Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFMG |
Texto Completo: | https://doi.org/10.1016/j.exppara.2020.108060 http://hdl.handle.net/1843/54632 |
Resumo: | Amoebic keratitis (AK) is a sight-threatening infection characterized by a severe inflammation of the cornea, caused by the free-living protozoan of the genus Acanthamoeba. Identification of amoebic proteins involved in AK pathogenesis may help to elucidate molecular mechanisms of infection and contribute to indicate diagnostic and therapeutic targets. In this study, we evaluated changes in the expression profile of Acanthamoeba proteins triggered by the invasive process, using an approach involving two-dimensional polyacrylamide gel electrophoresis (2DE PAGE), followed by mass spectrometry identification (ESI-IT-TOF LC-MSn). AK was induced by intrastromal inoculation in Wistar rats, using trophozoites from a T4 genotype, human case-derived A. castellanii strain under prolonged axenic culture. Cultures re-isolated from the lesions after two successive passages in the animals were used as biological triplicate for proteomic experiments. Analysis of the protein profile comparing long-term and re-isolated cultures indicated 62 significant spots, from which 27 proteins could be identified in the Acanthamoeba proteome database. Five of them (Serpin, Carboxypeptidase A1, Hypothetical protein, Calponin domain-containing protein, aldo/keto reductase) were exclusively found in the re-isolated trophozoites. Our analysis also revealed that a concerted modulation of several biochemical pathways is triggered when A. castellanii switches from a free-living style to a parasitic mode, including energetic metabolism, proteolytic activity, control of gene expression, protein degradation and methylation of DNA, which may be also involved in gain of virulence in an animal model of AK. |
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2023-06-07T11:29:10Z2023-06-07T11:29:10Z2021-0222119https://doi.org/10.1016/j.exppara.2020.1080600753-3322http://hdl.handle.net/1843/54632Amoebic keratitis (AK) is a sight-threatening infection characterized by a severe inflammation of the cornea, caused by the free-living protozoan of the genus Acanthamoeba. Identification of amoebic proteins involved in AK pathogenesis may help to elucidate molecular mechanisms of infection and contribute to indicate diagnostic and therapeutic targets. In this study, we evaluated changes in the expression profile of Acanthamoeba proteins triggered by the invasive process, using an approach involving two-dimensional polyacrylamide gel electrophoresis (2DE PAGE), followed by mass spectrometry identification (ESI-IT-TOF LC-MSn). AK was induced by intrastromal inoculation in Wistar rats, using trophozoites from a T4 genotype, human case-derived A. castellanii strain under prolonged axenic culture. Cultures re-isolated from the lesions after two successive passages in the animals were used as biological triplicate for proteomic experiments. Analysis of the protein profile comparing long-term and re-isolated cultures indicated 62 significant spots, from which 27 proteins could be identified in the Acanthamoeba proteome database. Five of them (Serpin, Carboxypeptidase A1, Hypothetical protein, Calponin domain-containing protein, aldo/keto reductase) were exclusively found in the re-isolated trophozoites. Our analysis also revealed that a concerted modulation of several biochemical pathways is triggered when A. castellanii switches from a free-living style to a parasitic mode, including energetic metabolism, proteolytic activity, control of gene expression, protein degradation and methylation of DNA, which may be also involved in gain of virulence in an animal model of AK.A ceratite amebiana (CA) é uma infecção com risco de visão caracterizada por uma inflamação grave da córnea, causada pelo protozoário de vida livre do gênero Acanthamoeba. A identificação de proteínas amebianas envolvidas na patogênese da AK pode ajudar a elucidar os mecanismos moleculares da infecção e contribuir para indicar alvos