Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/4349 http://dx.doi.org/10.1590/S1413-86702008000200003 |
Resumo: | Evaluation of apoptosis by flow cytometry is generally accomplished by methods that use annexin V-FITC as vital dye, which access phosphatidylserine exposed on the external membrane at the beginning of this process. In addition, the concomitant use of propidium iodide makes possible to verify the characteristic nuclear alterations in the late stages of apoptosis, as a consequence of the increase in membrane permeability. On the other hand, the use of calcein-AM in association with ethidium homodimer (EthD-1) allows the evaluation of cell apoptosis through detection of esterase activity and cellular membrane physical and chemical alterations. The aim of this study was to compare the sensibility of calcein-AM and EthD-1 with annexin V-FITC and propidium iodide for early apoptosis evaluation in peripheral blood mononuclear cell culture, obtained from HIV-infected patients. Apoptosis and cellular viability were detected and quantified by flow cytometry after 24 and 48 hours incubation times. Our results showed that calcein-AM/EthD-1 was more sensitive for apoptotic cell quantification in both incubation times than annexin V-FITC/propidium iodide (mean of 46.95% ± 3.56, p < 0.0001, for 24 hours and mean of 37.67% ± 2.47, p < 0.0014 for 48 hours), besides allowing to clearly define viable, apoptotic and dead cell populations. |
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Palma, Priscila F.r.Baggio-Zappia, Giovana Lotici [UNIFESP]Spada, CelsoSilva, Renata DaFerreira, Silvia Inês A.c.p.Treitinger, AricioFederal University of Santa Catarina Department of Clinical AnalysisUniversidade Federal de São Paulo (UNIFESP)Santa Luzia Medical Laboratory2015-06-14T13:38:31Z2015-06-14T13:38:31Z2008-04-01Brazilian Journal of Infectious Diseases. Brazilian Society of Infectious Diseases, v. 12, n. 2, p. 108-114, 2008.1413-8670http://repositorio.unifesp.br/handle/11600/4349http://dx.doi.org/10.1590/S1413-86702008000200003S1413-86702008000200003.pdfS1413-8670200800020000310.1590/S1413-86702008000200003WOS:000257897000003Evaluation of apoptosis by flow cytometry is generally accomplished by methods that use annexin V-FITC as vital dye, which access phosphatidylserine exposed on the external membrane at the beginning of this process. In addition, the concomitant use of propidium iodide makes possible to verify the characteristic nuclear alterations in the late stages of apoptosis, as a consequence of the increase in membrane permeability. On the other hand, the use of calcein-AM in association with ethidium homodimer (EthD-1) allows the evaluation of cell apoptosis through detection of esterase activity and cellular membrane physical and chemical alterations. The aim of this study was to compare the sensibility of calcein-AM and EthD-1 with annexin V-FITC and propidium iodide for early apoptosis evaluation in peripheral blood mononuclear cell culture, obtained from HIV-infected patients. Apoptosis and cellular viability were detected and quantified by flow cytometry after 24 and 48 hours incubation times. Our results showed that calcein-AM/EthD-1 was more sensitive for apoptotic cell quantification in both incubation times than annexin V-FITC/propidium iodide (mean of 46.95% ± 3.56, p < 0.0001, for 24 hours and mean of 37.67% ± 2.47, p < 0.0014 for 48 hours), besides allowing to clearly define viable, apoptotic and dead cell populations.Federal University of Santa Catarina Department of Clinical AnalysisFederal University of São Paulo Division of Infectious DiseasesSanta Luzia Medical LaboratoryUNIFESP, Division of Infectious DiseasesSciELO108-114engBrazilian Society of Infectious DiseasesBrazilian Journal of Infectious DiseasesApoptosisflow cytometryannexin Vpropidium iodidecalcein-AMethidium homodimerEvaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infectioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALS1413-86702008000200003.pdfapplication/pdf206419${dspace.ui.url}/bitstream/11600/4349/1/S1413-86702008000200003.pdfa4dc077875a242c648c5c0aabff4564cMD51open accessTEXTS1413-86702008000200003.pdf.txtS1413-86702008000200003.pdf.txtExtracted texttext/plain30796${dspace.ui.url}/bitstream/11600/4349/2/S1413-86702008000200003.pdf.txt5a389b9083dd1071c17501e0de897b33MD52open access11600/43492021-10-04 21:20:47.978open accessoai:repositorio.unifesp.br:11600/4349Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652021-10-05T00:20:47Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
