Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response

Detalhes bibliográficos
Autor(a) principal: Silva, Icaro Bonyek
Data de Publicação: 2021
Outros Autores: Nunes, Sara, Bastos, Rana, Lima, Reinan, Barbosa, Leilane, Grimaldi, Gabriela, Rocha, Vinicius, Soares, Milena Botelho Pereira, Veras, Patrícia Sampaio Tavares, Menezes, Juliana de, Brodskyn, Cláudia Ida, Tavares, Natalia
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/50153
Resumo: Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) under Grant number PET0009/2016 and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) under Finance Code 001.
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spelling Silva, Icaro BonyekNunes, SaraBastos, RanaLima, ReinanBarbosa, LeilaneGrimaldi, GabrielaRocha, ViniciusSoares, Milena Botelho PereiraVeras, Patrícia Sampaio TavaresMenezes, Juliana deBrodskyn, Cláudia IdaTavares, Natalia2021-12-03T13:31:28Z2021-12-03T13:31:28Z2021SILVA, Icaro Bonyek et al. Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response. Journal of Visualized Experiments, 2021.1940-087Xhttps://www.arca.fiocruz.br/handle/icict/5015310.3791/62555 (2021).Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) under Grant number PET0009/2016 and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) under Finance Code 001.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Senai Institute for Innovation in Advanced Health Systems. National Service for Industrial Learning. Integrated Manufacturing and Technology Campus. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Senai Institute for Innovation in Advanced Health Systems. National Service for Industrial Learning. Integrated Manufacturing and Technology Campus. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / National Institute of Science and Technology. Research Institute in Immunology. Salvador, BA, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil / National Institute of Science and Technology. Research Institute in Immunology. Salvador, BA, Brazil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Macrophages are multifunctional cells essential to the immune system function, and the primary host cell in Leishmania braziliensis (Lb) infection. These cells are specialized in microorganism recognition and phagocytosis, but also activate other immune cells and present antigens, as well as promote inflammation and tissue repair. Here, we describe a protocol to obtain mononuclear cells from peripheral blood (PBMC) of healthy donors to separate monocytes that then differentiate into macrophages. These cells can then be infected in vitro at different Lb concentrations to evaluate the ability to control infection, as well as evaluate host cell immune response, which can be measured by several methods. PBMCs were first isolated by centrifuging with Ficoll-Hypaque gradient and then plated to allow monocytes to adhere to culture plates; non-adherent cells were removed by washing. Next, adherent cells were cultured with macrophage-colony stimulating factor (M-CSF) for 7 days to induce macrophage differentiation. We suggest plating 2 x 106 cells per well on 24-well plates in order to obtain 2 x 105 macrophages. Fully differentiated macrophages can then be infected with Lb for 4 or 24 hours. This protocol results in a significant percentage of infected cells, which can be assessed by optical or fluorescence microscopy. In addition to infection index, parasite load can be measured by counting the numbers of parasites inside each cell. Further molecular and functional assays can also be performed in culture supernatants or within the macrophages themselves, which allows this protocol to be applied in a variety of contexts and also adapted to other intracellular parasite species. IntroductionengMyJove CorporationLeishmania braziliensisMacrófagosMonócitosMicroscopia de FluorescênciaLeishmania braziliensisMacrophagesMonocytesMicroscopy, FluorescenceObtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Responseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83097https://www.arca.fiocruz.br/bitstream/icict/50153/1/license.txt36b51ef91c52b5338d9d29ba0cc807bcMD51ORIGINALSilva, Icaro Bonyek Obtainment-macrophages-from....pdfSilva, Icaro Bonyek Obtainment-macrophages-from....pdfapplication/pdf569843https://www.arca.fiocruz.br/bitstream/icict/50153/2/Silva%2c%20Icaro%20Bonyek%20Obtainment-macrophages-from....pdf1c22ebcbed98d2edbc1ef8838b461fffMD52icict/501532023-03-15 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dc.title.pt_BR.fl_str_mv Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
title Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
spellingShingle Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
Silva, Icaro Bonyek
Leishmania braziliensis
Macrófagos
Monócitos
Microscopia de Fluorescência
Leishmania braziliensis
Macrophages
Monocytes
Microscopy, Fluorescence
title_short Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
title_full Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
title_fullStr Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
title_full_unstemmed Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
title_sort Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response
author Silva, Icaro Bonyek
author_facet Silva, Icaro Bonyek
Nunes, Sara
Bastos, Rana
Lima, Reinan
Barbosa, Leilane
Grimaldi, Gabriela
Rocha, Vinicius
Soares, Milena Botelho Pereira
Veras, Patrícia Sampaio Tavares
Menezes, Juliana de
Brodskyn, Cláudia Ida
Tavares, Natalia
author_role author
author2 Nunes, Sara
Bastos, Rana
Lima, Reinan
Barbosa, Leilane
Grimaldi, Gabriela
Rocha, Vinicius
Soares, Milena Botelho Pereira
Veras, Patrícia Sampaio Tavares
Menezes, Juliana de
Brodskyn, Cláudia Ida
Tavares, Natalia
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Silva, Icaro Bonyek
Nunes, Sara
Bastos, Rana
Lima, Reinan
Barbosa, Leilane
Grimaldi, Gabriela
Rocha, Vinicius
Soares, Milena Botelho Pereira
Veras, Patrícia Sampaio Tavares
Menezes, Juliana de
Brodskyn, Cláudia Ida
Tavares, Natalia
dc.subject.other.pt_BR.fl_str_mv Leishmania braziliensis
Macrófagos
Monócitos
Microscopia de Fluorescência
topic Leishmania braziliensis
Macrófagos
Monócitos
Microscopia de Fluorescência
Leishmania braziliensis
Macrophages
Monocytes
Microscopy, Fluorescence
dc.subject.en.pt_BR.fl_str_mv Leishmania braziliensis
Macrophages
Monocytes
Microscopy, Fluorescence
description Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB) under Grant number PET0009/2016 and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) under Finance Code 001.
publishDate 2021
dc.date.accessioned.fl_str_mv 2021-12-03T13:31:28Z
dc.date.available.fl_str_mv 2021-12-03T13:31:28Z
dc.date.issued.fl_str_mv 2021
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.citation.fl_str_mv SILVA, Icaro Bonyek et al. Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response. Journal of Visualized Experiments, 2021.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/50153
dc.identifier.issn.pt_BR.fl_str_mv 1940-087X
dc.identifier.eissn.none.fl_str_mv 10.3791/62555 (2021).
identifier_str_mv SILVA, Icaro Bonyek et al. Obtainment of Macrophages from Human Monocytes to Assess Leishmania braziliensis Infection Rate and Innate Host Immune Response. Journal of Visualized Experiments, 2021.
1940-087X
10.3791/62555 (2021).
url https://www.arca.fiocruz.br/handle/icict/50153
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MyJove Corporation
publisher.none.fl_str_mv MyJove Corporation
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/50153/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/50153/2/Silva%2c%20Icaro%20Bonyek%20Obtainment-macrophages-from....pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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