Apoptose na infecção experimental com Ehrlichia canis em cães domésticos

The Canine MonocyticEhrlichiosis is a zoonotic disease caused by Ehrlichiacanis, a Gram negative bacteria, with a worldwide distribution. Some dogs with ehrlichiosis become asymptomaticcarriers while others develop an acute disease with rapid death. Apoptosis can be important in the elimination of i...

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Autor: Irma Ximena Barbosa Sanchez
Tipo de documento: tese
Estado:Versão publicada
Data de publicação:2013
País:Brasil
Recursos:Universidade Federal de Minas Gerais (UFMG)
Repositório:Repositório Institucional da UFMG
Idioma:português
OAI Identifier:oai:repositorio.ufmg.br:1843/BUBD-9E3G95
Acesso em linha:http://hdl.handle.net/1843/BUBD-9E3G95
Access Level:Acceso aberto
Palavra-chave:Patologia
Doenças do cão
Apoptose
Ehrlichia canis
Inflamação
Cães
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oai_identifier_str oai:repositorio.ufmg.br:1843/BUBD-9E3G95
network_acronym_str BR
network_name_str Brasil
repository_id_str
dc.title.none.fl_str_mv Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
title Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
spellingShingle Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
Irma Ximena Barbosa Sanchez
Patologia
Doenças do cão
Patologia
Apoptose
Ehrlichia canis
Inflamação
Cães
title_short Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
title_full Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
title_fullStr Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
title_full_unstemmed Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
title_sort Apoptose na infecção experimental com Ehrlichia canis em cães domésticos
dc.creator.none.fl_str_mv Irma Ximena Barbosa Sanchez
author Irma Ximena Barbosa Sanchez
author_facet Irma Ximena Barbosa Sanchez
author_role author
dc.contributor.none.fl_str_mv Anilton Cesar Vasconcelos
Rosangela Z. Machado
Maria Norma Melo
Milene Alvarenga Rachid
Marlene Isabel Vargas Viloria
Joaquim Hernan Patarroyo Salcedo
Rosa Maria Esteves Arantes
dc.subject.por.fl_str_mv Patologia
Doenças do cão
Patologia
Apoptose
Ehrlichia canis
Inflamação
Cães
topic Patologia
Doenças do cão
Patologia
Apoptose
Ehrlichia canis
Inflamação
Cães
description The Canine MonocyticEhrlichiosis is a zoonotic disease caused by Ehrlichiacanis, a Gram negative bacteria, with a worldwide distribution. Some dogs with ehrlichiosis become asymptomaticcarriers while others develop an acute disease with rapid death. Apoptosis can be important in the elimination of intracellular pathogens in infections caused by Ehrlichia sp. and Anaplasma sp. occur modulation of apoptosis to prolong the survival of these organisms allowing their proliferation and spread, acting as a Trojan horse by inhibition the immune response. In this context apoptosis may contribute to the progression or regression of the lesions. To evaluate the role of apoptosis in Canine MonocyticEhrlichiosis, seven mongrel dogs, clinically healthy and free of ticks, aged among six and eight months, were randomly assigned into two groups: GI, Inoculated - with four dogs and GII, Control -with three dogs. Each dog was inoculated intravenously with 3 ml of either infected blood (strain Jaboticabal) in GI or sterile PBS in GII. Weekly and 35 days post-inoculation, blood samples were collected and subjected to nested reaction chain polymerase (nPCR) and immunofluorescence assay (IFA) for confirmation of infection. On day 36 post-inoculation, animals were euthanized and samples of spleen, lymph node, kidney and liver were collected and processed for TUNEL and immunohistochemical techniques (Bcl-2, Bax). Another set of the same samples, with bone marrow and heart were processed for gene expression of apoptotic molecules (Bax, Bcl-2, caspase-3, caspase 8 and caspase 9) by RTPCR. As verified by nPCR inoculated animals became positive for E. canis from the second week after inoculation. IFAT showed seropositive animals from the third week after inoculation. In the control group, all tests were negative for E. canis. Both pro and anti-apoptotic molecules showed higher expression of genes in tissues of inoculated animals than in the control group. TUNEL assay showed increased incidence of apoptosis in the group inoculated, with more intense labeling in spleen and lymph nodes, however the results of the immunohistochemistry for Bcl-2 and Bax indicated that the intrinsic pathway of apoptosis was not important. In conclusion we found that in the acute phase of the experimental disease,Ehrlichiacanis modulates apoptosis in different ways in different tissues and that apoptosis plays a role in disease progression.
publishDate 2013
dc.date.none.fl_str_mv 2013-06-07
2019-08-12T06:44:49Z
2019-08-12T06:44:49Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/BUBD-9E3G95
url http://hdl.handle.net/1843/BUBD-9E3G95
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
