|
|
 Previous Article | Table of Contents | Next Article 
Detection of altered membrane phospholipid asymmetry in subpopulations of
human red blood cells using fluorescently labeled annexin V
FA Kuypers, RA Lewis, M Hua, MA Schott, D Discher, JD Ernst and BH Lubin
Children's Hospital Oakland Research Institute, CA 94609, USA.
The phospholipids of the human red cell are distributed asymmetrically in
the bilayer of the red cell membrane. In certain pathologic states, such as
sickle cell anemia, phospholipid asymmetry is altered. Although several
methods can be used to measure phospholipid organization, small
organizational changes have been very difficult to assess. Moreover, these
methods fail to identify subpopulations of cells that have lost their
normal phospholipid asymmetry. Using fluorescently labeled annexin V in
flow cytometry and fluorescent microscopy, we were able to identify and
quantify red cells that had lost their phospholipid asymmetry in
populations as small as 1 million cells. Moreover, loss of phospholipid
organization in subpopulations as small as 0.1% of the total population
could be identified, and individual cells could be studied by fluorescent
microscopy. An excellent correlation was found between
fluorescence-activated cell sorter (FACS) analysis results using annexin V
to detect red cells with phosphatidylserine (PS) on their surface and a
PS-requiring prothrombinase assay using similar red cells. Cells that bound
fluorescein isothiocyanate (FITC)-labeled annexin V could be isolated from
the population using magnetic beads covered with an anti-FITC antibody.
Evaluation of blood samples from patients with sickle cell anemia under
oxygenated conditions demonstrated the presence of subpopulations of cells
that had lost phospholipid asymmetry. While only a few red cells were
labeled in normal control samples (0.21% +/- 0.12%, n = 8), significantly
increased (P < .001) annexin V labeling was observed in samples from
patients with sickle cell anemia (2.18% +/- 1.21%, n = 13). We conclude
that loss of phospholipid asymmetry may occur in small subpopulations of
red cells and that fluorescently labeled annexin V can be used to quantify
and isolate these cells.
Volume 87,
Issue 3,
pp. 1179-1187,
02/01/1996
Copyright © 1996 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
M. Sorice, P. Matarrese, A. Tinari, A. M. Giammarioli, T. Garofalo, V. Manganelli, L. Ciarlo, L. Gambardella, G. Maccari, M. Botta, et al.
Raft component GD3 associates with tubulin following CD95/Fas ligation
FASEB J,
October 1, 2009;
23(10):
3298 - 3308.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Ozkan, H. Oren, M. Tatli, H. Ates, A. Kumral, and N. Duman
Erythroid Apoptosis in Idiopathic Neonatal Jaundice
Pediatrics,
May 1, 2008;
121(5):
e1348 - e1351.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Johnson and M. J. Telen
Adhesion molecules and hydroxyurea in the pathophysiology of sickle cell disease
Haematologica,
April 1, 2008;
93(4):
481 - 485.
