Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFPB |
Texto Completo: | https://repositorio.ufpb.br/jspui/handle/123456789/13138 |
Resumo: | Leishmaniasis is characterized as a complex of infectious-parasitic diseases caused by protozoa of the genus Leishmania. These diseases are present in 98 countries, where about 350 million people are at risk of infection. Visceral leishmaniasis (VL) is the most severe clinical form of the disease and induces immune responses mediated by distinct cell subpopulations, including invariant natural killer T cells (iNKT-CD3 + 6B11 +), regulatory T cells (Tregs - CD4 + CD25highFOXP3 +), which can be classified into natural Treg (CD4 + CD25highFOXP3 + CD45RA +) and memory Treg (CD4 + CD25highFOXP3 + CD45RA-). IL-10 is a key cytokine in the immunopathology of this disease and the mechanisms of action involved in the immunological modulation of human visceral leishmaniasis have not yet been well elucidated with respect to the subpopulations. Thus, our aim was to characterize iNKT and T regulatory cells (nTreg and mTreg) mechanisms during the diseases, and consequently evaluate the main cytokines produced. To achieve this goal, in the first part of this study, whole blood was collected from LV (n = 10) and uninfected (n = 7) patients, and the nTreg and mTreg subpopulations were evaluated with and without specific stimulation of the soluble antigen of Leishmania (SLA). The analysis of iNKT cells were also performed with and without specific stimulation of total Leishmania antigen (TLA) in whole blood samples in the LV (n = 12), nonendemic (CNE - n = 6) and in the endemic negative control group (CNegE - n = 6). All analyzes were performed by flow cytometry. Regarding the Tregs cells, the results demonstrated that: 1) A high number of memory Treg cells was observed when compared to the total number of natural Treg cells in all groups evaluated; 2) A significant reduction in CD39 / CD73 expression in mTreg cells was observed in the LV groups in the absence and / or presence of stimulus; 3) A decrease in CTLA-4 expression was observed in mTreg cells in the LV group in the absence of stimulation, when compared to the control group. However, SLA induced increased expression of CTLA-4 in patients with VL; 4) Significant changes in the production of IFN-? were not observed in any of the groups evaluated and 5) Low levels of IL-10 were observed in mTreg cells in the LV groups regardless of stimulus. In the iNKT cells, we observed that 6) the total ex vivo number of cells in the LV group did not change significantly in relation to the CNE and CNegE groups; 7) Regarding the recent activation profile of these cells, LV patients did not present significant changes in CD69 expression, as compared to the control groups both ex vivo and after culture in vitro (medium and TLA); 8) Following stimulation with L. infantum TLA, iNKT cells reduced IFN-? production. However, increased IFN-? production was observed in iNKT cells in the CNE group; 9) Regarding the production of IL-17, significant alterations were not observed, before or after TLA stimulation, in all the groups evaluated; 10) An increase in IL-10 production was observed in the CNE group when compared to CNegE and patients with VL independently stimulation. Thus, our results suggest that nTreg cells and iNKT cells, help maintain the effects of IL-10 on the chronicity of the disease. On the other hand, mTreg cells act counterbalancing the IL-10 mediated immunosuppression through mechanisms that aid in the induction of effector responses in the disease. However, despite the compensatory action of mTreg cells contributing to the establishment of VL control mechanisms, the pathogenic profile regulated by IL-10 still prevails in the disease. |
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Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humanaLeishmanioseInfecçãoLinfócito TLeishmaniasisInfectionT LymphocyteCNPQ::CIENCIAS BIOLOGICAS::FISIOLOGIALeishmaniasis is characterized as a complex of infectious-parasitic diseases caused by protozoa of the genus Leishmania. These diseases are present in 98 countries, where about 350 million people are at risk of infection. Visceral leishmaniasis (VL) is the most severe clinical form of the disease and induces immune responses mediated by distinct cell subpopulations, including invariant natural killer T cells (iNKT-CD3 + 6B11 +), regulatory T cells (Tregs - CD4 + CD25highFOXP3 +), which can be classified into natural Treg (CD4 + CD25highFOXP3 + CD45RA +) and memory Treg (CD4 + CD25highFOXP3 + CD45RA-). IL-10 is a key cytokine