Effect of a New Monoclonal Anti-Glycoprotein IX Antibody, KMP-9, on High Shear-Induced Platelet Aggregation

Miyake, Tetsuya; Nomura, Shosaku; Komiyama, Yutaka; Miyazaki, Yasuhiko; Kagawa, Hideo; Masuda, Midori; Takahashi, Hakuo; Fujimura, Yoshihiro; Ikeda, Yasuo; Fukuhara, Shirou

doi:10.1055/s-0038-1657650

Cited by 33 publications

(14 citation statements)

References 27 publications

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“…Both intact and aggregated platelets were removed by centrifugation at 1000 g for 15 min to yield a supernatant containing microparticles only. Ten microlitres of washed intact platelets (3 10 8 /ml) was then added to the supernatant and the mixture was incubated with KMP-9 [35] (a FITC-labelled monoclonal antibody against platelet GPIX) for 30 min in the dark at room temperature. After incubation, samples were diluted 1:10 with HEPES-Tyrode's buffer containing 5 nmol/l EGTA and analysed with an Ortho Cytoron Absolute Analyzer (Ortho Diagnostic Systems, Tokyo, Japan).…”

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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“…The washed intact platelets (10 l, 3 × 10 8 /ml) were then added to the supernatant, and incubation with KMP-9 32 (a fluorescein isothiocyanate (FITC)-labelled monoclonal antibody directed against platelet GPIX) was performed for 30 min in the dark at room temperature. After incubation, samples were diluted 1:10 with Hepes-Tyrode's buffer containing 5 nmol/l EGTA and analysed with an Ortho Cytoron Absolute Analyzer (Ortho Diagnostic Systems, Tokyo, Japan).…”

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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“…4-(2-Hydroxyethyl)-1-piperazine-ethanesulfonic acid (Hepes) and prostaglandin E 1 (PGE 1 ) were purchased from Wako Chemicals (Tokyo, Japan). The following monoclonal antibodies were used for this study: CD41 (anti-· IIb ß 3 complex; NNKY2-11) [28], CD42a (KMP-9) [29], CD42b (NNKY5-5) [18] and CD9 (NNKY1-19) [30] were our antibodies. CD31, CD32, CD62P and CD63 were obtained from Immunotech (Marseille, France).…”

Section: Methodsmentioning

confidence: 99%

Morphological Differences between GPIb Antibody-Induced and Shear Stress-Induced Platelet Aggregates

Nomura

Tandon

Nakamura

et al. 2000

Pathophysiol Haemos Thromb

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We determined the morphological differences between NNKY5-5-induced and shear stress-induced platelet aggregates using confocal laser scanning microscopy, and investigated the effects of cytokines on these aggregates. As shear stress was increased from low to high, many aggregates were generated. Some platelets and aggregates already exhibited PAC-1 binding at low shear stress. And finally, aggregates were clearly stained by PAC-1 at high shear stress. Low shear stress-induced aggregates included fibrinogen, but not von Willebrand factor (vWF). In contrast, high shear stress-induced aggregates included both fibrinogen and vWF. NNKY5-5-induced platelet aggregates exhibited both PAC-1 binding and fibrinogen, but vWF was not found in them. The aggregate formation in NNKY5-5-treated platelet-rich plasma (PRP) at low shear stress was clearly more pronounced than that in untreated PRP. In addition, aggregates in NNKY5-5-treated PRP formed after 6 min under low shear stress were clearly stained by PAC-1 and included fibrinogen, but vWF was not found in them. In order to investigate the effects of cytokines on platelet activation under NNKY5-5 stimulation, changes in light transmission and light scatter were assessed with an AG-10. G-CSF, IL-6 and thrombopoietin (TPO) each enhanced light scatter compared to control PRP, although IL-3, GM-CSF, erythropoietin and stem cell factor did not. In particular, TPO significantly enhanced the ‘%-T’ and ‘S-Max’ of NNKY5-5-treated PRP. TPO clearly enhanced the aggregate formation with high shear stress or NNKY5-5 stimulation. These results suggest that the glycoprotein Ib (GPIb)-unmediated aggregates can be formed with only fibrinogen, but the GPIb-mediated aggregates by vWF and fibrinogen synergistically. In particular, the latter is formed more firmly, and both aggregates are enhanced by TPO.

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Effect of a New Monoclonal Anti-Glycoprotein IX Antibody, KMP-9, on High Shear-Induced Platelet Aggregation

Cited by 33 publications

References 27 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

Morphological Differences between GPIb Antibody-Induced and Shear Stress-Induced Platelet Aggregates

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