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Previous Article | Table of Contents | Next Article 
Annexin V for flow cytometric detection of phosphatidylserine expression on
B cells undergoing apoptosis
G Koopman, CP Reutelingsperger, GA Kuijten, RM Keehnen, ST Pals and MH van Oers
Department of Pathology, Academic Medical Center, Amsterdam, The
Netherlands.
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.
Volume 84,
Issue 5,
pp. 1415-1420,
09/01/1994
Copyright © 1994 by The American Society of Hematology

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M. A. Salako, M. J. Carter, and G. E. N. Kass
Coxsackievirus Protein 2BC Blocks Host Cell Apoptosis by Inhibiting Caspase-3
J. Biol. Chem.,
June 16, 2006;
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[Abstract]
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W. Xu, A. Roos, N. Schlagwein, A. M. Woltman, M. R. Daha, and C. van Kooten
IL-10-producing macrophages preferentially clear early apoptotic cells
Blood,
June 15, 2006;
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[Abstract]
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J.-B. Park, I.-C. Park, S.-J. Park, H.-O. Jin, J.-K. Lee, and K. D. Riew
Anti-Apoptotic Effects of Caspase Inhibitors on Rat Intervertebral Disc Cells
J. Bone Joint Surg. Am.,
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[Abstract]
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S. K. Patnaik, B. Potvin, S. Carlsson, D. Sturm, H. Leffler, and P. Stanley
Complex N-glycans are the major ligands for galectin-1, -3, and -8 on Chinese hamster ovary cells
Glycobiology,
April 1, 2006;
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[Abstract]
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T. Ono, K.-S. Park, M. Ueta, T. Iida, and T. Honda
Identification of Proteins Secreted via Vibrio parahaemolyticus Type III Secretion System 1
Infect. Immun.,
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[Abstract]
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M. F. Corsten, L. Hofstra, J. Narula, and C. P.M. Reutelingsperger
Counting Heads in the War against Cancer: Defining the Role of Annexin A5 Imaging in Cancer Treatment and Surveillance
Cancer Res.,
February 1, 2006;
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M. Bungum, L. Bungum, and P. Humaidan
A prospective study, using sibling oocytes, examining the effect of 30 seconds versus 90 minutes gamete co-incubation in IVF
Hum. Reprod.,
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N. Berkova, S. Lair-Fulleringer, F. Femenia, D. Huet, M.-C. Wagner, K. Gorna, F. Tournier, O. Ibrahim-Granet, J. Guillot, R. Chermette, et al.
Aspergillus fumigatus conidia inhibit tumour necrosis factor- or staurosporine-induced apoptosis in epithelial cells
Int. Immunol.,
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H. H. Boersma, B. L.J.H. Kietselaer, L. M.L. Stolk, A. Bennaghmouch, L. Hofstra, J. Narula, G. A.K. Heidendal, and C. P.M. Reutelingsperger
Past, Present, and Future of Annexin A5: From Protein Discovery to Clinical Applications
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T. Ohara, T. Itoh, and M. Takahashi
Immunosuppression by Morphine-Induced Lymphocyte Apoptosis: Is It a Real Issue?
Anesth. Analg.,
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R. A. Barrington, M. Zhang, X. Zhong, H. Jonsson, N. Holodick, A. Cherukuri, S. K. Pierce, T. L. Rothstein, and M. C. Carroll
CD21/CD19 Coreceptor Signaling Promotes B Cell Survival during Primary Immune Responses
J. Immunol.,
September 1, 2005;
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R. A. Mesa, D. Loegering, H. L. Powell, K. Flatten, S. J. H. Arlander, N. T. Dai, M. P. Heldebrant, B. T. Vroman, B. D. Smith, J. E. Karp, et al.
Heat shock protein 90 inhibition sensitizes acute myelogenous leukemia cells to cytarabine
Blood,
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L. M. Lashinger, K. Zhu, S. A. Williams, M. Shrader, C. P.N. Dinney, and D. J. McConkey
Bortezomib Abolishes Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Resistance via a p21-Dependent Mechanism in Human Bladder and Prostate Cancer Cells
Cancer Res.,
June 1, 2005;
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J. Quadrilatero and L. Hoffman-Goetz
N-acetyl-l-cysteine protects intestinal lymphocytes from apoptotic death after acute exercise in adrenalectomized mice
Am J Physiol Regulatory Integrative Comp Physiol,
June 1, 2005;
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V. Pinho, D. G. Souza, M. M. Barsante, F. P. Hamer, M. S. De Freitas, A. G. Rossi, and M. M. Teixeira
Phosphoinositide-3 kinases critically regulate the recruitment and survival of eosinophils in vivo: importance for the resolution of allergic inflammation
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J. S. Mader, J. Salsman, D. M. Conrad, and D. W. Hoskin
Bovine lactoferricin selectively induces apoptosis in human leukemia and carcinoma cell lines
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S. M. Jackson and J. D. Capra
IgH V-Region Sequence Does Not Predict the Survival Fate of Human Germinal Center B Cells
J. Immunol.,
March 1, 2005;
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T. D. Shanafelt, Y. K. Lee, N. D. Bone, A. K. Strege, V. L. Narayanan, E. A. Sausville, S. M. Geyer, S. H. Kaufmann, and N. E. Kay
Adaphostin-induced apoptosis in CLL B cells is associated with induction of oxidative stress and exhibits synergy with fludarabine