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. N. Y. Setty and S. G. Betal
Microvascular endothelial cells express a phosphatidylserine receptor: a functionally active receptor for phosphatidylserine-positive erythrocytes
Blood,
January 15, 2008;
111(2):
905 - 914.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Guchhait, S. K Dasgupta, A. Le, S. Yellapragada, J. A. Lopez, and P. Thiagarajan
Lactadherin mediates sickle cell adhesion to vascular endothelial cells in flowing blood
Haematologica,
September 1, 2007;
92(9):
1266 - 1267.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
V. L. Lew, N. Daw, Z. Etzion, T. Tiffert, A. Muoma, L. Vanagas, and R. M. Bookchin
Effects of age-dependent membrane transport changes on the homeostasis of senescent human red blood cells
Blood,
August 15, 2007;
110(4):
1334 - 1342.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. I. Ataga and N. S. Key
Hypercoagulability in Sickle Cell Disease: New Approaches to an Old Problem
Hematology,
January 1, 2007;
2007(1):
91 - 96.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Haynes Jr., B. Obiako, J. A. King, R. B. Hester, and S. Ofori-Acquah
Activated neutrophil-mediated sickle red blood cell adhesion to lung vascular endothelium: role of phosphatidylserine-exposed sickle red blood cells
Am J Physiol Heart Circ Physiol,
October 1, 2006;
291(4):
H1679 - H1685.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Jacobi, S. Sela, H. I. Cohen, J. Chezar, and B. Kristal
Priming of polymorphonuclear leukocytes: a culprit in the initiation of endothelial cell injury
Am J Physiol Heart Circ Physiol,
May 1, 2006;
290(5):
H2051 - H2058.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. A. Neidlinger, S. K. Larkin, A. Bhagat, G. P. Victorino, and F. A. Kuypers
Hydrolysis of Phosphatidylserine-exposing Red Blood Cells by Secretory Phospholipase A2 Generates Lysophosphatidic Acid and Results in Vascular Dysfunction
J. Biol. Chem.,
January 13, 2006;
281(2):
775 - 781.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Mandal, A. Mazumder, P. Das, M. Kundu, and J. Basu
Fas-, Caspase 8-, and Caspase 3-dependent Signaling Regulates the Activity of the Aminophospholipid Translocase and Phosphatidylserine Externalization in Human Erythrocytes
J. Biol. Chem.,
November 25, 2005;
280(47):
39460 - 39467.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. J. Wu, L. Krishnamurti, J. L. Kutok, M. Biernacki, S. Rogers, W. Zhang, J. H. Antin, and J. Ritz
Evidence for ineffective erythropoiesis in severe sickle cell disease
Blood,
November 15, 2005;
106(10):
3639 - 3645.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. C. McMillan, C. L. Powell, Z. S. Bowman, J. D. Morrow, and D. J. Jollow
Lipids versus Proteins as Major Targets of Pro-Oxidant, Direct-Acting Hemolytic Agents
Toxicol. Sci.,
November 1, 2005;
88(1):
274 - 283.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. A. Coil and A. D. Miller
Enhancement of Enveloped Virus Entry by Phosphatidylserine
J. Virol.,
September 1, 2005;
79(17):
11496 - 11500.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. S. Bowman, J. D. Morrow, D. J. Jollow, and D. C. McMillan
Primaquine-Induced Hemolytic Anemia: Role of Membrane Lipid Peroxidation and Cytoskeletal Protein Alterations in the Hemotoxicity of 5-Hydroxyprimaquine
J. Pharmacol. Exp. Ther.,
August 1, 2005;
314(2):
838 - 845.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Sela, R. Shurtz-Swirski, M. Cohen-Mazor, R. Mazor, J. Chezar, G. Shapiro, K. Hassan, G. Shkolnik, R. Geron, and B. Kristal
Primed Peripheral Polymorphonuclear Leukocyte: A Culprit Underlying Chronic Low-Grade Inflammation and Systemic Oxidative Stress in Chronic Kidney Disease
J. Am. Soc. Nephrol.,
August 1, 2005;
16(8):
2431 - 2438.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. A. Coil and A. D. Miller
Phosphatidylserine Is Not the Cell Surface Receptor for Vesicular Stomatitis Virus
J. Virol.,
October 15, 2004;
78(20):
