Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi

Detalhes bibliográficos
Autor(a) principal: Thabata Lopes Alberto Duque
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/BUOS-97VFL9
Resumo: Chagas disease is caused by the intracellular protozoan parasite Trypanosoma cruzi. It remains a serious public health problem in Latin America, and the current treatment is not satisfactory. The disease morbidity is associated with cardiomyopathy, characterized by tissue damage, inflammatory infiltrates and fibrosis. Parasite internalization and its subsequent success in infection depend on the fusion of the parasitophorous vacuole (PV) to the lysosome, an important organelle related to endocytosis and autophagy. The autophagic pathway is the process involved in degradation of macromolecules and organelles. This pathway was recently associated with T. cruzi infection, but its effective participation as well as its functional role during the infection remain to be defined. Thus, this work aimed to evaluate the occurrence and role of autophagy of macrophages and heart muscle cells (HMC) during the T. cruzi infection. Pre-incubation with autophagic inducers (rapamycin and starvation medium DMEM -/-) reduced infection and endocytic index in both cells, while incubation after infection decreased the parasite replication only in HMC. Ultrastructural analyses of cells previously stimulated with rapamycin and DMEM -/-revealed typical autophagic features such as autophagosomes and concentric membranes. PV- autophagosomes association, suggestive of xenophagy, was also observed. Time- and cell-dependent increase in the autophagic protein LC3 was detected by immunofluorescence after autophagic induction. Immunonanogold electron microscopy showed an i ncrease in this protein expression after infection within autophagosomes. The cross-talk between autophagic pathway and lipid body biogenesis (important inflammatory organelles during infection) indicated a cell-dependent modulation. In conclusion, autophagy was characterized as an important process in response to the T. cruzi infection and seems to participate in the host resistance acting in the control of the infection in macrophages and cardiac cells.
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spelling Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruziAutofagiaMacrófagosCardiomiócitosTrypanosoma cruziTripanossoma cruziMacrófagosBiologia celularMiócitos cardíacosChagas disease is caused by the intracellular protozoan parasite Trypanosoma cruzi. It remains a serious public health problem in Latin America, and the current treatment is not satisfactory. The disease morbidity is associated with cardiomyopathy, characterized by tissue damage, inflammatory infiltrates and fibrosis. Parasite internalization and its subsequent success in infection depend on the fusion of the parasitophorous vacuole (PV) to the lysosome, an important organelle related to endocytosis and autophagy. The autophagic pathway is the process involved in degradation of macromolecules and organelles. This pathway was recently associated with T. cruzi infection, but its effective participation as well as its functional role during the infection remain to be defined. Thus, this work aimed to evaluate the occurrence and role of autophagy of macrophages and heart muscle cells (HMC) during the T. cruzi infection. Pre-incubation with autophagic inducers (rapamycin and starvation medium DMEM -/-) reduced infection and endocytic index in both cells, while incubation after infection decreased the parasite replication only in HMC. Ultrastructural analyses of cells previously stimulated with rapamycin and DMEM -/-revealed typical autophagic features such as autophagosomes and concentric membranes. PV- autophagosomes association, suggestive of xenophagy, was also observed. Time- and cell-dependent increase in the autophagic protein LC3 was detected by immunofluorescence after autophagic induction. Immunonanogold electron microscopy showed an i ncrease in this protein expression after infection within autophagosomes. The cross-talk between autophagic pathway and lipid body biogenesis (important inflammatory organelles during infection) indicated a cell-dependent modulation. In conclusion, autophagy was characterized as an important process in response to the T. cruzi infection and seems to participate in the host resistance acting in the control of the infection in macrophages and cardiac cells.A doença de Chagas é causada pelo protozoário Trypanosoma cruzi e representa um sério problema de saúde na América Latina, sendo considerada uma doença negligenciada cujo tratamento atual é insatisfatório e limitado. A sua principal causa de morbidade é a cardiomiopatia, caracterizada por danos nas células cardíacas e presença de infiltrados inflamatórios e fibrose. A interação parasito-célula hospedeira e o consequente estabelecimento da infecção dependem da fusão do vacúolo parasitóforo (VP) com o lisossomo, organela relacionada à endocitose e autofagia. A via autofágica consiste em um importante processo de degradação de macromoléculas e organelas, tendo sido recentemente associada à infecção pelo T. cruzi. No entanto, a participação efetiva da autofagia, assim como seu papel na infecção, não são conhecidas. Nesse contexto, o presente trabalho teve como objetivo analisar a participação da autofagia durante a infecção de macrófagos e células cardíacas (HMC) por T. cruzi. A pré-incubação com indutor farmacológico de autofagia (rapamicina) ou com meio de estresse nutricional (DMEM-/-) reduziu a infecção e a taxa endocítica em ambas as células, enquanto que a incubação