in the immunopathology of this disease and the mechanisms of action involved in the immunological modulation of human visceral leishmaniasis have not yet been well elucidated with respect to the subpopulations. Thus, our aim was to characterize iNKT and T regulatory cells (nTreg and mTreg) mechanisms during the diseases, and consequently evaluate the main cytokines produced. To achieve this goal, in the first part of this study, whole blood was collected from LV (n = 10) and uninfected (n = 7) patients, and the nTreg and mTreg subpopulations were evaluated with and without specific stimulation of the soluble antigen of Leishmania (SLA). The analysis of iNKT cells were also performed with and without specific stimulation of total Leishmania antigen (TLA) in whole blood samples in the LV (n = 12), nonendemic (CNE - n = 6) and in the endemic negative control group (CNegE - n = 6). All analyzes were performed by flow cytometry. Regarding the Tregs cells, the results demonstrated that: 1) A high number of memory Treg cells was observed when compared to the total number of natural Treg cells in all groups evaluated; 2) A significant reduction in CD39 / CD73 expression in mTreg cells was observed in the LV groups in the absence and / or presence of stimulus; 3) A decrease in CTLA-4 expression was observed in mTreg cells in the LV group in the absence of stimulation, when compared to the control group. However, SLA induced increased expression of CTLA-4 in patients with VL; 4) Significant changes in the production of IFN-? were not observed in any of the groups evaluated and 5) Low levels of IL-10 were observed in mTreg cells in the LV groups regardless of stimulus. In the iNKT cells, we observed that 6) the total ex vivo number of cells in the LV group did not change significantly in relation to the CNE and CNegE groups; 7) Regarding the recent activation profile of these cells, LV patients did not present significant changes in CD69 expression, as compared to the control groups both ex vivo and after culture in vitro (medium and TLA); 8) Following stimulation with L. infantum TLA, iNKT cells reduced IFN-? production. However, increased IFN-? production was observed in iNKT cells in the CNE group; 9) Regarding the production of IL-17, significant alterations were not observed, before or after TLA stimulation, in all the groups evaluated; 10) An increase in IL-10 production was observed in the CNE group when compared to CNegE and patients with VL independently stimulation. Thus, our results suggest that nTreg cells and iNKT cells, help maintain the effects of IL-10 on the chronicity of the disease. On the other hand, mTreg cells act counterbalancing the IL-10 mediated immunosuppression through mechanisms that aid in the induction of effector responses in the disease. However, despite the compensatory action of mTreg cells contributing to the establishment of VL control mechanisms, the pathogenic profile regulated by IL-10 still prevails in the disease.Fundação de Apoio à Pesquisa do Estado da Paraíba - FAPESQAs leishmanioses se caracterizam como um complexo de doenças infecto-parasitárias, causadas por protozoários do gênero Leishmania. Essas doenças estão presentes em 98 países, nos quais cerca de 350 milhões de pessoas encontram-se sob risco de infecção. A leishmaniose visceral (LV) é a forma clínica mais grave da doença e induz respostas imunes mediadas por distintas subpopulações celulares, incluindo células T natural killer invariantes (iNKT-CD3+6B11+), células T reguladoras (Tregs - CD4 + CD25highFOXP3 +), as quais podem ser classificadas em Treg natural (CD4 + CD25highFOXP3 + CD45RA +) e Treg de memória (CD4 + CD25highFOXP3 + CD45RA-). A IL-10 é uma citocina chave na imunopatologia dessa doença e os mecanismos de ação envolvidos na modulação imunológica da leishmaniose visceral humana, ainda não foram bem elucidados, no que diz respeito as subpopulações supracitadas. Assim, nosso objetivo foi caracterizar os mecanismos das células iNKT, células T reguladoras natural (nTreg) e de memória (mTreg) na doença e avaliar as principais citocinas produzidas por elas. Para atingir esse objetivo, na primeira parte desse trabalho coletou-se sangue total de pacientes LV (n=10) e indivíduos não infectados (n=7), e as subpopulações nTreg e mTreg foram avaliadas, com e sem estimulação específica do antígeno solúvel de Leishmania (SLA). A análise das células iNKT também foram realizadas, com e sem estimulação específica do antígeno total de Leishmania (TLA) em amostras de sangue total nos grupos de pacientes LV (n=12), em controles não endêmicos (CNE – n = 6) e no grupo controle negativo endêmico (CNegE – n = 6). Todas as análises foram realizadas através de citometria de fluxo. Em relação as células Tregs, os resultados demonstraram que: 1) Um número elevado de células Tregs de memória foi observado quando comparado ao número total de células Tregs naturais em todos os grupos avaliados; 2) Uma redução significativa na expressão de CD39/CD73 em células mTreg foi observada nos grupos LV na ausência e/ou presença de estímulo; 3) Uma diminuição na expressão de CTLA-4 foi observada em células mTreg no grupo LV na ausência de estímulo, quando comparada ao grupo controle. Entretanto, o SLA induziu aumento na expressão de CTLA-4, nos pacientes com LV; 4) Alterações significativas na produção de IFN-? não foram observadas em nenhum dos grupos avaliados e 5) Baixos níveis de IL-10 foram observados em células mTreg nos grupos LV independentemente de estímulo. Já nas células iNKT observamos que, 6) o número total ex vivo de células no grupo de pacientes LV não se alterou significativamente em relação aos grupos CNE e CNegE; 7) Em relação ao perfil de ativação recente dessas células, os pacientes LV não apresentaram alterações significativas na expressão de CD69, em comparação aos grupos controle tanto ex vivo como após a cultura in vitro (meio e TLA); 8) Após o estímulo com TLA de L. infantum as células iNKT reduziram a produção de IFN-?. No entanto, a maior produção de IFN-? foi observada em células iNKT no grupo CNE; 9) Em relação a produção de IL-17, alterações significativas não foram observadas, antes ou após a estimulação com o TLA, em todos os grupos avaliados; 10) Uma elevação na produção de IL-10, foi observada no grupo CNE quando comparado ao CNegE e pacientes com LV de forma independente a estimulação. Dessa forma, nossos resultados sugerem que as células nTreg e células iNKT, auxiliam na manutenção dos efeitos da IL-10 na cronicidade da doença. Por outro lado, as células mTreg atuam contrabalanceando a imunossupressão mediada pela IL-10, através de mecanismos que auxiliam na indução de respostas efetoras na doença. Entretanto, apesar da atuação compensatória das células mTreg contribuir para o estabelecimento de mecanismos de controle da LV, o perfil patogênico regulado pela IL-10 ainda prevalece na doença.Universidade Federal da ParaíbaBrasilCiências FisiológicasPrograma Multicêntrico de Pós-Graduação em Ciências FisiológicasUFPBLima, Tatjana Keesen de Souzahttp://lattes.cnpq.br/5504382837656473Peixoto, Rephany Fonseca2019-01-29T21:23:27Z2019-01-292019-01-29T21:23:27Z2018-04-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttps://repositorio.ufpb.br/jspui/handle/123456789/13138porAttribution-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nd/3.0/br/info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFPBinstname:Universidade Federal da Paraíba (UFPB)instacron:UFPB2019-01-29T21:23:27Zoai:repositorio.ufpb.br:123456789/13138Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufpb.br/PUBhttp://tede.biblioteca.ufpb.br:8080/oai/requestdiretoria@ufpb.br|| diretoria@ufpb.bropendoar:2019-01-29T21:23:27Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB)false |
dc.title.none.fl_str_mv |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
title |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
spellingShingle |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana Peixoto, Rephany Fonseca Leishmaniose Infecção Linfócito T Leishmaniasis Infection T Lymphocyte CNPQ::CIENCIAS BIOLOGICAS::FISIOLOGIA |
title_short |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
title_full |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
title_fullStr |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
title_full_unstemmed |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
title_sort |
Investigação da imunomodulação por subpopulações de linfócitos na Leishmaniose visceral humana |
author |
Peixoto, Rephany Fonseca |
author_facet |
Peixoto, Rephany Fonseca |
author_role |
author |
dc.contributor.none.fl_str_mv |
Lima, Tatjana Keesen de Souza http://lattes.cnpq.br/5504382837656473 |
dc.contributor.author.fl_str_mv |
Peixoto, Rephany Fonseca |
dc.subject.por.fl_str_mv |
Leishmaniose Infecção Linfócito T Leishmaniasis Infection T Lymphocyte CNPQ::CIENCIAS BIOLOGICAS::FISIOLOGIA |
topic |
Leishmaniose Infecção Linfócito T Leishmaniasis Infection T Lymphocyte CNPQ::CIENCIAS BIOLOGICAS::FISIOLOGIA |
description |