10920 - 10926.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. Shrestha, D. Gottlieb, and M. S. Diamond
Infection and Injury of Neurons by West Nile Encephalitis Virus
J. Virol.,
December 15, 2003;
77(24):
13203 - 13213.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Dias-Baruffi, H. Zhu, M. Cho, S. Karmakar, R. P. McEver, and R. D. Cummings
Dimeric Galectin-1 Induces Surface Exposure of Phosphatidylserine and Phagocytic Recognition of Leukocytes without Inducing Apoptosis
J. Biol. Chem.,
October 17, 2003;
278(42):
41282 - 41293.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Koshkaryev, S. Yedgar, H. Relevy, E. Fibach, and G. Barshtein
Acridine orange induces translocation of phosphatidylserine to red blood cell surface
Am J Physiol Cell Physiol,
September 1, 2003;
285(3):
C720 - C722.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Yasin, S. Witting, M. B. Palascak, C. H. Joiner, D. L. Rucknagel, and R. S. Franco
Phosphatidylserine externalization in sickle red blood cells: associations with cell age, density, and hemoglobin F
Blood,
July 1, 2003;
102(1):
365 - 370.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. G. Gallagher, S. H. Chang, M. P. Rettig, J. E. Neely, C. A. Hillery, B. D. Smith, and P. S. Low
Altered erythrocyte endothelial adherence and membrane phospholipid asymmetry in hereditary hydrocytosis
Blood,
June 1, 2003;
101(11):
4625 - 4627.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Ayi, G. Giribaldi, A. Skorokhod, E. Schwarzer, P. T. Prendergast, and P. Arese
16{alpha}-Bromoepiandrosterone, an Antimalarial Analogue of the Hormone Dehydroepiandrosterone, Enhances Phagocytosis of Ring Stage Parasitized Erythrocytes: a Novel Mechanism for Antimalarial Activity
Antimicrob. Agents Chemother.,
October 1, 2002;
46(10):
3180 - 3184.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Valles, M. T. Santos, J. Aznar, M. Martinez, A. Moscardo, M. Pinon, M. J. Broekman, and A. J. Marcus
Platelet-erythrocyte interactions enhance alpha IIbbeta 3 integrin receptor activation and P-selectin expression during platelet recruitment: down-regulation by aspirin ex vivo
Blood,
May 13, 2002;
99(11):
3978 - 3984.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Walsh, R. J. Lutz, T. G. Cotter, and R. O'Connor
Erythrocyte survival is promoted by plasma and suppressed by a Bak-derived BH3 peptide that interacts with membrane-associated Bcl-XL
Blood,
May 1, 2002;
99(9):
3439 - 3448.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. V. Tyulina, V. D. Prokopieva, R. D. Dodd, J. R. Hawkins, S. W. Clay, D. O. Wilson, A. A. Boldyrev, and P. Johnson
IN VITRO EFFECTS OF ETHANOL, ACETALDEHYDE AND FATTY ACID ETHYL ESTERS ON HUMAN ERYTHROCYTES
Alcohol Alcohol.,
March 1, 2002;
37(2):
179 - 186.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. N. Y. Setty, S. Kulkarni, and M. J. Stuart
Role of erythrocyte phosphatidylserine in sickle red cell-endothelial adhesion
Blood,
March 1, 2002;
99(5):
1564 - 1571.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. S. Kean, M. M. Durham, A. B. Adams, L. L. Hsu, J. R. Perry, D. Dillehay, T. C. Pearson, E. K. Waller, C. P. Larsen, and D. R. Archer
A cure for murine sickle cell disease through stable mixed chimerism and tolerance induction after nonmyeloablative conditioning and major histocompatibility complex-mismatched bone marrow transplantation
Blood,
March 1, 2002;
99(5):
1840 - 1849.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Eldor and E. A. Rachmilewitz
The hypercoagulable state in thalassemia
Blood,
January 1, 2002;
99(1):
36 - 43.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. N. Y. Setty, A. K. Rao, and M. J. Stuart
Thrombophilia in sickle cell disease: the red cell connection
Blood,
December 1, 2001;
98(12):
3228 - 3233.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. T. Wahid, S. M. Marshall, and T. H. Thomas
Increased Platelet and Erythrocyte External Cell Membrane Phosphatidylserine in Type 1 Diabetes and Microalbuminuria
Diabetes Care,
November 1, 2001;
24(11):
2001 - 2003.
