Role of hemichrome binding to erythrocyte membrane in the generation of band-3 alterations in beta-thalassemia intermedia erythrocytes

Mannu, Franca; Arese, Paolo; Cappellini, Maria Domenica; Fiorelli, Gernino; Cappadoro, Marina; Giribaldi, Giuliana; Turrini, Franco

doi:10.1182/blood.v86.5.2014.bloodjournal8652014

Cited by 94 publications

(68 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The clear supernatant (polydocanol extract) was immediately separated from the pellet and used for the gel-filtration chromatography. 22,23 Gel-Filtration Chromatography of Polydocanol Extracts. A 40-ϫ 1.0-cm column was filled with Sepharose CL-6B (excluded volume Ͼ4,000 kd molecular mass) and equilibrated with a solution containing (in mmol/L) 50 NaCl, 10 HEPES, 0.1% polydocanol, and pH 7.4.…”

Section: Preparation and Extraction Of Hypotonic Membranes With Noniomentioning

confidence: 99%

“…Ten micrograms membrane proteins was separated on 5% to 15% polyacrylamide gel and transferred to nitrocellulose as described. 22,23 After blocking for 1 hour in PBS containing 5% BSA, first antibodies were added to PBS containing 1% BSA. Anti-band 3 (cytoplasmic domain) antibody was diluted 1:1,000 in PBS and incubated for 1 hour at room temperature.…”

Section: Electrophoretic Separation Immunoblotting Of Membrane Protementioning

confidence: 99%

“…21 Therefore, we hypothesized that RBV may be responsible for membrane oxidative damage, which promotes a premature extravascular red cell removal similar to what was observed in congenital erythrocyte diseases, such as thalassemic syndromes, sickle cell disease, and G6PD deficiency. [22][23][24] We tested this hypothesis in vitro by evaluating indicators of erythrocyte oxidative susceptibility, such as hexosemonophosphate shunt (HMS) and in vivo by studying red cell features of a group of HCV patients undergoing RBV or RBV ϩ IFN administration. As observed in thalassemic erythrocyte, several indicators of erythrocyte oxidative susceptibility are affected by cell age; thus, in vivo we evaluated the band 3 aggregation state, which is primarily influenced by cell membrane oxidative damage and which mediates the red cell removal by reticuloendothelial system.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Hemolytic anemia induced by ribavirin therapy in patients with chronic hepatitis C virus infection: Role of membrane oxidative damage

et al. 2000

View full text Add to dashboard Cite

The antiviral drug ribavirin (RBV) is widely used in combination with interferon (IFN) in the treatment of chronic hepatitis C virus (HCV) infection. A major side effect of RBV is a reversible hemolytic anemia. We have evaluated the in vitro effects of RBV on erythrocyte adenosine triphosphate (ATP) content and on hexosemonophosphate shunt (HMS). The ATP levels were significantly decreased in the presence of RBV and the HMS was increased, suggesting the presence of red cell susceptibility to oxidation. In vivo, we have studied the hematologic effects of treatment with RBV alone or in combination with IFN in 11 patients with chronic hepatitis C: 6 were treated with RBV (1,000-1,200 mg/d) and 5 were treated with a combination of RBV and IFN (5 million U thrice weekly). Patients were studied at semi-monthly intervals from 0 to day 60 of therapy. Both treatments were associated with a significant reduction in hemoglobin levels (steady state level at day 45) and a marked increase in absolute reticulocyte counts. Erythrocyte Na-K pump activity was significantly diminished, whereas K-Cl cotransport and its dithiotreitol-sensitive fraction, malondialdehyde and methemoglobin levels were significantly increased. RBV-treated patients showed an increase in aggregated band 3, which was associated with a significantly increased binding of autologous antibodies and complement C3 fragments indicating an erithrophagocytic removal by reticuloendothelial system. (HEPATOLOGY 2000;31:997-1004.)Hepatitis C virus (HCV) infection is a major cause of chronic liver disease, leading to cirrhosis, end-stage liver disease, and hepatocellular carcinoma worldwide. 1,2 A major therapeutic goal in HCV-infected patients is to achieve early eradication of the virus, and to prevent severe long-term clinical complications. Interferon alfa (IFN-␣) is currently the only therapy that has been shown to have beneficial effects in chronic hepatitis type C. However, with a standard regimen of 3 million U administered 3 times per week for 6 to 12 months, only a small fraction of approximately 15% to 20% of the patients showed a sustained response with normalization of serum alanine transferase levels and serum HCV-RNA clearance. 3 Ribavirin (1-␤-D-ribofuranosyl-1H-1, 2,4-triazole-3-carboxamide) (RBV) is a water soluble synthetic guanosine analog that exerts antiviral activity against DNA and RNA viruses after intracellular phosphorylation. 4 Current studies indicate that combination therapy with RBV and IFN is associated with higher rates of sustained virological, biochemical, and histological response compared to IFN monotherapy. [5][6][7][8][9][10] The major side effect of RBV treatment is the occurrence of a reversible hemolytic anemia in a substantial proportion of treated patients. 11 The underlying mechanism is unknown. Studies on steady-state pharmacokinetics of RBV have shown that erythrocyte concentration of RBV greatly exceeds plasma concentrations 12 and that RBV is a transported permeant for the (es) nucleoside transporter in human erythrocytes...

