Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis.
نویسندگان
چکیده
Apoptosis, or programmed cell death, is a general mechanism for removal of unwanted cells from the immune system. It is characterized by chromatin condensation, a reduction in cell volume, and endonuclease cleavage of DNA into oligonucleosomal length fragments. Apoptosis is also accompanied by a loss of membrane phospholipid asymmetry, resulting in the exposure of phosphatidylserine at the surface of the cell. Expression of phosphatidylserine at the cell surface plays an important role in the recognition and removal of apoptotic cells by macrophages. Here we describe a new method for the detection of apoptotic cells by flow cytometry, using the binding of fluorescein isothiocyanate-labeled annexin V to phosphatidylserine. When Burkitt lymphoma cell lines and freshly isolated germinal center B cells are cultured under apoptosis inducing conditions, all cells showing chromatin condensation strongly stain with annexin V, whereas normal cells are annexin V negative. Moreover, DNA fragmentation is only found in the annexin V-positive cells. The nonvital dye ethidium bromide was found to stain a subpopulation of the annexin V-positive apoptotic cells, increasing with time. Our results indicate that the phase in apoptosis that is characterized by chromatin condensation coincides with phosphatidylserine exposure. Importantly, it precedes membrane damage that might lead to release from the cells of enzymes that are harmful to the surrounding tissues. Annexin V may prove important in further unravelling the regulation of apoptosis.
منابع مشابه
Annexin V for Flow Cytometric Detection of Phosphatidylserine Expression on B Cells Undergoing Apoptosis
Apoptosis, or programmed cell death, is a general mechanism for removal of unwanted cells from the immune system. It is characterized by chromatin condensation, a reduction in cell volume, and endonuclease cleavage of DNA into oligonucleosomal length fragments. Apoptosis is also accompanied by a loss of membrane phospholipid asymmetry, resulting in the exposure of phosphatidylserine at the surf...
متن کامل[Annexin V for flow cytometric detection of phosphatidylserine expression on lymphoma cells undergoing apoptosis].
OBJECTIVE To quantitatively analyze apoptotic and secondary necrotic cells under apoptosis conditions. METHODS The cells of Burkitt lymphoma cell line Raji were incubated with 1.0 mumol/L dexamethasone (DEX) for 2, 4 and 8 h, then stained with Annexin V-FITC (fluorescein isothiocyanate conjugated) which was used to detect the exposure of phosphatidylserine (PS) on the out membrane resulting f...
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ورودعنوان ژورنال:
- Blood
دوره 84 5 شماره
صفحات -
تاریخ انتشار 1994