Erythrocyte membrane skeleton abnormalities in severe beta-thalassemia

Shinar, Eilat; Shalev, Oded; Ea, Rachmilewitz; Schrier, SL

doi:10.1182/blood.v70.1.158.bloodjournal701158

Cited by 31 publications

(22 citation statements)

References 33 publications

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“…These include extraordinary ineffective erythropoiesis in the marrow (6) and altered deformability due to the rigidity of the RBC membrane, which impairs passage of RBCs through sinusoidal walls of reticuloendothelial organs, and which finally triggers the removal of these cells from circulation (2–5, 7). In addition, the rigidity of RBC membrane is associated with increased aggregability of RBCs, as has been documented in both sickle‐cell anemia and thalassemia (8, 9). RBC aggregability is an important determinant of blood flow, particularly in the microcirculation (10).…”

Section: Introductionmentioning

confidence: 84%

Flow cytometric quantitation of red blood cell vesicles in thalassemia

Pattanapanyasat

Noulsri

Fucharoen

et al. 2003

Cytometry Part B Clinical

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Background: Thalassemia is a hereditary hemolytic anemia caused by mutations in the globin gene complex. Circulatory disturbances including arterial and venous thrombosis have also been noted in these patients. Aggregability of abnormal RBC and the high level of membrane-derived microparticles stemming from activated platelets and other blood cells are thought to be responsible for the associated thrombotic risk. Destruction of RBC is also thought to be an important pathophysiological consequence, particularly through the formation of circulating vesicles. To our knowledge, there has been no attempt to quantitatively evaluate the number of RBC vesicles in thalassemia. This prompted us to study the level of RBC vesicles in the peripheral blood of thalassemia patients using quantitative flow cytometry.Methods: Whole blood from each subject was doubly stained for RBC and platelet or annexin V markers, together with the known density TruCount™ beads. RBC vesicles were gated according to their forward/side scatter and RBC marker. Percentage of RBC vesicles and their absolute number were analyzed by flow cytometry.Results: Our data indicated that RBC vesicles were annexin V-positive. The number of annexin V-positive events was higher than their intact RBCs. RBC vesicles were present in both normal and thalassemic blood samples, but the numbers of RBC vesicles were significantly higher in thalassemia. Both the percentage and the absolute number of RBC vesicles were especially marked in splenectomized subjects with ␤-thalassemia/ Hemoglobin E. When clinical and hematological indices were compared with RBC vesicles, there was an inverse relationship between the degree of severity in thalassemia patients and the number of RBC vesicles.Conclusion: Flow cytometric quantitation of RBC vesicles is simple, reliable and may offer new insights in to study of the relationship between defective hemoglobin synthesis, RBC perturbation and pathophysiological complications in thalassemia.

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Section: Introductionmentioning

confidence: 84%

Flow cytometric quantitation of red blood cell vesicles in thalassemia

Pattanapanyasat

Noulsri

Fucharoen

et al. 2003

Cytometry Part B Clinical

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“…For example, the RBC membrane undergoes vesiculation under a variety of conditions, which include increased cytoplasmic calcium concentration, reduced ATP content and disruption of the membrane lipid-protein organization (Wagner et al, 1986)all of which are features characteristic of thalassaemic RBCs. It has been shown that vesicles and MPs from platelets and RBC strongly bind annexin V, a protein known for its interaction with negatively charged phospholipids such as phosphatidylserine (PS) (Shinar et al, 1987;Borenstain-Ben Yashar et al, 1993;Zwaal & Schroit, 1997;Kuypers et al, 1998). PS is one of the key membrane phospholipids that is normally located along the inner side of the membrane bilayer.…”

