Assay of an Activity in the Serum of Patients with Disorders of Thrombopoiesis That Stimulates Formation of Megakaryocytic Colonies

Hoffman, Ronald; Mazur, Eric Michael; Bruno, Edward; Floyd, Victoria

doi:10.1056/nejm198109033051001

Cited by 113 publications

(45 citation statements)

References 22 publications

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“…[21][22][23][24] On the other hand, it was described that TPO levels were high when thrombocytopenia was due to megakaryocyte deficiency, and low when due to increased platelet destruction, 19 and that serum megakaryocyte colony-stimulating activity levels appeared to be inversely related to marrow megakaryocyte numbers. 25 In the present study, we could clearly demonstrate using a sensitive ELISA that human TPO levels also changed inversely with the number of platelets after myelosuppressive chemotherapy. Furthermore, when platelets were transfused into patients at the time of nadir of platelet counts, elevated TPO levels dropped rapidly by around 4.0 fmol/ml within 1 h of the platelet transfusion, while platelet counts increased by around 28 × 10 9 /l.…”

Section: Discussionsupporting

confidence: 58%

Serum thrombopoietin levels in patients correlate inversely with platelet counts during chemotherapy-induced thrombocytopenia

et al. 1998

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We studied serum thrombopoietin (TPO) levels and circulating numbers of platelet during five courses of myelosuppressive post-remission chemotherapy in three patients with acute leukemia in complete remission. Serum TPO levels were measured by a newly developed and sensitive sandwich enzyme-linked immunosorbent assay. In all courses, serum TPO levels changed reciprocally with the platelet counts. When platelets were transfused into patients near the time of platelet nadir, the TPO levels dropped temporarily, while platelet counts temporarily increased. In addition, platelets obtained after transfusion in a thrombocytopenic patient showed lower binding to biotinylated TPO than donor platelets prior to the transfusion. The finding indicated that the TPO receptors were saturated with endogenous TPO of the patient with a high serum TPO level. These results suggest that the platelet mass directly regulates serum TPO levels by receptor-mediated absorption and is one of the major regulators of serum TPO levels in humans.

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Section: Discussionsupporting

confidence: 58%

Serum thrombopoietin levels in patients correlate inversely with platelet counts during chemotherapy-induced thrombocytopenia

et al. 1998

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“…16,17 Attempts to use megakaryocyte colony formation to study thrombopoiesis in ITP have given contradictory results. Some groups, culturing ITP bone marrow, have found increased numbers of megakaryocyte colony-forming units (CFU-Ms), 18,19 whereas others have found decreased numbers of CFU-Ms. 20 Similarly, one group 21 reported that ITP plasma did not affect CFU-M formation by normal cells, yet another group 18 noted increased CFU-M formation in the presence of ITP serum.…”

Section: Discussionmentioning

confidence: 99%

Suppression of in vitro megakaryocyte production by antiplatelet autoantibodies from adult patients with chronic ITP

