Abstract
Bone marrow transplantation (BMT) serves as the only curative treatment for patients with β-thalassemia major; however, hemostatic changes have been observed in these BMT patients. Aggregability of thalassemic red blood cells (RBCs) and increased red blood cell-derived microparticles (RMPs) expressing phosphatidylserine (PS) are thought to participate in thromboembolic events by initially triggering platelet activation. To our knowledge, there has been no report providing quantitation of these circulating PS-expressing RBCs and RMPs in young β-thalassemia patients after BMT. Whole blood from each subject was fluorescently labeled to detect RBC markers (CD235a) and annexin-V together with the known number TruCount™ beads. PS-expressing RBCs, RMPs, and activated platelets were identified by flow cytometry. In our randomized study, we found the decreased levels of three aforementioned factors compared to levels in patients receiving regular blood transfusion (RT). This study showed that BMT in β-thalassemia patients decreases the levels of circulating PS-expressing RBCs, their MPs, and procoagulant platelets when compared to patients who received RT. Normalized levels of these coagulation markers may provide the supportive evidence of the effectiveness of BMT for curing thalassemia.
Similar content being viewed by others
References
Fucharoen S, Winichagoon P (2002) Thalassemia and abnormal hemoglobin. Int J Hematol 76(Suppl 2):83–89
Fucharoen S, Winichagoon P (2011) Haemoglobinopathies in southeast Asia. Indian J Med Res 134:498–506
Hongeng S, Pakakasama S, Chaisiripoomkere W, Chuansumrit A, Sirachainan N, Ungkanont A et al (2004) Outcome of transplantation with unrelated donor bone marrow in children with severe thalassaemia. Bone Marrow Transplant 33(4):377–379
Anurathapan U, Hongeng S, Pakakasama S, Sirachainan N, Songdej D, Chuansumrit A et al (2016) Hematopoietic stem cell transplantation for homozygous beta-thalassemia and beta-thalassemia/hemoglobin E patients from haploidentical donors. Bone Marrow Transplant 51(6):813–818
Issaragrisil S, Kunacheewa C (2016) Matched sibling donor hematopoietic stem cell transplantation for thalassemia. Curr Opin Hematol 23(6):508–514
Rund D, Rachmilewitz E (2005) Beta-thalassemia. N Engl J Med 353(11):1135–1146
Freikman I, Amer J, Cohen JS, Ringel I, Fibach E (2008) Oxidative stress causes membrane phospholipid rearrangement and shedding from RBC membranes—an NMR study. Biochim Biophys Acta 1778(10):2388–2394
Kuypers FA, Yuan J, Lewis RA, Snyder LM, Kiefer CR, Bunyaratvej A et al (1998) Membrane phospholipid asymmetry in human thalassemia. Blood 91(8):3044–3051
Eldor A, Rachmilewitz EA (2002) The hypercoagulable state in thalassemia. Blood 99(1):36–43
Mankelow TJ, Griffiths RE, Trompeter S, Flatt JF, Cogan NM, Massey EJ et al (2015) Autophagic vesicles on mature human reticulocytes explain phosphatidylserine-positive red cells in sickle cell disease. Blood 126(15):1831–1834
de Jong K, Larkin SK, Styles LA, Bookchin RM, Kuypers FA (2001) Characterization of the phosphatidylserine-exposing subpopulation of sickle cells. Blood 98(3):860–867
Basu S, Banerjee D, Ghosh M, Chakrabarti A (2010) Erythrocyte membrane defects and asymmetry in paroxysmal nocturnal hemoglobinuria and myelodysplastic syndrome. Hematology 15(4):236–239
Zahedpanah M, Azarkeivan A, Aghaieepour M, Nikogoftar M, Ahmadinegad M, Hajibeigi B et al (2014) Erythrocytic phosphatidylserine exposure and hemostatic alterations in beta-thalassemia intermediate patients. Hematology 19(8):472–476
Pattanapanyasat K, Noulsri E, Fucharoen S, Lerdwana S, Lamchiagdhase P, Siritanaratkul N et al (2004) Flow cytometric quantitation of red blood cell vesicles in thalassemia. Cytometry B Clin Cytom 57(1):23–31
Xiong Z, Oriss TB, Cavaretta JP, Rosengart MR, Lee JS (2012) Red cell microparticle enumeration: validation of a flow cytometric approach. Vox Sang 103(1):42–48
Rubin O, Crettaz D, Canellini G, Tissot JD, Lion N (2008) Microparticles in stored red blood cells: an approach using flow cytometry and proteomic tools. Vox Sang 95(4):288–297