show abstract

Section: Preparation and Extraction Of Hypotonic Membranes With Noniomentioning

confidence: 99%

Section: Electrophoretic Separation Immunoblotting Of Membrane Protementioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Hemolytic anemia induced by ribavirin therapy in patients with chronic hepatitis C virus infection: Role of membrane oxidative damage

et al. 2000

View full text Add to dashboard Cite

show abstract

“…Sodiumdodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was accomplished according to [17] with modifications of [18][19][20] and the evaluation of [21].…”

Section: Gel Electrophoresismentioning

confidence: 99%

Isolated erythrocyte membranes of transfusion‐dependent and non‐transfused thalassemia patients in Jakarta, investigated by electron paramagnetic resonance spectroscopy

Udyaningsih-Freisleben

Kurniati²,

Prasetyo

et al. 2003

BioFactors

View full text Add to dashboard Cite

Erythrocyte membrane structural parameters were studied in transfusion-dependent beta-thalassemia patients, in long-term transfused patients (regularly transfused < 15 years), and in those who had not yet obtained transfusions. Controls were voluntary students up to 30 years of age without diagnosis or clinical signs of thalassemia. Membranes were isolated and investigated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electron paramagnetic resonance (EPR) spectroscopy. Data obtained from the thiol-reactive spin label N-ethyl-maleimidoproxyl reveal immobilization of protein environment in erythrocyte membranes from thalassemic patients. SDS-PAGE shows both degradation and aggregation of membrane proteins. Thalassemic erythrocyte membranes exert higher order parameters in the hydrophobic region as determined by 16-doxyl-stearic acid. Rotational correlation times of this spin label increase only in transfused patients. Polarity is higher in membranes of all patients than in controls. In the polar interface, order parameters obtained from 5-doxyl-stearic acid increase in non-transfused and decrease in transfusion-dependent patients as compared with controls. Transfused patients exert increasing membrane order in the hydrophobic region and counter-currently decreasing order in the polar interface indicating loss of membrane integrity along with the loss of fluidity and polarity gradients and the loss the energetic barrier function of the membrane.

show abstract

“…We demonstrated IgG on erythroblasts and erythrocytes in a-and b-thalassaemia syndromes (Wiener et al, 1986(Wiener et al, , 1991Chinprasertsuk et al, 1997;Malasit et al, 1997) and provided evidence that it plays a role in the phagocytosis of b-thalassaemic erythrocytes by macrophages (MJ) (Wiener & Wickramasinghe, 1985). The IgG found on the surface of erythroid cells in thalassaemia is bound to clusters of band 3 protein (Yuan et al, 1992;Mannu et al, 1995;Cappellini et al, 1999) which has been aggregated due to the oxidant effect of membrane-1993; Wiener et al, 1996). In addition, in b-thal/HbE we found increased expression of class I Fcg receptor (FcgRI) (CD64) on Mo and granulocytes (Wanachiwanawin et al, 1993).…”

mentioning

confidence: 68%

Role of FcγRI (CD64) in erythrocyte elimination and its up‐regulation in thalassaemia

Wiener¹,

Allen²,

Porter³

et al. 1999

Br J Haematol

View full text Add to dashboard Cite

To examine any role of the high affinity Fcgamma class I receptor (FcgammaRI) (CD64) in erythrocyte elimination by mononuclear phagocytes (MP) in thalassaemia (thal), we investigated the in vitro interaction of beta-thalassaemic erythrocytes with monocytes (Mo) whose FcgammaR expression had been modulated by cytokines. Treatment of Mo with interferon (IFN)-gamma or interleukin (IL)-10 which up-regulate FcgammaRI, caused a dose-dependent increase in binding of beta-thalassaemic erythrocytes, whereas stimulation with IL-4 which down-regulates the receptor, reduced this interaction, in a dose-dependent manner, to that of normal erythrocytes. Binding of thalassaemic erythrocytes by IFN-gamma or IL-10-treated Mo was inhibited by FcgammaRI-specific reagents. In addition, Mo expression of FcgammaRI and HLA class II DR was determined by flow cytometry in Thai patients with HbH disease (alpha1/alpha2 or alpha1/Hb Constant Spring) (n = 15) or beta degrees -thal/HbE (n = 16). In both groups of patients FcgammaRI expression was increased as compared to normal controls (n = 14): mean fluorescence intensity (+/-SD) 124.79 +/- 38. 77 in HbH disease and 121.86 +/- 18.23 in beta degrees -thal/HbE versus 91.94 +/- 17.36 in normal controls (P < 0.01 and P < 0.001, respectively). In contrast, HLA class II DR expression was similar in patients and controls. The results suggest that, in thalassaemia, up-regulated FcgammaRI on mononuclear phagocytes plays a role in their interaction with erythrocytes and that this process can be modified by cytokines.

show abstract

Role of hemichrome binding to erythrocyte membrane in the generation of band-3 alterations in beta-thalassemia intermedia erythrocytes

Cited by 94 publications

References 31 publications

Hemolytic anemia induced by ribavirin therapy in patients with chronic hepatitis C virus infection: Role of membrane oxidative damage

Hemolytic anemia induced by ribavirin therapy in patients with chronic hepatitis C virus infection: Role of membrane oxidative damage

Isolated erythrocyte membranes of transfusion‐dependent and non‐transfused thalassemia patients in Jakarta, investigated by electron paramagnetic resonance spectroscopy

Role of FcγRI (CD64) in erythrocyte elimination and its up‐regulation in thalassaemia

Contact Info

Product

Resources

About