Erythropoietin Administration Increases Production and Reactivity of Platelets in Dogs

Summary

Administration of erythropoietin (EPO) to adult dogs resulted in a dramatic increase in the number of thiazole orange-positive (TO⁺) platelets, also referred to as reticulated platelets. Pre-treatment level of TO⁺ platelets was 6.2 ± 0.5% (mean ± 1 SE; n = 5); following day 5 of treatment with 500 U EPO/kg/day, the percentage of TO⁺ platelets peaked at 16.8 ± 2.3% (n = 5; p <0.02). After cessation of the hormone, the number of TO⁺ platelets fell rapidly to below starting levels. Unexpectedly, there was a significant decline in total platelet count during EPO administration despite an increased level of TO⁺ platelets. To assess platelet reactivity, total platelets and TO⁺ platelets from EPO- treated dogs were analyzed for thrombin-responsiveness as quantitated by P-selectin expression on the cell surface; reactivity was expressed as a thrombin EC₅₀, the thrombin concentration required to activate 50% of platelets. Both total and TO⁺ platelets were hyperreactive during EPO treatment when compared either to pre-treatment values or to control animals. Thrombin EC₅₀ values for total and TO⁺ platelets on day 5 fell to 66.5 ± 5.4% (mean ± 1 SE; n = 5; p <0.02) and 62.2 ± 8.7% (n = 5; p <0.025), respectively, of pre-treatment levels. These data indicate that EPO not only promotes the synthesis of increased numbers of TO⁺ platelets in the dog but that these newly produced platelets are hyperreactive when compared to TO⁺ platelets from control animals.