Blood,
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[Abstract]
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P. R. Hoffmann, J. A. Kench, A. Vondracek, E. Kruk, D. L. Daleke, M. Jordan, P. Marrack, P. M. Henson, and V. A. Fadok
Interaction between Phosphatidylserine and the Phosphatidylserine Receptor Inhibits Immune Responses In Vivo
J. Immunol.,
February 1, 2005;
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A. F. Valledor, L.-C. Hsu, S. Ogawa, D. Sawka-Verhelle, M. Karin, and C. K. Glass
Activation of liver X receptors and retinoid X receptors prevents bacterial-induced macrophage apoptosis
PNAS,
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[Abstract]
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H.-G. Kim, C. G. de Guzman, C. S. Swindle, C. V. Cotta, L. Gartland, E. W. Scott, and C. A. Klug
The ETS family transcription factor PU.1 is necessary for the maintenance of fetal liver hematopoietic stem cells
Blood,
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[Abstract]
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E. I. Schwartz, R. V. Intine, and R. J. Maraia
CK2 Is Responsible for Phosphorylation of Human La Protein Serine-366 and Can Modulate rpL37 5'-Terminal Oligopyrimidine mRNA Metabolism
Mol. Cell. Biol.,
November 1, 2004;
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N. Shah, R. J. Asch, A. S. Lysholm, and T. W. LeBien
Enhancement of stress-induced apoptosis in B-lineage cells by caspase-9 inhibitor
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S. K. Mak and D. Kultz
Gadd45 Proteins Induce G2/M Arrest and Modulate Apoptosis in Kidney Cells Exposed to Hyperosmotic Stress
J. Biol. Chem.,
September 10, 2004;
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A. J. Nauta, G. Castellano, W. Xu, A. M. Woltman, M. C. Borrias, M. R. Daha, C. van Kooten, and A. Roos
Opsonization with C1q and Mannose-Binding Lectin Targets Apoptotic Cells to Dendritic Cells
J. Immunol.,
September 1, 2004;
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C Gabler, N Blank, T Hieronymus, M Schiller, J H M Berden, J R Kalden, and H-M Lorenz
Extranuclear detection of histones and nucleosomes in activated human lymphoblasts as an early event in apoptosis
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M. C. A. da Silva, J.-M. Zahm, D. Gras, O. Bajolet, M. Abely, J. Hinnrasky, M. Milliot, M. C. de Assis, C. Hologne, N. Bonnet, et al.
Dynamic interaction between airway epithelial cells and Staphylococcus aureus
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G. Kuznetsov, M. J. Towle, H. Cheng, T. Kawamura, K. TenDyke, D. Liu, Y. Kishi, M. J. Yu, and B. A. Littlefield
Induction of Morphological and Biochemical Apoptosis following Prolonged Mitotic Blockage by Halichondrin B Macrocyclic Ketone Analog E7389
Cancer Res.,
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E. Oei, T. Kalb, P. Beuria, M. Allez, A. Nakazawa, M. Azuma, M. Timony, Z. Stuart, H. Chen, and K. Sperber
Accessory cell function of airway epithelial cells
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J. H. Jhamandas and D. MacTavish
Antagonist of the Amylin Receptor Blocks {beta}-Amyloid Toxicity in Rat Cholinergic Basal Forebrain Neurons
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K. S. Lang, S. Myssina, P. A. Lang, V. Tanneur, D. S. Kempe, A. F. Mack, S. M. Huber, T. Wieder, F. Lang, and C. Duranton
Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl-
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Y. Hisaka, M. Ieda, T. Nakamura, K.-i. Kosai, S. Ogawa, and K. Fukuda
Powerful and controllable angiogenesis by using gene-modified cells expressing human hepatocyte growth factor and thymidine kinase
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R. J. Menaker, P. J. M. Ceponis, and N. L. Jones
Helicobacter pylori Induces Apoptosis of Macrophages in Association with Alterations in the Mitochondrial Pathway
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D. Chatterjee, Y. Bai, Z. Wang, S. Beach, S. Mott, R. Roy, C. Braastad, Y. Sun, A. Mukhopadhyay, B. B. Aggarwal, et al.
RKIP Sensitizes Prostate and Breast Cancer Cells to Drug-induced Apoptosis
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J. C. Todt, B. Hu, and J. L. Curtis
The receptor tyrosine kinase MerTK activates phospholipase C {gamma}2 during recognition of apoptotic thymocytes by murine macrophages
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X.-B. Wang, B.-F. Zhao, Q. Zhao, J.-H. Piao, J. Liu, Q. Lin, and H.-L. Huang
A New Recombinant Single Chain Trispecific Antibody Recruits T Lymphocytes to Kill CEA (Carcinoma Embryonic Antigen) Positive Tumor Cells In Vitro Efficiently
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T. M. Scarabelli, A. Stephanou, E. Pasini, G. Gitti, P. Townsend, K. Lawrence, C. Chen-Scarabelli, L. Saravolatz, D. Latchman, R. Knight, et al.
Minocycline inhibits caspase activation and reactivation, increases the ratio of XIAP to smac/DIABLO, and reduces the mitochondrial leakage of cytochrome C and smac/DIABLO
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S. Varambally, Y. Bar-Dayan, J. Bayry, S. Lacroix-Desmazes, M. Horn, M. Sorel, Y. Bar-Dayan, G. Ruberti, M. D. Kazatchkine, and S. V. Kaveri
Natural human polyreactive IgM induce apoptosis of lymphoid cell lines and human peripheral blood mononuclear cells
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