diagnósticos e terapêuticos. Neste estudo, avaliamos as mudanças no perfil de expressão de proteínas de Acanthamoeba desencadeadas pelo processo invasivo, usando uma abordagem envolvendo eletroforese bidimensional em gel de poliacrilamida (2DE PAGE), seguida de identificação por espectrometria de massas (ESI-IT-TOF LC-MSn). AK foi induzida por inoculação intraestromal em ratos Wistar, usando trofozoítos de um genótipo T4, cepa de A. castellanii derivada de caso humano sob cultura axênica prolongada. Culturas reisoladas das lesões após duas passagens sucessivas nos animais foram utilizadas como triplicata biológica para experimentos proteômicos. A análise do perfil proteico comparando culturas de longo prazo e reisoladas indicou 62 pontos significativos, dos quais 27 proteínas puderam ser identificadas no banco de dados de proteomas de Acanthamoeba. Cinco deles (Serpina, Carboxipeptidase A1, Proteína hipotética, Proteína contendo o domínio Calponina, aldo/ceto redutase) foram encontrados exclusivamente nos trofozoítos reisolados. Nossa análise também revelou que uma modulação coordenada de várias vias bioquímicas é desencadeada quando A. castellanii muda de um estilo de vida livre para um modo parasitário, incluindo metabolismo energético, atividade proteolítica, controle da expressão gênica, degradação de proteínas e metilação do DNA, que também pode estar envolvido no ganho de virulência em um modelo animal de AK.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorFINEP - Financiadora de Estudos e Projetos, Financiadora de Estudos e ProjetosengUniversidade Federal de Minas GeraisUFMGBrasilFAR - DEPARTAMENTO DE ANÁLISES CLÍNICAS E TOXICOLÓGICASICB - DEPARTAMENTO DE MICROBIOLOGIAExperimental ParasitologyAcanthamoebaCeratiteProteômicaAcanthamoebaAmoebic keratitis rat modelProteomicsDifferential expression of Acanthamoeba castellanii proteins during amoebic keratitis in ratsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttps://www.sciencedirect.com/science/article/pii/S0014489420305853Ana Carolina Carvalho SilvaCamila Henriques CoelhoCecília Cirelli dos Santos FerreiraFrederico Crepaldi NascimentoIsabela Aurora Rodrigues ChagasCinthia Furst Leroy Gomes BueloniDaniel Carvalho PimentaJuliano Simões de ToledoAna Paula Salles Moura FernandesAdriana Oliveira Costaapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSELicense.txtLicense.txttext/plain; charset=utf-82042https://repositorio.ufmg.br/bitstream/1843/54632/1/License.txtfa505098d172de0bc8864fc1287ffe22MD51ORIGINALDifferential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats.pdfDifferential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats.pdfapplication/pdf5254070https://repositorio.ufmg.br/bitstream/1843/54632/2/Differential%20expression%20of%20Acanthamoeba%20castellanii%20proteins%20during%20amoebic%20keratitis%20in%20rats.pdfc4175c5cd60de4f1d86de05d11fb1e01MD521843/546322023-06-07 16:55:52.068oai:repositorio.ufmg.br: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Repositório de PublicaçõesPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-06-07T19:55:52Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false |
dc.title.pt_BR.fl_str_mv |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
title |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
spellingShingle |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats Ana Carolina Carvalho Silva Acanthamoeba Amoebic keratitis rat model Proteomics Acanthamoeba Ceratite Proteômica |
title_short |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
title_full |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
title_fullStr |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
title_full_unstemmed |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
title_sort |
Differential expression of Acanthamoeba castellanii proteins during amoebic keratitis in rats |
author |
Ana Carolina Carvalho Silva |
author_facet |
Ana Carolina Carvalho Silva Camila Henriques Coelho Cecília Cirelli dos Santos Ferreira Frederico Crepaldi Nascimento Isabela Aurora Rodrigues Chagas Cinthia Furst Leroy Gomes Bueloni Daniel Carvalho Pimenta Juliano Simões de Toledo Ana Paula Salles Moura Fernandes Adriana Oliveira Costa |