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confidence: 99%

Activated platelet‐derived microparticles in thalassaemia

et al. 2006

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Summary Thromboembolic complications have been documented in thalassaemia patients. The aggregability of abnormal red blood cells and the high level of membrane‐derived microparticles (MPs) stemming from blood cells are thought to be responsible for the associated thrombotic risk. We investigated the number of MPs, their cellular origin and their procoagulant properties in β‐thalassaemia. Fresh whole blood was simultaneously stained for annexin V, cellular antigens and the known density beads. The procoagulant properties of these phosphatidylserine (PS)‐bearing MPs were also measured by assessing the platelet factor‐3‐like activity in the blood. Flow cytometric results showed that splenectomised β‐thalassaemia/HbE patients had significantly higher levels of PS‐bearing MPs than non‐splenectomised β‐thalassaemia/HbE patients and normal individuals (P < 0·0001). There was a good correlation between PS‐bearing MPs and PS‐bearing platelets, reflecting the existence of chronic platelet activation in β‐thalassaemia/HbE patients (rs = 0·511, P < 0·001). The cellular origin of PS‐bearing MPs showed mostly activated‐platelet origin with adhesion (CD41a/CD62P/CD36). Moreover, the platelet procoagulant activity was higher in splenectomised β‐thalassaemia/HbE patients when compared with non‐splenectomised (P < 0·05) and normal individuals (P < 0·01), and the amount correlated with PS‐bearing MPs (rs = 0·560, P < 0·001). These findings suggest that MPs originate from activated platelets with a potential to aggravate thrombotic events when the numbers are excessive, as is commonly seen in splenectomised β‐thalassaemia/HbE patients.

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“…This can be confirmed by lower GSH concentration found in thalassaemic RBCs (Table 3). Oxidative stress, in fact, is known to cross-link integral membrane proteins as dimers and tetramers of band 3 (Salhany et al, 1990;Turrini et al, 1994;Giger et al, 1995;Adragna and Lauf, 1997;Datta et al, 2006), and to lead to a decrease in and oxidation of membrane thiols (Shinar et al, 1987;Olivieri et al, 1994). Table 4 Fe concentration (mg/ml) in normal and thalassaemic plasma and erythrocytes Data are presented as means¡S.D.…”

Section: Discussionmentioning

confidence: 99%

Erythrocytes anion transport and oxidative change in β‐thalassaemias

Maria

Romano²,

Romano

et al. 2010

Cell Biology International

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beta-Thalassaemia is characterized by a decrease in globin beta-chain synthesis and an excess in free alpha-globin chains. This induces alterations in membrane lipids and proteins resulting from a reduction in spectrin/band 3 ratio, partial oxidation of band 4.1 and clustering of band 3. The membrane injury provokes hyperhaemolysis and bone marrow hyperplasia. The pathophysiology of thalassaemia is associated with iron overload that generates oxygen free radicals and oxidative tissue injury with ocular vessel alterations. The aim of this research is to investigate the influence of oxidative stress on band 3 efficiency, which is an integral membrane protein of RBCs (red blood cells). Band 3 protein, of which there are more than 1 million copies per cell, is the most abundant membrane protein in human RBCs. It mediates the anion exchange and acid-base equilibrium through the RBC membrane. Some experiments were performed on thalassaemic cells and beta-thalassaemia-like cells and tested for sulfate uptake. To test the antioxidant effect of Mg(2+), other experiments were performed using normal and pathological cells in the presence of Mg(2+). The oxidant status in thalassaemic cells was verified by increased K(+) efflux, by lower GSH levels and by increased G6PDH (glucose-6-phosphate dehydrogenase) activity. The rate constant of SO(4)(2-) uptake decreases in thalassaemic cells as well as in beta-thalassaemia-like cells when compared with normal cells. It increases when both cells are incubated with Mg(2+). Our data show that oxidative stress plays a relevant role in band 3 function of thalassaemic cells and that antioxidant treatment with Mg(2+) could reduce oxidative damage to the RBC membrane and improve the anion transport efficiency regulated by band 3 protein.

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Erythrocyte membrane skeleton abnormalities in severe beta-thalassemia

Cited by 31 publications

References 33 publications

Flow cytometric quantitation of red blood cell vesicles in thalassemia

Flow cytometric quantitation of red blood cell vesicles in thalassemia

Activated platelet‐derived microparticles in thalassaemia

Erythrocytes anion transport and oxidative change in β‐thalassaemias

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