• References

• 1 Krantz SB. Erythropoietin. Blood 1991; 77: 419-434
• 2 Ishibashi T, Koziol JA, Burstein SA. Human recombinant erythropoietin promotes differentiation of murine megakaryocytes in vitro. J Clin Invest 1987; 79: 286-289
• 3 Papayannopoulou T, Brice M, Farrer D, Kaushansky K. Insights into the cellular mechanisms of erythropoietin-thrombopoietin synergy. Exp Hematol 1996; 24: 660-669
• 4 Fraser JK, Tan AS, Lin FK, Berridge MV. Expression of specific high-affinity binding sites for erythropoietin on rat and mouse megakaryocytes. Exp Hematol 1989; 17: 10-16
• 5 Berridge MV, Fraser JK, Carter JM, Lin FK. Effects of recombinant human erythropoietin on megakaryoctyes and on platelet production in the rat. Blood 1988; 72: 970-977
• 6 Tsukada J, Misago M, Kikuchi M, Sato T, Ogawa R, Oda S, Chiba S, Eto S. The effect of high doses of recombinant human erythropoietin on megakaryocytopoiesis and platelet production in splenectomized mice. Br J Haematol 1990; 76: 260-268
• 7 Shikama Y, Ishibashi T, Kimura H, Kawaguchi M, Uchida T, Maruyama Y. Transient effect of erythropoietin on thrombocytopoiesis in vivo in mice. Exp Hematol 1992; 20: 216-222
• 8 Pirisi M, Fabris C, Soardo G, Cecchin E, Toniutto P, Bartoli E. Thrombocytopenia of chronic liver disease corrected by erythropoietin treatment. J Hepatology 1994; 21: 376-380
• 9 Sharpe PC, Desai ZR, Morris TCM. Increase in mean platelet volume in patients with chronic renal failure treated with erythropoietin. J Clin Pathol 1994; 47: 159-161
• 10 Van Geet C, Hauglustaine D, Verresen L, Vanrusselt M, Vermylen J. Haemostatic effects of recombinant human erythropoietin in chronic haemodialysis patients. Thromb Haemost 1989; 61: 117-121
• 11 Malyszko J, Malyszko JS, Borawski J, Rydzewski A, Kalinowski M, Azzadin A, Mysliwiec M, Buczko W. A study of platelet functions some hemostatic and fibrinolytic parameters in relation to serotonin in hemodialyzed patients under erythropoietin therapy. Thromb Res 1995; 77: 133-143
• 12 Shattil SJ, Cunningham M, Hoxie JA. Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies. Blood 1987; 70: 307-315
• 13 Johnston GI, Pickett EB, McEver RP, George JN. Heterogeneity of platelet secretion in response to thrombin demonstrated by fluorescence flow cytometry. Blood 1987; 69: 401-403
• 14 Peng J, Friese P, Wolf RF, Harrison P, Downs T, Lok S, Dale GL, Burstein SA. Relative reactivity of platelets from thrombopoietin- and interleukin-6-treated dogs. Blood 1996; 87: 4158-4163
• 15 Dale GL, Friese P, Hynes LA, Burstein SA. Demonstration that thiazole orange-positive platelets in the dog are less than twenty-four hours old. Blood 1995; 85: 1822-1825
• 16 Peng J, Friese P, Heilmann E, George JN, Burstein SA, Dale GL. Aged platelets have an impaired response to thrombin as quantitated by P-selectin expression. Blood 1994; 83: 161-166
• 17 Burstein SA, Friese P, Downs T, Epstein RE. Canine megakaryocytopoiesis: Analysis utilizing a monoclonal antibody to a 140 kD dog platelet protein. Exp Hematol 1991; 19: 47-52
• 18 Ault KA, Rinder HM, Mitchell J, Carmody MB, Vary CPH, Hillman RS. The significance of platelets with increased RNA content (reticulated platelets). Am J Clin Pathol 1992; 98: 637-646
• 19 Lee LG, Chen CH, Chiu LA. Thiazole orange: A new dye for reticulocyte analysis. Cytometry 1986; 7: 508-517
• 20 Eschbach JW, Abudulhadi MH, Browne JK, Delano BG, Downing MR, Egrie JC, Evans RW, Friedman EA, Graber SE, Haley NR, Korbert S, Krantz SB, Lundin AP, Nissenson AR, Ogden DA, Paganini EP, Rader B, Rutsky EA, Stivelman J, Stone WJ, Teschan P, Van Stone JC, Van Wyck DB, Zuckerman K, Adamson JW. Recombinant human erythropoietin in anemic patients with end-stage renal disease. Ann Int Med 1989; 111: 0992-1000
• 21 Taylor JE, Belch JJ, Henderson IS, Stewart WK. Erythropoietin does not increase whole blood platelet aggregation in vitro. Nephrology, Dialysis, Transplantation 1993; 8: 556-558
• 22 McDonald TP, Clift RE, Cottrell MB. Large, chronic doses of erythropoietin cause thrombocytopenia in mice. Blood 1992; 80: 352-358
• 23 Wolf RF, Gilmore LS, Friese P, Downs T, Burstein SA, Dale GL. Erythropoietin potentiates thrombus development in a canine arterio-venous shunt model. Thromb Haemost 1997; 77: 1020-1024
• 24 Macdougall IC, Roberts DE, Neubert P, Dharmasena AD, Coles GA, Williams JD. Pharmacokinetics of intravenous intraperitoneal and subcutaneous recombinant erythropoietin in patients with CAPD. Contrib Nephrol 1989; 76: 112-121
• 25 Erslev AJ, Wilson J, Caro J. Erythropoietin titers in anemic nonuremic patients. J Lab Clin Med 1987; 109: 429-433
• 26 Balkaran B, Char G, Morris JS, Thomas PW, Serjeant BE, Serjeant GR. Stroke in a cohort of patients with homozygous sickle cell disease. J Pediatrics 1992; 120: 360-366
• 27 Palek J, Jarolim P. Hereditary spherocytosis elliptocytosis and related disorders. In: Williams Hematology Beutler E, Lichtman MA, Coller BS, Kipps TJ. (eds) 5. McGraw-Hill; New York: 1995. pp 536-557
• 28 Hirsh J, Dacie JV. Persistent post-splenectomy thrombocytosis and thromboembolism: A consequence of continuing anaemia. Br J Haematol 1966; 12: 44-53
• 29 Lipski DA, Bergamini TM. Upper extremity arterial thrombosis associated with immunohemolytic anemia. Surgery 1996; 119: 354-355
• 30 Brown AS, Martin JF. The megakaryocyte platelet system and vascular disease. Eur J Clin Invest 1994; (Suppl. 24) 1: 09-15
• 31 Harker LA, Hanson SR. Platelet factors predisposing to arterial thrombosis. Bailliere’s Clin Haematol 1994; 7: 499-522