Pattanapanyasat K, Gonwong S, Chaichompoo P, Noulsri E, Lerdwana S, Sukapirom K et al (2007) Activated platelet-derived microparticles in thalassaemia. Br J Haematol 136(3):462–471
Lamchiagdhase P, Nitipongwanich R, Rattanapong C, Noulsri E, Lerdwana S, Pattanapanyasat K (2004) Red blood cell vesicles in thalassemia. J Med Assoc Thail 87(3):233–238
Thomas ED, Buckner CD, Sanders JE, Papayannopoulou T, Borgna-Pignatti C, De Stefano P et al (1982) Marrow transplantation for thalassaemia. Lancet 2(8292):227–229
Sabloff M, Chandy M, Wang Z, Logan BR, Ghavamzadeh A, Li CK et al (2011) HLA-matched sibling bone marrow transplantation for beta-thalassemia major. Blood 117(5):1745–1750
Boas FE, Forman L, Beutler E (1998) Phosphatidylserine exposure and red cell viability in red cell aging and in hemolytic anemia. Proc Natl Acad Sci U S A 95(6):3077–3081
Kuypers FA, de Jong K (2004) The role of phosphatidylserine in recognition and removal of erythrocytes. Cell Mol Biol (Noisy-le-Grand, France) 50(o):147–158
Basu S, Banerjee D, Chandra S, Chakrabarti A (2008) Loss of phospholipid membrane asymmetry and sialylated glycoconjugates from erythrocyte surface in haemoglobin E beta-thalassaemia. Br J Haematol 141(1):92–99
Sirachainan N, Thongsad J, Pakakasama S, Hongeng S, Chuansumrit A, Kadegasem P et al (2012) Normalized coagulation markers and anticoagulation proteins in children with severe beta-thalassemia disease after stem cell transplantation. Thromb Res 129(6):765–770
Tantawy AA, Adly AA, Ismail EA, Habeeb NM (2013) Flow cytometric assessment of circulating platelet and erythrocytes microparticles in young thalassemia major patients: relation to pulmonary hypertension and aortic wall stiffness. Eur J Haematol 90(6):508–518
Schrier SL (2002) Pathophysiology of thalassemia. Curr Opin Hematol 9(2):123–126
Voskou S, Aslan M, Fanis P, Phylactides M, Kleanthous M (2015) Oxidative stress in beta-thalassaemia and sickle cell disease. Redox Biol 6:226–239
Jevtic D, Vujic D, Zecevic Z, Veljkovic D, Gazikalovic S, Elezovic I (2010) Coagulation disturbances in paediatric patients with hepatic veno-occlusive disease after stem cells transplantation. Srp Arh Celok Lek 138(Suppl 1):33–38
Klaihmon P, Phongpao K, Kheansaard W, Noulsri E, Khuhapinant A, Fucharoen S et al (2017) Microparticles from splenectomized beta-thalassemia/HbE patients play roles on procoagulant activities with thrombotic potential. Ann Hematol 96(2):189–198
Rank A, Nieuwland R, Toth B, Pihusch V, Delker R, Hiller E et al (2011) Microparticles for diagnosis of graft-versus-host disease after allogeneic stem transplantation. Transplantation 92(2):244–250
Trummer A, De Rop C, Stadler M, Ganser A, Buchholz S (2011) P-selectin glycoprotein ligand-1 positive microparticles in allogeneic stem cell transplantation of hematologic malignancies. Exp Hematol 39(11):1047–1055
Acknowledgements
We thank the staff at the Hematology-Oncology Division, Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, for recruiting subjects and helping in blood sample collection. Thanks also go to Dr. Arthur E. Brown for editing the manuscript. PK and SV were financially supported by the Thailand Research Fund (TRF) RGJ-PhD scholarship. EN thanks the Faculty of Medicine Siriraj Hospital, Mahidol University, for supporting the research project. KP was the recipient of the TRF-Distinguished Research Professor Award (grant contact number DPG5980001).
Author information
Authors and Affiliations
Contributions
PK, SV, EN performed the experiments, analyzed the data and wrote manuscript; SL, UA, NS supervised the project and analyzed the data; SH, KP designed the experiment, wrote, and edited the paper.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Electronic supplementary material
Supplementary Table 1
Demographic data of two patient groups. (DOCX 17 kb)
Rights and permissions
About this article
Cite this article
Klaihmon, P., Vimonpatranon, S., Noulsri, E. et al. Normalized levels of red blood cells expressing phosphatidylserine, their microparticles, and activated platelets in young patients with β-thalassemia following bone marrow transplantation. Ann Hematol 96, 1741–1747 (2017). https://doi.org/10.1007/s00277-017-3070-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00277-017-3070-2