Abstract
The relationship between thrombopoietin (TPO) and its receptor cMpl in thrombocytopenic conditions has not been entirely clarified. To elucidate this interplay may expand the spectrum of indications of TPO mimetics. In this study we have explored the relationship between TPO and cMpl in platelets and megakaryocytes of 43 patients with thrombocytopenia due to idiopathic thrombocytopenic purpura (ITP), bone marrow hypoplasia, myelodysplastic syndromes (MDS), and familial thrombocytopenia. Data were compared to cMpl and TPO in patients with a normal platelet count and in patients with thrombocytosis due to essential thrombocythemia (ET). All but familial patients showed higher TPO compared to controls. All thrombocytopenic states were invariably associated with increased expression of platelet cMPL compared to healthy controls. ET patients showed normal TPO and a trend toward a reduced cMpl expression. Immunofluorescence of bone marrow sections from patients with ITP and MDS failed to show a peculiar pattern compared to controls. Multiple mechanisms regulate TPO and cMpl in thrombocytopenic conditions.
Highlights
Megakaryopoiesis is a multistep process driving the proliferation, differentiation and maturation of hematopoietic stem cells to the generation of platelets, anucleate blood components with a fundamental role in the hemostatic process.[1] platelet count between 20 and 80×109/L
Serum samples were obtained from 15 patients with chronic idiopathic thrombocytopenic purpura (ITP), 8 patients with hypoplastic BM due to aplastic anemia
All samples were treated for double pg/mL) (Figure 1). Both ITP and myelodysplastic syndromes (MDS) patients staining by co-incubation at 4°C overnight with rabbit polyclonal antibody anti cMpl-TpoR (Millipore Upstate) and mouse anti CD41 ly (SZ22 clone – Beckman Coulter Immunotech) monoclonal antibody. o Biotin conjugate secondary antibody followed by streptavidin/Texas Red was used for e rabbit cMpl, whereas secondary antimouse s FITC antibody was used for mouse anti CD41. u In order to assess specificity of the primary antibody, in a separate session primary l immunoreaction was performed by incubating ia (1 h, RT) samples with goat polyclonal antibody c anti cMpl mapping the N-terminus of the pror tein (1:10; Santa Cruz Biotechnology, CA, USA), e followed by incubation (ON, 4°C) with rabbit polyclonal antibody anti cMpl-TpoR
Summary
Serum samples were obtained from 15 patients with chronic ITP, 8 patients with hypoplastic BM due to aplastic anemia 2014 Licensee PAGEPress, Italy Hematology Reports 2014; 6:4996 doi:10.4081/hr.2014.4996
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