title |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
spellingShingle |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection Palma, Priscila F.r. Apoptosis flow cytometry annexin V propidium iodide calcein-AM ethidium homodimer |
title_short |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
title_full |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
title_fullStr |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
title_full_unstemmed |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
title_sort |
Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection |
author |
Palma, Priscila F.r. |
author_facet |
Palma, Priscila F.r. Baggio-Zappia, Giovana Lotici [UNIFESP] Spada, Celso Silva, Renata Da Ferreira, Silvia Inês A.c.p. Treitinger, Aricio |
author_role |
author |
author2 |
Baggio-Zappia, Giovana Lotici [UNIFESP] Spada, Celso Silva, Renata Da Ferreira, Silvia Inês A.c.p. Treitinger, Aricio |
author2_role |
author author author author author |
dc.contributor.institution.none.fl_str_mv |
Federal University of Santa Catarina Department of Clinical Analysis Universidade Federal de São Paulo (UNIFESP) Santa Luzia Medical Laboratory |
dc.contributor.author.fl_str_mv |
Palma, Priscila F.r. Baggio-Zappia, Giovana Lotici [UNIFESP] Spada, Celso Silva, Renata Da Ferreira, Silvia Inês A.c.p. Treitinger, Aricio |
dc.subject.eng.fl_str_mv |
Apoptosis flow cytometry annexin V propidium iodide calcein-AM ethidium homodimer |
topic |
Apoptosis flow cytometry annexin V propidium iodide calcein-AM ethidium homodimer |
description |
Evaluation of apoptosis by flow cytometry is generally accomplished by methods that use annexin V-FITC as vital dye, which access phosphatidylserine exposed on the external membrane at the beginning of this process. In addition, the concomitant use of propidium iodide makes possible to verify the characteristic nuclear alterations in the late stages of apoptosis, as a consequence of the increase in membrane permeability. On the other hand, the use of calcein-AM in association with ethidium homodimer (EthD-1) allows the evaluation of cell apoptosis through detection of esterase activity and cellular membrane physical and chemical alterations. The aim of this study was to compare the sensibility of calcein-AM and EthD-1 with annexin V-FITC and propidium iodide for early apoptosis evaluation in peripheral blood mononuclear cell culture, obtained from HIV-infected patients. Apoptosis and cellular viability were detected and quantified by flow cytometry after 24 and 48 hours incubation times. Our results showed that calcein-AM/EthD-1 was more sensitive for apoptotic cell quantification in both incubation times than annexin V-FITC/propidium iodide (mean of 46.95% ± 3.56, p < 0.0001, for 24 hours and mean of 37.67% ± 2.47, p < 0.0014 for 48 hours), besides allowing to clearly define viable, apoptotic and dead cell populations. |
publishDate |
2008 |
dc.date.issued.fl_str_mv |
2008-04-01 |
dc.date.accessioned.fl_str_mv |
2015-06-14T13:38:31Z |
dc.date.available.fl_str_mv |
2015-06-14T13:38:31Z |
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 |
Brazilian Journal of Infectious Diseases. Brazilian Society of Infectious Diseases, v. 12, n. 2, p. 108-114, 2008. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/4349 http://dx.doi.org/10.1590/S1413-86702008000200003 |
dc.identifier.issn.none.fl_str_mv |
1413-8670 |
dc.identifier.file.none.fl_str_mv |
S1413-86702008000200003.pdf |
dc.identifier.scielo.none.fl_str_mv |
S1413-86702008000200003 |
dc.identifier.doi.none.fl_str_mv |
10.1590/S1413-86702008000200003 |
dc.identifier.wos.none.fl_str_mv |
WOS:000257897000003 |
identifier_str_mv |
Brazilian Journal of Infectious Diseases. Brazilian Society of Infectious Diseases, v. 12, n. 2, p. 108-114, 2008. 1413-8670 S1413-86702008000200003.pdf S1413-86702008000200003 10.1590/S1413-86702008000200003 WOS:000257897000003 |
url |
http://repositorio.unifesp.br/handle/11600/4349 http://dx.doi.org/10.1590/S1413-86702008000200003 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Brazilian Journal of Infectious Diseases |
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info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
108-114 |
dc.publisher.none.fl_str_mv |
Brazilian Society of Infectious Diseases |
publisher.none.fl_str_mv |
Brazilian Society of Infectious Diseases |
dc.source.none.fl_str_mv |
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