Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection

Evaluation of annexin V and Calcein-AM as markers of mononuclear cell apoptosis during human immunodeficiency virus infection

Author Palma, Priscila F.r. Google Scholar
Baggio-Zappia, Giovana Lotici Autor UNIFESP Google Scholar
Spada, Celso Google Scholar
Silva, Renata Da Google Scholar
Ferreira, Silvia Inês A.c.p. Google Scholar
Treitinger, Aricio Google Scholar
Institution Federal University of Santa Catarina Department of Clinical Analysis
Universidade Federal de São Paulo (UNIFESP)
Santa Luzia Medical Laboratory
Abstract 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.
Keywords Apoptosis
flow cytometry
annexin V
propidium iodide
calcein-AM
ethidium homodimer
Language English
Date 2008-04-01
Published in Brazilian Journal of Infectious Diseases. Brazilian Society of Infectious Diseases, v. 12, n. 2, p. 108-114, 2008.
ISSN 1413-8670 (Sherpa/Romeo, impact factor)
Publisher Brazilian Society of Infectious Diseases
Extent 108-114
Origin http://dx.doi.org/10.1590/S1413-86702008000200003
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000257897000003
SciELO ID S1413-86702008000200003 (statistics in SciELO)
URI http://repositorio.unifesp.br/handle/11600/4349

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