title: Immune thrombocytopenic purpura (ITP) plasma and purified ITP monoclonal autoantibodies inhibit megakaryocytopoiesis in vitro.
abstract: To determine if megakaryocytes are targeted by immune thrombocytopenic purpura (ITP) autoantibodies, as are platelets, we have studied the effects of ITP plasma on in vitro megakaryocytopoiesis. Umbilical cord blood mononuclear cells were incubated in the presence of thrombopoietin and 10% plasma from either ITP patients (n = 53) or healthy donors. The yield of megakaryocytic cells, as determined by flow cytometry, was significantly reduced in the presence of ITP plasma containing antiplatelet glycoprotein Ib (GPIb) autoantibodies (P <.001) as compared with both the control and patient plasma with no detectable anti-GPIIb/IIIa or anti-GPIb autoantibodies. Platelet absorption of anti-GPIb autoantibodies in ITP plasmas resulted in double the megakaryocyte production of the same plasmas without absorption, whereas platelet absorption of control plasma had no effect on megakaryocyte yield. Furthermore, 2 human monoclonal autoantibodies isolated from ITP patients, 2E7, specific for human platelet glycoprotein IIb heavy chain, and 5E5, specific for a neoantigen on glycoprotein IIIa expressed on activated platelets, had significant inhibitory effects on in vitro megakaryocytopoiesis (P <.001). Taken together, these data indicate that autoantibodies against either platelet GPIb or platelet GPIIb/IIIa in ITP plasma not only are involved in platelet destruction, but may also contribute to the inhibition of platelet production.
authors.affiliation: Hemostasis and Thrombosis, Children's Hospital of Orange County, 455 S Main St, Orange, CA 92868, USA. mchang@choc.org
authors.forename: Mei
authors.initials: M
authors.lastname: Chang
authors.forename: Peggy A
authors.initials: PA
authors.lastname: Nakagawa
authors.forename: Shirley A
authors.initials: SA
authors.lastname: Williams
authors.forename: Michael R
authors.initials: MR
authors.lastname: Schwartz
authors.forename: Karen L
authors.initials: KL
authors.lastname: Imfeld
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Buzby
authors.forename: Diane J
authors.initials: DJ
authors.lastname: Nugent
mesh_terms: D000293:Adolescent; D000911:Antibodies, Monoclonal / Q000097:blood / Q000494:pharmacology; D001323:Autoantibodies / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002648:Child; D002675:Child, Preschool; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D007223:Infant; D008297:Male; D008533:Megakaryocytes / Q000166:cytology* / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000276:immunology; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*; D016553:Purpura, Thrombocytopenic, Idiopathic / Q000276:immunology* / Q000473:pathology; D034061:Thrombopoiesis / Q000187:drug effects*
chemical_list: D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D010980:Platelet Membrane Glycoproteins