Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis

Author(s): Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, et al.

Abstract

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.

Similar Articles

Folic acid-CdTe quantum dot conjugates and their applications for cancer cell targeting

Author(s): Suriamoorthy P, Zhang X, Hao G, Joly AG, Singh S, et al.

A method to predict breast cancer stage using Medicare claims

Author(s): Smith GL, Shih YC, Giordano SH, Smith BD, Buchholz TA

Toxic potential of materials at the nanolevel

Author(s): Nel A, Xia T, M├Ądler L, Li N

From nanotechnology to nanomedicine: applications to cancer research

Author(s): Seigneuric R, Markey L, Nuyten DS, Dubernet C, Evelo CT, et al.

Superparamagnetic iron oxide nanoparticle probes for molecular imaging

Author(s): Thorek DL, Chen AK, Czupryna J, Tsourkas A

Freeze-drying of nanoparticles: formulation, process and storage considerations

Author(s): Abdelwahed W, Degobert G, Stainmesse S, Fessi H

Synthesis of gold nanotriangles and silver nanoparticles using Aloe vera plant extract

Author(s): Chandran SP, Chaudhary M, Pasricha R, Ahmad A, Sastry M

Facile green synthesis of variable metallic gold nanoparticle using Padinagymnospora, a brown marine macroalga

Author(s): Singh M, Kalaivani R, Manikandan S, Sangeetha N, Kumaraguru AK

Gold nanoparticles: From nanomedicine to nanosensing

Author(s): Chen PC, Mwakwari SC, Oyelere AK

Near-infrared resonant nanoshells for combined optical imaging and photothermal cancer therapy

Author(s): Gobin AM, Lee MH, Halas NJ, James WD, Drezek RA, et al.

Biological applications of gold nanoparticles

Author(s): Sperling RA, Rivera Gil P, Zhang F, Zanella M, Parak WJ

Gold nanoparticles for biology and medicine

Author(s): Giljohann DA, Seferos DS, Daniel WL, Massich MD, Patel PC, et al.

Reperfusion induces myocardial apoptotic cell death

Author(s): Zhao ZQ, Nakamura M, Wang NP, Wilcox JN, Shearer S, et al.

Asbestos causes apoptosis in alveolar epithelial cells: role of iron-induced free radicals

Author(s): Aljandali A, Pollack H, Yeldandi A, Li Y, Weitzman SA, et al.

Distinct cytotoxic mechanisms of pristine versus hydroxylated fullerene

Author(s): Isakovic A, Markovic Z, Todorovic-Markovic B, Nikolic N, Vranjes-Djuric S, et al.

In vitro testing of the potential for orthopedic bone cements to cause apoptosis of osteoblast-like cells

Author(s): Ciapetti G, Granchi D, Savarino L, Cenni E, Magrini E, et al.