Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection
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 ch...
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| Format: | Article |
| Language: | English |
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Elsevier
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| Series: | Brazilian Journal of Infectious Diseases |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702008000200003&lng=en&tlng=en |
| _version_ | 1818335460390338560 |
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| author | Priscila F.R. Palma Giovana L. Baggio Celso Spada Renata da Silva Silvia Inês A.C.P. Ferreira Aricio Treitinger |
| author_facet | Priscila F.R. Palma Giovana L. Baggio Celso Spada Renata da Silva Silvia Inês A.C.P. Ferreira Aricio Treitinger |
| author_sort | Priscila F.R. Palma |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-13T14:23:47Z |
| format | Article |
| id | doaj.art-496f6260615f4cbd80aded051c213882 |
| institution | Directory Open Access Journal |
| issn | 1678-4391 |
| language | English |
| last_indexed | 2024-12-13T14:23:47Z |
| publisher | Elsevier |
| record_format | Article |
| series | Brazilian Journal of Infectious Diseases |
| spelling | doaj.art-496f6260615f4cbd80aded051c2138822022-12-21T23:42:00ZengElsevierBrazilian Journal of Infectious Diseases1678-439112210811410.1590/S1413-86702008000200003S1413-86702008000200003Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infectionPriscila F.R. Palma0Giovana L. Baggio1Celso Spada2Renata da Silva3Silvia Inês A.C.P. Ferreira4Aricio Treitinger5Universidade Federal de Santa CatarinaUniversidade Federal de São PauloUniversidade Federal de Santa CatarinaSanta Luzia Medical LaboratorySanta Luzia Medical LaboratoryUniversidade Federal de Santa CatarinaEvaluation 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702008000200003&lng=en&tlng=enApoptosisflow cytometryannexin Vpropidium iodidecalcein-AMethidium homodimer |
| spellingShingle | Priscila F.R. Palma Giovana L. Baggio Celso Spada Renata da Silva Silvia Inês A.C.P. Ferreira Aricio Treitinger Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection Brazilian Journal of Infectious Diseases Apoptosis flow cytometry annexin V propidium iodide calcein-AM ethidium homodimer |
| title | 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_short | 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 |
| topic | Apoptosis flow cytometry annexin V propidium iodide calcein-AM ethidium homodimer |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702008000200003&lng=en&tlng=en |
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