Effects of EDTA on the membrane glycoproteins IIB-IIIa complex — analysis using flow cytometry

Nomura, Shosaku; Nagata, Hirokazu; Oda, Kinichiro; Kokawa, Terutoshi; Yasunaga, Kōjirō

doi:10.1016/0049-3848(87)90239-8

Cited by 61 publications

(30 citation statements)

References 26 publications

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“…As an index of platelet activation, CD62P expression was quantified by immunostaining with anti-CD62P monoclonal antibody (CLB-thromb/6, Immunotech, Marseille, France). Platelet expression of GPIIb/ IIIa and of GPIb was analysed with anti-GPIIb/IIIa (NNKY1±32) [33,36] and anti-GPIb (NNKY5±5) [37] monoclonal antibodies, respectively.…”

Section: Methodsmentioning

confidence: 99%

Detection of monocyte-derived microparticles in patients with Type II diabetes mellitus

et al. 2002

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Patients with diabetes mellitus develop hypercoagulability, platelet hyperaggregability and premature atherosclerosis [1±4]. Atherosclerosis is the major cause of death in patients with diabetes mellitus, leading to a high mortality rate in all forms of the disease [5]. Classical risk factors, including hyperlipidaemia, hypertension, and obesity, do not completely account for the increased incidence of atherosclerosis associated with diabetes [6]. Recent reports suggest that increased expression of endothelial adhesion Diabetologia (2002) Results. The concentration of MDMPs in diabetic patients was higher than in normal subjects. We found no differences in the binding of anti-GPIIb/IIIa and anti-GPIb monoclonal antibodies between groups. There were differences, however, in the concentrations of PDMPs, plt-CD62P, and plt-CD63 between Type II (non-insulin-dependent) diabetes mellitus patients and control subjects (PDMPs: 585 25 vs 263 9, p < 0.01; plt-CD62P: 28.1 % 1.4 % vs 9.4 % 0.6 %, p < 0.001; plt-CD63: 28.1 % 1.4 % vs 8.6 % 0.5 %, p < 0.001). Amounts of MDMPs correlated positively with these platelet activation markers, and the relation between PDMP and MDMP was the most significant. The concentration of MDMP in patients who had diabetes complicated with nephropathy, retinopathy, or neuropathy was higher than in those without diabetes-related complications. The increase in MDMP was particularly significant in patients with nephropathy. Concentrations of sE-selectin were higher in Type II diabetes patients than in control subjects, and correlated with MDMP, PDMP, plt-CD62P, and plt-CD63 levels in nephropathy patients. Conclusion/interpretation. In Type II diabetes patients, we detected increased activation of monocytes, which could have been stimulated by activated platelets and PDMPs. Because the activation of monocytes is associated with vascular endothelial damage, high concentrations of MDMPs could indicate vascular complications in diabetes patients, especially those who have diabetes-related nephropathy. [Diabetologia (2002) 45:550±555]

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Section: Methodsmentioning

confidence: 99%

Detection of monocyte-derived microparticles in patients with Type II diabetes mellitus

et al. 2002

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“…As an index of platelet activation, CD62P expression was quantified by immunostaining with an anti-CD62P monoclonal antibody (CLB-thromb/6, Immunotech, Marseille, France), CD63 (Immunotech), PAC-1 (Becton Dickinson, CA, USA) ( Figure 1). Platelet expression of glycoprotein (GP) IIb/IIIa and GPIb was detected with an anti-GPIIb/IIIa monoclonal antibody (NNKY1-32) 33 and an anti-GPIb monoclonal antibody (NNKY5-5).…”

Section: Flow Cytometry Of Activated Platelets and Microparticlesmentioning

confidence: 99%

Effects of efonidipine on platelet and monocyte activation markers in hypertensive patients with and without type 2 diabetes mellitus

2002

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We compared the levels of microparticles, platelet activation markers, soluble cell adhesion molecules, and soluble selectins between hypertensive patients with and without type 2 diabetes and control subjects. Binding of anti-glycoprotein IIb/IIIa and anti-glycoprotein Ib monoclonal antibodies to platelets did not differ significantly between the hypertensive patients and controls, but platelet expression of activation markers (CD62P, CD63, PAC-1, and annexin V) was higher in the hypertensive patients. Platelet-derived microparticle (PDMP) and monocyte-derived microparticle (MDMP) levels were significantly higher in the hypertensive patients than in the controls. Soluble ICAM-1, VCAM-1, P-selectin, and E-selectin levels were also higher in the

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“…Monoclonal Antibodies The following monoclonal antibodies were used: antiGPIIb-IIIa (NNKY1-32) (21,22), anti-GPIIIa (SZ21; Cosmo Bio, Tokyo) (29), anti-GPIb (NNKY5-5) (22), and anti-fibrinogen (Chemicon International, Inc., CA). These antibodies were pure as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).…”

Section: Methodsmentioning

confidence: 99%

“…Samples were assayed on a Becton Dickinson FACS Research Analyzer using the 50 pm orifice (21,22). The instrument was calibrated for fluorescence and light scatter using 2.0 pm beads (Becton Dickinson and Co., Oxnard CA).…”

Section: Flow Cytometric Analysismentioning

confidence: 99%

“…It has recently been reported that GPIIb and GPIIIa exist predominantly as a Ca2+ -dependent, heterodimer complex, and that incubation of intact platelets with ethylenediamine tetraacetic acid (EDTA) at 37°C causes the progressive dissociation of such GPIIb/IIIa complexes (6,21,26,27). The binding of GPIIb/IIIa to fibrinogen is detected only after the stimulation of platelets with agonists such a s ADP or thrombin (14,181.…”

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Differences between platelet and microparticle glycoprotein IIb/IIIa

et al. 1992

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Glycoprotein (GP) IIb and IIIa are major constituents of the platelet membrane which are involved in forming the fibrinogen receptor on activated platelets. We used flow cytometry to study the effects of ethylene‐diamine tetraacetic acid (EDTA) on the membrane GPIIb/IIIa complexes of platelets and microparticles, and to study the effects of cations on dissociated GP complexes. Microparticles were detected by both the volume signal and by fluorescence using an FITC‐conjugated anti‐GPIb antibody (NNKY5–5). When platelets were stimulated with ADP, calcium ionophore A23187, or thrombin, fibrinogen binding to the platelet surface increased markedly. However, fibrinogen binding to microparticles showed little increase in response to such agonists. Microparticle GPIIb/IIIa complexes were dissociated by incubation with EFTA at 37°C but did not reassociate after treatment with divalent cations (Ca2+, Mg2+, and Mn2+) in contrast to platelet GPIIb/IIIa complexes. These results suggest that some interaction of GPIIb/IIIa and linked structures like the platelet cytoskeleton may be involved in the reassociation of dissociated GPIIb and GPIIIa, perhaps explaining the failure of reassociation of microparticle GPIIb/IIIa (i.e., the fibrinogen binding to microparticles). © 1992 Wiley‐Liss, Inc.

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Effects of EDTA on the membrane glycoproteins IIB-IIIa complex — analysis using flow cytometry

Cited by 61 publications

References 26 publications

Detection of monocyte-derived microparticles in patients with Type II diabetes mellitus

Detection of monocyte-derived microparticles in patients with Type II diabetes mellitus

Effects of efonidipine on platelet and monocyte activation markers in hypertensive patients with and without type 2 diabetes mellitus

Differences between platelet and microparticle glycoprotein IIb/IIIa

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