McMillan

Wang²,

Tomer³

et al. 2004

Blood

460

319

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Chronic immune thrombocytopenic purpura (ITP) is manifested by autoantibodyinduced platelet destruction. Platelet turnover studies suggest that autoantibody may also affect platelet production. To evaluate this, we studied the effect of plasma from adult patients with chronic ITP on in vitro megakaryocyte production. CD34 ؉ cells, obtained from healthy donors, were cultured in medium containing PEG-rHuMGDF and 10% plasma from either ITP patients or healthy subjects. Cultures containing plasma from 12 of 18 ITP patients showed a significant decrease (26%-95%) in megakaryocyte production when compared with control cultures. Positive ITP plasmas not only reduced the total number of megakaryocytes produced during the culture period but also inhibited megakaryocyte maturation, resulting in fewer 4N, 8N, and 16N cells. The role of antibody in this suppression is supported by 2 factors: (1) immunoglobulin G (IgG) from ITP patients inhibited megakaryocyte production when compared with control IgG; and (2) adsorption of autoantibody, using immobilized antigen, resulted in significantly less inhibition of megakaryocyte production when compared with unadsorbed plasma. These results show that plasma autoantibody from some adult patients with ITP inhibits in vitro megakaryocyte production, suggesting that a similar effect may occur in vivo. IntroductionIn 1950, Dr William Harrington was infused with blood from a patient with chronic immune thrombocytopenic purpura (ITP), resulting in the rapid onset of severe thrombocytopenia. He recovered within the next few days. Further studies by Harrington et al 1 and Shulman et al 2 showed clearly that patients with chronic ITP have a circulating factor capable of destroying homologous and autologous platelets. The severity of the thrombocytopenia was dose dependent, and the plasma factor could be adsorbed by platelets and was present in the immunoglobulin G (IgG)-rich fraction after ion-exchange chromatography. For the next several years, it was concluded that thrombocytopenia in patients with chronic ITP was caused solely by autoantibody-induced platelet destruction.However, in the early 1980s, autologous platelet survival studies from several laboratories showed that in approximately two thirds of ITP patients, platelet turnover is either reduced or normal, not increased, as would be expected if platelet destruction were the only mechanism causing thrombocytpenia. [3][4][5][6] Because megakaryocytes express GPIIb-IIIa and GPIb-IX on their surfaces during maturation 7 and because most ITP autoantibodies react with one or both of these glycoprotein complexes, 8,9 it follows that autoantibody binding to megakaryocytes could interfere with platelet production and release from the bone marrow either by causing intramedullary megakaryocyte or platelet destruction or by interfering with megakaryocyte maturation.Recently, Chang et al 10 evaluated the effect of plasma, from patients with childhood ITP (44 with acute ITP and 9 with chronic ITP), on thrombopoietin-induced production of megaka...

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“…Urine (5,6), serum (7), and plasma (8,9) obtained from patients with hypomegakaryocytic thrombocytopenia have been shown to promote the formation of megakaryocyte colonies by marrow mononuclear cells in vitro. Phytohemagglutininstimulated leukobyte-conditioned media as well as normal bone and lung conditioned media contain a similar activity (10).…”

Section: Introductionmentioning

confidence: 99%

Purification and partial characterization of a megakaryocyte colony-stimulating factor from human plasma.

Hoffman¹,

Yang²,

Bruno³

et al. 1985

J. Clin. Invest.

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Human plasma obtained from patients with hypomegakaryocytic thrombocytopenia contains a factor that promotes megakaryocyte colony formation by normal human marrow cells. This megakaryocyte colony-stimulating factor was purified from such a plasma specimen. A four-step purification scheme which included ammonium sulfate precipitation, diethylaminoethylSepharose chromatography, affinity chromatography on wheat germ lectin-Sepharose 6MB, and reverse-phase high performance liquid chromatography resulted in a recovery of 16.6% of the initial biological activity and an increase in specific activity by 3,489-fold. The purified protein produced a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purified megakaryocyte colony-stimulating factor was capable of promoting megakaryocyte colony formation at a concentration of 7.6 X iO-M. Megakaryocyte colony-stimulating factor was shown to be a glycoprotein and had an apparent 46,000 mol wt. Deglycosylation of megakaryocyte colony-stimulating factor by treatment with trifluoromethanesulfonate resulted in the loss of its ability to promote megakaryocyte colony formation. Megakaryocyte colony-stimulating factor appears to be an important regulator of in vitro human megakaryocytopoiesis at the level of the colony-forming unit megakaryocyte and may be of importance physiologically.

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Assay of an Activity in the Serum of Patients with Disorders of Thrombopoiesis That Stimulates Formation of Megakaryocytic Colonies

Cited by 113 publications

References 22 publications

Serum thrombopoietin levels in patients correlate inversely with platelet counts during chemotherapy-induced thrombocytopenia

Serum thrombopoietin levels in patients correlate inversely with platelet counts during chemotherapy-induced thrombocytopenia

Suppression of in vitro megakaryocyte production by antiplatelet autoantibodies from adult patients with chronic ITP

Purification and partial characterization of a megakaryocyte colony-stimulating factor from human plasma.

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