author_role |
author |
author2 |
Camila Henriques Coelho Cecília Cirelli dos Santos Ferreira Frederico Crepaldi Nascimento Isabela Aurora Rodrigues Chagas Cinthia Furst Leroy Gomes Bueloni Daniel Carvalho Pimenta Juliano Simões de Toledo Ana Paula Salles Moura Fernandes Adriana Oliveira Costa |
author2_role |
author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Ana Carolina Carvalho Silva Camila Henriques Coelho Cecília Cirelli dos Santos Ferreira Frederico Crepaldi Nascimento Isabela Aurora Rodrigues Chagas Cinthia Furst Leroy Gomes Bueloni Daniel Carvalho Pimenta Juliano Simões de Toledo Ana Paula Salles Moura Fernandes Adriana Oliveira Costa |
dc.subject.por.fl_str_mv |
Acanthamoeba Amoebic keratitis rat model Proteomics |
topic |
Acanthamoeba Amoebic keratitis rat model Proteomics Acanthamoeba Ceratite Proteômica |
dc.subject.other.pt_BR.fl_str_mv |
Acanthamoeba Ceratite Proteômica |
description |
Amoebic keratitis (AK) is a sight-threatening infection characterized by a severe inflammation of the cornea, caused by the free-living protozoan of the genus Acanthamoeba. Identification of amoebic proteins involved in AK pathogenesis may help to elucidate molecular mechanisms of infection and contribute to indicate diagnostic and therapeutic targets. In this study, we evaluated changes in the expression profile of Acanthamoeba proteins triggered by the invasive process, using an approach involving two-dimensional polyacrylamide gel electrophoresis (2DE PAGE), followed by mass spectrometry identification (ESI-IT-TOF LC-MSn). AK was induced by intrastromal inoculation in Wistar rats, using trophozoites from a T4 genotype, human case-derived A. castellanii strain under prolonged axenic culture. Cultures re-isolated from the lesions after two successive passages in the animals were used as biological triplicate for proteomic experiments. Analysis of the protein profile comparing long-term and re-isolated cultures indicated 62 significant spots, from which 27 proteins could be identified in the Acanthamoeba proteome database. Five of them (Serpin, Carboxypeptidase A1, Hypothetical protein, Calponin domain-containing protein, aldo/keto reductase) were exclusively found in the re-isolated trophozoites. Our analysis also revealed that a concerted modulation of several biochemical pathways is triggered when A. castellanii switches from a free-living style to a parasitic mode, including energetic metabolism, proteolytic activity, control of gene expression, protein degradation and methylation of DNA, which may be also involved in gain of virulence in an animal model of AK. |
publishDate |
2021 |
dc.date.issued.fl_str_mv |
2021-02 |
dc.date.accessioned.fl_str_mv |
2023-06-07T11:29:10Z |
dc.date.available.fl_str_mv |
2023-06-07T11:29:10Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1843/54632 |
dc.identifier.doi.pt_BR.fl_str_mv |
https://doi.org/10.1016/j.exppara.2020.108060 |
dc.identifier.issn.pt_BR.fl_str_mv |
0753-3322 |
url |
https://doi.org/10.1016/j.exppara.2020.108060 http://hdl.handle.net/1843/54632 |
identifier_str_mv |
0753-3322 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Experimental Parasitology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
dc.publisher.initials.fl_str_mv |
UFMG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
FAR - DEPARTAMENTO DE ANÁLISES CLÍNICAS E TOXICOLÓGICAS ICB - DEPARTAMENTO DE MICROBIOLOGIA |
publisher.none.fl_str_mv |
Universidade Federal de Minas Gerais |
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reponame:Repositório Institucional da UFMG instname:Universidade Federal de Minas Gerais (UFMG) instacron:UFMG |
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Universidade Federal de Minas Gerais (UFMG) |
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UFMG |
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Repositório Institucional da UFMG |
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