posterior à infecção foi capaz de diminuir a proliferação do parasito apenas em HMC. A análise ultraestrutural apontou características autofágicas típicas como a predominância de autofagossomos e perfis de membrana concêntrica em células estimuladas pré-infecção, além da associação do VP ao autofagossomo, sugestivo de xenofagia. A imunomarcação para a proteína de autofagia LC3 foi mais abundante em células incubadas com os indutores de autofagia, variando a associação ao parasito nos diferentes tipos celulares e tempos. A imunolocalização por microscopia eletrônica revelou um aumento na expressão desta proteína após a infecção, localizadas no interior de autofagossomos. A análise da interrelação da via autofágica com a biogênese de corpúsculos lipídicos (importantes mediadores inflamatórios durante a infecção) indicou inibição da formação destas organelas em macrófagos, enquanto em HMC não ocorreu alteração do número. Os dados do presente trabalho identificam a autofagia como um processo importante que ocorre em resposta à infecção por T. cruzi e que parece atuar na resistência do hospedeiro, controlando a infecção em macrófagos e células cardíacas.Universidade Federal de Minas GeraisUFMGRossana Correa Netto de MeloRubem Figueiredo Sadok Menna BarretoAnnamaria Ravara VagoElizabeth Ribeiro da SilvaRubem Figueiredo Sadok Menna BarretoThabata Lopes Alberto Duque2019-08-12T00:31:38Z2019-08-12T00:31:38Z2013-02-28info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/1843/BUOS-97VFL9info:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2019-11-14T10:04:58Zoai:repositorio.ufmg.br:1843/BUOS-97VFL9Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2019-11-14T10:04:58Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
title Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
spellingShingle Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
Thabata Lopes Alberto Duque
Autofagia
Macrófagos
Cardiomiócitos
Trypanosoma cruzi
Tripanossoma cruzi
Macrófagos
Biologia celular
Miócitos cardíacos
title_short Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
title_full Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
title_fullStr Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
title_full_unstemmed Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
title_sort Avaliação da participação da autofagia de células hospedeiras durante a infecção experimental por Trypanosoma cruzi
author Thabata Lopes Alberto Duque
author_facet Thabata Lopes Alberto Duque
author_role author
dc.contributor.none.fl_str_mv Rossana Correa Netto de Melo
Rubem Figueiredo Sadok Menna Barreto
Annamaria Ravara Vago
Elizabeth Ribeiro da Silva
Rubem Figueiredo Sadok Menna Barreto
dc.contributor.author.fl_str_mv Thabata Lopes Alberto Duque
dc.subject.por.fl_str_mv Autofagia
Macrófagos
Cardiomiócitos
Trypanosoma cruzi
Tripanossoma cruzi
Macrófagos
Biologia celular
Miócitos cardíacos
topic Autofagia
Macrófagos
Cardiomiócitos
Trypanosoma cruzi
Tripanossoma cruzi
Macrófagos
Biologia celular
Miócitos cardíacos
description Chagas disease is caused by the intracellular protozoan parasite Trypanosoma cruzi. It remains a serious public health problem in Latin America, and the current treatment is not satisfactory. The disease morbidity is associated with cardiomyopathy, characterized by tissue damage, inflammatory infiltrates and fibrosis. Parasite internalization and its subsequent success in infection depend on the fusion of the parasitophorous vacuole (PV) to the lysosome, an important organelle related to endocytosis and autophagy. The autophagic pathway is the process involved in degradation of macromolecules and organelles. This pathway was recently associated with T. cruzi infection, but its effective participation as well as its functional role during the infection remain to be defined. Thus, this work aimed to evaluate the occurrence and role of autophagy of macrophages and heart muscle cells (HMC) during the T. cruzi infection. Pre-incubation with autophagic inducers (rapamycin and starvation medium DMEM -/-) reduced infection and endocytic index in both cells, while incubation after infection decreased the parasite replication only in HMC. Ultrastructural analyses of cells previously stimulated with rapamycin and DMEM -/-revealed typical autophagic features such as autophagosomes and concentric membranes. PV- autophagosomes association, suggestive of xenophagy, was also observed. Time- and cell-dependent increase in the autophagic protein LC3 was detected by immunofluorescence after autophagic induction. Immunonanogold electron microscopy showed an i ncrease in this protein expression after infection within autophagosomes. The cross-talk between autophagic pathway and lipid body biogenesis (important inflammatory organelles during infection) indicated a cell-dependent modulation. In conclusion, autophagy was characterized as an important process in response to the T. cruzi infection and seems to participate in the host resistance acting in the control of the infection in macrophages and cardiac cells.
publishDate 2013
dc.date.none.fl_str_mv 2013-02-28
2019-08-12T00:31:38Z
2019-08-12T00:31:38Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
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dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/BUOS-97VFL9
url http://hdl.handle.net/1843/BUOS-97VFL9
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language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
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institution UFMG
reponame_str Repositório Institucional da UFMG
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