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)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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spelling Apoptose na infecção experimental com Ehrlichia canis em cães domésticosPatologiaDoenças do cãoPatologiaApoptoseEhrlichia canisInflamaçãoCãesThe Canine MonocyticEhrlichiosis is a zoonotic disease caused by Ehrlichiacanis, a Gram negative bacteria, with a worldwide distribution. Some dogs with ehrlichiosis become asymptomaticcarriers while others develop an acute disease with rapid death. Apoptosis can be important in the elimination of intracellular pathogens in infections caused by Ehrlichia sp. and Anaplasma sp. occur modulation of apoptosis to prolong the survival of these organisms allowing their proliferation and spread, acting as a Trojan horse by inhibition the immune response. In this context apoptosis may contribute to the progression or regression of the lesions. To evaluate the role of apoptosis in Canine MonocyticEhrlichiosis, seven mongrel dogs, clinically healthy and free of ticks, aged among six and eight months, were randomly assigned into two groups: GI, Inoculated - with four dogs and GII, Control -with three dogs. Each dog was inoculated intravenously with 3 ml of either infected blood (strain Jaboticabal) in GI or sterile PBS in GII. Weekly and 35 days post-inoculation, blood samples were collected and subjected to nested reaction chain polymerase (nPCR) and immunofluorescence assay (IFA) for confirmation of infection. On day 36 post-inoculation, animals were euthanized and samples of spleen, lymph node, kidney and liver were collected and processed for TUNEL and immunohistochemical techniques (Bcl-2, Bax). Another set of the same samples, with bone marrow and heart were processed for gene expression of apoptotic molecules (Bax, Bcl-2, caspase-3, caspase 8 and caspase 9) by RTPCR. As verified by nPCR inoculated animals became positive for E. canis from the second week after inoculation. IFAT showed seropositive animals from the third week after inoculation. In the control group, all tests were negative for E. canis. Both pro and anti-apoptotic molecules showed higher expression of genes in tissues of inoculated animals than in the control group. TUNEL assay showed increased incidence of apoptosis in the group inoculated, with more intense labeling in spleen and lymph nodes, however the results of the immunohistochemistry for Bcl-2 and Bax indicated that the intrinsic pathway of apoptosis was not important. In conclusion we found that in the acute phase of the experimental disease,Ehrlichiacanis modulates apoptosis in different ways in different tissues and that apoptosis plays a role in disease progression.A Erliquiose monocítica canina é uma zoonose causada pelaEhrlichia canis, bactéria Gram negativa de distribuição mundial. Alguns cães com erliquiose se tornam portadoresassintomáticos enquanto outros desenvolvem uma doençaaguda com morte rápida. A apoptose pode ser importante naeliminação de patógenos intracelulares, podendo, nas infecções porEhrlichiasp. eAnaplasmasp., ocorrer modulação daapoptose celular para prolongar a sobrevivência desses organismos, permitindo sua proliferação e disseminação, atuando como um cavalo de Tróia inibindo a resposta imune. Neste contexto, a apoptose pode contribuir indistintamente para a progressão ou regressão das lesões. Para avaliação do papel da apoptose na erliquiose monocítica canina, sete cães foram distribuídos em dois grupos. No Grupoinoculado, realizou-se a infecção por via intravenosa comsangue infectado com E. Canis (isolado Jaboticabal), sendorealizada a inoculação com PBS estéril nos animaispertencentes ao Grupo Controle. Semanalmente e até 35 diaspós-inoculação, amostras de sangue foram coletadas esubmetidas a n-PCR e reação de Imunofluorescência (RIFI) para confirmação da infecção. No 36° dia pós-inoculação, os animais foram eutanasiados, sendo amostras de baço, linfonodo, rim e fígado coletadas e processadas para as técnicas de TUNEL e imunohistoquímica (Bcl-2, Bax) e amostras dos mesmos, medula óssea e coração para expressão gênica de moléculas apoptóticas (Bax, Bcl-2, Caspase3, Caspase8 e Caspase 9) por RT-PCR. Verificou-se pelan-PCR que os animais inoculados se tornaram positivos paraE.canisa partir da segunda semana pós inoculação. Pela RIFI, verificou-seanimais com sorologia positiva a partir da terceira semanapós-inoculação. No grupo controle, todos os testes realizadosforam negativos para E. canis. Ambas as moléculas pro e antiapoptóticas avaliadas neste estudo mostraram mais alta expressão de genes em tecidos dos animais do grupo inoculado que nos animais do grupo controle. Apesar da reação de TUNELmostrar maior ocorrência de apoptose no Grupo Inoculado,sendo o baço e os linfonodos os órgãos que apresentarammaior marcação, os resultados da imunohistoquímica para Bcl-2 e Bax indicaram que a via intrínseca de apoptose não é tão importante nos órgãos analisados.Em conclusão nós encontramos que na fase aguda da doença experimental a Ehrlichiacanis modulaa apoptose por diferentes vias em tecidos distintos e que a apoptose tem um papel na progressão da doença.Universidade Federal de Minas GeraisUFMGAnilton Cesar VasconcelosRosangela Z. MachadoMaria Norma MeloMilene Alvarenga RachidMarlene Isabel Vargas ViloriaJoaquim Hernan Patarroyo SalcedoRosa Maria Esteves Arantes2019-08-12T06:44:49Z2019-08-12T06:44:49Z2013-06-07info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/1843/BUBD-9E3G95info:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGIrma Ximena Barbosa Sanchez2019-11-14T13:35:38Zoai:repositorio.ufmg.br:1843/BUBD-9E3G95Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2019-11-14T13:35:38Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
score 15,301603

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