title: Endothelial microparticles induce formation of platelet aggregates via a von Willebrand factor/ristocetin dependent pathway, rendering them resistant to dissociation.
abstract: Endothelial microparticles (EMP) released from activated or apoptotic endothelial cells (EC) are emerging as useful markers for detection of EC dysfunction. Our recent observation that EMP carry von Willebrand factor (vWf) led us to investigate their interaction with platelets. EMP were incubated with normal washed platelets in the presence or absence of ristocetin, then platelet aggregates were measured by flow cytometry. In the absence of ristocetin, negligible EMP conjugated with platelets (< 5%) but in the presence of ristocetin (1 mg mL(-1)), EMP induced up to 95% of platelets to aggregate. EMP-platelet interaction was 80% blocked by anti-CD42b, or by 0.1 microm filtration to remove EMP. Platelet aggregates induced by normal plasma or high molecular weight vWf (Humate-P) dissociated 50% within 15-25 min following 1:20 dilution. In contrast, aggregates formed with EMP persisted two- to threefold longer with the same treatment, indicating greater stability. A similar degree of prolongation of dissociation was observed using plasma from thrombotic thrombocytopenic purpura (TTP) patients compared with normal plasma. Addition of EMP to plasma from severe von Willebrand disease restored his ristocetin-induced platelet aggregation. Multimer analysis of vWf on EMP showed unusually large vWf (ULvWf). In summary, EMP carries ULvWf multimers, promote platelet aggregates, and increase the stability of the aggregates thus formed.
authors.affiliation: Wallace H Coulter Platelet Lab., Department of Medicine, University of Miami School of Medicine, Miami, FL 33136, USA. wjy@med.miami.edu
authors.forename: W
authors.initials: W
authors.lastname: Jy
authors.forename: J J
authors.initials: JJ
authors.lastname: Jimenez
authors.forename: L M
authors.initials: LM
authors.lastname: Mauro
authors.forename: L L
authors.initials: LL
authors.lastname: Horstman
authors.forename: P
authors.initials: P
authors.lastname: Cheng
authors.forename: E R
authors.initials: ER
authors.lastname: Ahn
authors.forename: C J
authors.initials: CJ
authors.lastname: Bidot
authors.forename: Y S
authors.initials: YS
authors.lastname: Ahn
mesh_terms: D002478:Cells, Cultured; D019281:Dimerization; D004730:Endothelium, Vascular / Q000737:chemistry*; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry*; D010974:Platelet Aggregation; D011485:Protein Binding; D011697:Purpura, Thrombotic Thrombocytopenic / Q000097:blood; D012310:Ristocetin / Q000378:metabolism* / Q000494:pharmacology; D014842:von Willebrand Diseases / Q000097:blood; D014841:von Willebrand Factor / Q000032:analysis / Q000378:metabolism*
chemical_list: D046911:Macromolecular Substances; D014841:von Willebrand Factor; D012310:Ristocetin