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. de Jong, R. K. Emerson, J. Butler, J. Bastacky, N. Mohandas, and F. A. Kuypers
Short survival of phosphatidylserine-exposing red blood cells in murine sickle cell anemia
Blood,
September 1, 2001;
98(5):
1577 - 1584.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. de Jong, S. K. Larkin, L. A. Styles, R. M. Bookchin, and F. A. Kuypers
Characterization of the phosphatidylserine-exposing subpopulation of sickle cells
Blood,
August 1, 2001;
98(3):
860 - 867.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. N. Y. Setty, S. Kulkarni, C. D. Dampier, and M. J. Stuart
Fetal hemoglobin in sickle cell anemia: relationship to erythrocyte adhesion markers and adhesion
Blood,
May 1, 2001;
97(9):
2568 - 2573.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
B. N. Y. Setty, S. Kulkarni, A. K. Rao, and M. J. Stuart
Fetal hemoglobin in sickle cell disease: relationship to erythrocyte phosphatidylserine exposure and coagulation activation
Blood,
August 1, 2000;
96(3):
1119 - 1124.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. B. Manodori, G. A. Barabino, B. H. Lubin, and F. A. Kuypers
Adherence of phosphatidylserine-exposing erythrocytes to endothelial matrix thrombospondin
Blood,
February 15, 2000;
95(4):
1293 - 1300.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. BONOMINI, V. SIROLLI, N. SETTEFRATI, S. DOTTORI, L. DI LIBERATO, and A. ARDUINI
Increased Erythrocyte Phosphatidylserine Exposure in Chronic Renal Failure
J. Am. Soc. Nephrol.,
September 1, 1999;
10(9):
1982 - 1990.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
K. de Jong, S. K. Larkin, S. Eber, P. F.H. Franck, B. Roelofsen, and F. A. Kuypers
Hereditary Spherocytosis and Elliptocytosis Erythrocytes Show a Normal Transbilayer Phospholipid Distribution
Blood,
July 1, 1999;
94(1):
319 - 325.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Zhu, C. Paszty, T. Turetsky, S. Tsai, F. A. Kuypers, G. Lee, P. Cooper, P. G. Gallagher, M. E. Stevens, E. Rubin, et al.
Stomatocytosis Is Absent in "Stomatin"-Deficient Murine Red Blood Cells
Blood,
April 1, 1999;
93(7):
2404 - 2410.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K Muller, T Pomorski, P Muller, and A Herrmann
Stability of transbilayer phospholipid asymmetry in viable ram sperm cells after cryotreatment
J. Cell Sci.,
January 1, 1999;
112(1):
11 - 20.
[Abstract]
[PDF]
|
|
|
|
|
|
|
H. Hassoun, Y. Wang, J. Vassiliadis, M. Lutchman, J. Palek, L. Aish, I. S. Aish, S.-C. Liu, and A. H. Chishti
Targeted Inactivation of Murine Band 3 (AE1) Gene Produces a Hypercoagulable State Causing Widespread Thrombosis In Vivo
Blood,
September 1, 1998;
92(5):
1785 - 1792.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. R. Narasimhan, L. Yang, B. I. Gerwin, and V. C. Broaddus
Resistance of pleural mesothelioma cell lines to apoptosis: relation to expression of Bcl-2 and Bax
Am J Physiol Lung Cell Mol Physiol,
July 1, 1998;
275(1):
L165 - L171.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. A. Kuypers, J. Yuan, R. A. Lewis, L. M. Snyder, C. R. Kiefer, A. Bunyaratvej, S. Fucharoen, L. Ma, L. Styles, K. de Jong, et al.
Membrane Phospholipid Asymmetry in Human Thalassemia
Blood,
April 15, 1998;
91(8):
3044 - 3051.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. E. Boas, L. Forman, and E. Beutler
Phosphatidylserine exposure and red cell viability in red cell aging and in hemolytic anemia
PNAS,
March 17, 1998;
95(6):
3077 - 3081.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Bevers, P. Comfurius, D. Dekkers, M. Harmsma, and R. Zwaal
Review : Regulatory mechanisms of transmembrane phospholipid distributions and pathophysiological implications of transbilayer lipid scrambling
Lupus,
January 1, 1998;
7(2_suppl):
S126 - S131.
[Abstract]
[PDF]
|
|
|
|
|
|
|
R. F.A. Zwaal and A. J. Schroit
Pathophysiologic Implications of Membrane Phospholipid Asymmetry in Blood Cells
Blood,
February 15, 1997;
89(4):
1121 - 1132.
[Full Text]
[PDF]
|
|
|
|
|