Leishmaniasis is characterized as a complex of infectious-parasitic diseases caused by protozoa of the genus Leishmania. These diseases are present in 98 countries, where about 350 million people are at risk of infection. Visceral leishmaniasis (VL) is the most severe clinical form of the disease and induces immune responses mediated by distinct cell subpopulations, including invariant natural killer T cells (iNKT-CD3 + 6B11 +), regulatory T cells (Tregs - CD4 + CD25highFOXP3 +), which can be classified into natural Treg (CD4 + CD25highFOXP3 + CD45RA +) and memory Treg (CD4 + CD25highFOXP3 + CD45RA-). IL-10 is a key cytokine in the immunopathology of this disease and the mechanisms of action involved in the immunological modulation of human visceral leishmaniasis have not yet been well elucidated with respect to the subpopulations. Thus, our aim was to characterize iNKT and T regulatory cells (nTreg and mTreg) mechanisms during the diseases, and consequently evaluate the main cytokines produced. To achieve this goal, in the first part of this study, whole blood was collected from LV (n = 10) and uninfected (n = 7) patients, and the nTreg and mTreg subpopulations were evaluated with and without specific stimulation of the soluble antigen of Leishmania (SLA). The analysis of iNKT cells were also performed with and without specific stimulation of total Leishmania antigen (TLA) in whole blood samples in the LV (n = 12), nonendemic (CNE - n = 6) and in the endemic negative control group (CNegE - n = 6). All analyzes were performed by flow cytometry. Regarding the Tregs cells, the results demonstrated that: 1) A high number of memory Treg cells was observed when compared to the total number of natural Treg cells in all groups evaluated; 2) A significant reduction in CD39 / CD73 expression in mTreg cells was observed in the LV groups in the absence and / or presence of stimulus; 3) A decrease in CTLA-4 expression was observed in mTreg cells in the LV group in the absence of stimulation, when compared to the control group. However, SLA induced increased expression of CTLA-4 in patients with VL; 4) Significant changes in the production of IFN-? were not observed in any of the groups evaluated and 5) Low levels of IL-10 were observed in mTreg cells in the LV groups regardless of stimulus. In the iNKT cells, we observed that 6) the total ex vivo number of cells in the LV group did not change significantly in relation to the CNE and CNegE groups; 7) Regarding the recent activation profile of these cells, LV patients did not present significant changes in CD69 expression, as compared to the control groups both ex vivo and after culture in vitro (medium and TLA); 8) Following stimulation with L. infantum TLA, iNKT cells reduced IFN-? production. However, increased IFN-? production was observed in iNKT cells in the CNE group; 9) Regarding the production of IL-17, significant alterations were not observed, before or after TLA stimulation, in all the groups evaluated; 10) An increase in IL-10 production was observed in the CNE group when compared to CNegE and patients with VL independently stimulation. Thus, our results suggest that nTreg cells and iNKT cells, help maintain the effects of IL-10 on the chronicity of the disease. On the other hand, mTreg cells act counterbalancing the IL-10 mediated immunosuppression through mechanisms that aid in the induction of effector responses in the disease. However, despite the compensatory action of mTreg cells contributing to the establishment of VL control mechanisms, the pathogenic profile regulated by IL-10 still prevails in the disease. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-04-25 2019-01-29T21:23:27Z 2019-01-29 2019-01-29T21:23:27Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufpb.br/jspui/handle/123456789/13138 |
url |
https://repositorio.ufpb.br/jspui/handle/123456789/13138 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Ciências Fisiológicas Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas UFPB |
publisher.none.fl_str_mv |
Universidade Federal da Paraíba Brasil Ciências Fisiológicas Programa Multicêntrico de Pós-Graduação em Ciências Fisiológicas UFPB |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da UFPB instname:Universidade Federal da Paraíba (UFPB) instacron:UFPB |
instname_str |
Universidade Federal da Paraíba (UFPB) |
instacron_str |
UFPB |
institution |
UFPB |
reponame_str |
Biblioteca Digital de Teses e Dissertações da UFPB |
collection |
Biblioteca Digital de Teses e Dissertações da UFPB |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da UFPB - Universidade Federal da Paraíba (UFPB) |
repository.mail.fl_str_mv |
diretoria@ufpb.br|| diretoria@ufpb.br |
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1801842942910922752 |