ciências Médicas) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2006. |
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http://www.bdtd.uerj.br/handle/1/8808 |
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Universidade do Estado do Rio de Janeiro |
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UERJ |
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BR |
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Centro Biomédico::Faculdade de Ciências Médicas |
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Universidade do Estado do Rio de Janeiro |
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Biblioteca Digital de Teses e Dissertações da UERJ |
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