Darstellung von Thrombozytenmembranproteinen mit monoklonalen Antikörpern im durchflußzytometrischen Bio-Assay

Tschöpe, D; Schauseil, S; Roesen, P.; Kaufmann, L.; Gries, F. A.

doi:10.1007/bf01774225

Cited by 14 publications

(2 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In order to overcome this handicap, we developed a flow cytometric bioasy to characterize the functional status of freshly fixed platelets directly platelet by platelet. 74 ' 75 After activation, platelets un dergo a complex downstream molecular cell-processing cascade. In the very early steps, concomitant with the polymerization of cytoskeletal proteins, the antigenicity of the outer membrane changes due to the exposure of subcellular epitopes, which are not detectable in the rest ing state.…”

Section: Platelet Activationmentioning

confidence: 99%

Platelets in Diabetes: The Role in the Hemostatic Regulation in Atherosclerosis

Tschöepe

Roesen²,

Schwippert³

et al. 1993

Semin Thromb Hemost

View full text Add to dashboard Cite

Vascular diseases and related complications still represent the main cause of death in diabetic patients. Neuropathy, nephropathy, retinopathy, and disturbed nutritive tissue perfusion may result from reduced capillary microcirculation. These disturbances are diabetes specific. Macroangiopathy does not differ structurally from atherosclerotic lesions of nondiabetic subjects, but leads to accelerated cerebral, coronary, and peripheral artery disease. Occurrence of life-terminating thrombotic events, which are superimposed on those vascular lesions, are increased. Thus, morbidity and mortality of diabetics depend mainly on vascular complications. Normal blood flow is a prerequisite of adequate organ perfusion and results from vasomotion, plasma components, corpuscular blood elements, vascular architecture, and the undisturbed interaction of these components at the endothelial interface. Functional thromboresistance of the endothelial layer is reduced in the diabetic state. Increased intravascular thrombin generation, reduced fibrinolytic potential, and hyperactive platelets lead to a prethrombotic state. This thrombotic diathesis increases the permanent danger of acute flow interruption. Activated platelets operate by three mechanisms: (1) Microembolization of the capillaries; (2) local progression of preexisting vascular lesions by secretion of constrictive, mitogenic, and oxidative substances; (3) trigger of the prognosis-limiting arterial thrombotic event. We were able to show that the increased functional properties of diabetic platelets result from the primary release of larger platelets with enhanced thromboxane formation capacity and increased numbers of functional glycoprotein receptors GPIb and GPIIb/IIIa, which are synthesized in the megakaryocytes. The megakaryocyte-platelet system is turned on in diabetes mellitus. It could be demonstrated with the Duesseldorf III method of flow cytometric activation marker testing (CD62, CD63, thrombospondin) that predominantly large platelets circulate in an activated state in diabetes mellitus.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Section: Platelet Activationmentioning

confidence: 99%

Platelets in Diabetes: The Role in the Hemostatic Regulation in Atherosclerosis

Tschöepe

Roesen²,

Schwippert³

et al. 1993

Semin Thromb Hemost

View full text Add to dashboard Cite

show abstract

“…Monozyten und neutrophile Granulozyten exprimieren neben dem P-Selectin Rezeptor PSGL-1 auch verschiedene kohlenhydratartige Strukturen wie das "sialyl Lewis X" Oligosaccharid, über die es zur Bindung an Thrombozyten und Endothelzellen kommt [6,7]. Neben dem an -Granula gebundenen P-Selectin, welches als Nachweis der Aktivierung von Thrombozyten mittels monoklonaler Antikörper im Durchflußzytometer dient [8], existiert auch eine lösliche Form des Proteins (sP-Selectin) in der Zirkulation [9,10], die sich enzymimmunometrisch quantitativ bestimmen läßt [11]. Derzeit ist jedoch noch nicht hinreichend geklärt, ob es sich bei sP-Selectin um intaktes -Selectin mit lediglich fehlender transmembraner Domäne oder um Molekülfragmente handelt [12,13].…”

unclassified

Lösliches P-Selectin und β-Thromboglobulin bei koronarer Herzkrankheit und akutem Myokardinfarkt.

Stiegler¹,

Fischer²,

Schoebel³

et al. 1999

LaboratoriumsMedizin

View full text Add to dashboard Cite

Zusammenfassung: P-Selectin ist ein Adhäsionsmolekül, welches in der Membran der -Granula von Thrombozyten sowie in "Weibel-Palade-Körpern" von Endothelzellen lokalisiert ist. Eine lösliche Isoform von P-Selectin, "soluble P-Selectin u (sP-Selectin), kann enzymirhmunornetrisch in humanem Plasma nachgewiesen werden. P-Selectin wird im Rahmen der "Release-Reaktion" von aktivierten Thrombozyten vermehrt exprimiert, während ß-Thromboglobulin aus thrombozytären -Granula sezerniert wird. Aktivierte Thrombozyten und deren Interaktion mit der Gefäßwand spielen eine entscheidende Rolle in der Pathogenese der korpnaren Herzkrankheit und den akuten Koronarsyndromen wie der instabilen Angina pectoris und dem akuten Myokardinfarkt. Um die Relevanz von sP-Selectin und ß-Thromboglobulin bei koronarer Herzkrankheit zu evaluieren, wurden bei Patienten mit stabiler Angina pectoris (sAP, n=20), instabiler Angina pectoris (iAP, n=12) und akutem Myokardinfarkt (AMI, n=12) sowie bei Normalprobanden (NP, n=ll) die Konzentrationen an sP-Selectin und ß-Thromboglobulin enzymimmunometrisch in der systemischen Zirkulation analysiert. Die 'drei untersuchten Patientengruppen wiesen unabhängig vom klinischen und angiokardiographischen Schweregrad der koronaren Herzkrankheit signifikant erhöhte sP-Selectin Plasmakonzentrationen (Mittelwert±SD) verglichen mit Normalprobanden auf [NP: 98±20 ng/ml vs. s AP: 133±30 ng/ml, p<0,01; vs. iAP: 128±28 ng/ml, p<0,01; vs. AMI: 144±72.ng/ml,p<0,05]. ß-Thromboglobulin Konzentrationen waren lediglich bei Patienten mit akutem Myokardinfarkt signifikant erhöht [NP: 30±20 lU/ml vs. AMI: 39±14 lU/ml, p<0,05]. Bei den Patienten mit akutem Myokardinfarkt fand sich darüber hinaus eine positive Korrelation von sP-Selectin mit der maximalen Aktivität der Creatin-Kinase im Serum [r=0,72, p<0,01].

show abstract

Flow‐cytometric detection of surface membrane alterations and concomitant changes in the cytoskeletal actin status of activated platelets

et al. 1990

Self Cite

View full text Add to dashboard Cite

Occlusive vascular diseases are promoted by a “prethrombotic state” with increased platelet activity. Polymerization of cytoskeletal proteins and exposure of subcellular structures or rebinding of secreted proteins have been characterized as early reactions after platelet activation preceding adhesion and aggregation. Here, we demonstrate the kinetic increase in specific binding of monoclonal antibodies to thrombospondin (P10) and to platelet membrane activation markers CD63 (GP53, a 53 kD lysosomal protein) and CD62 (GMP140, a 140 kD alpha granule protein) by using a flow‐cytometric bio‐assay and the related change in the actin status by using the DNase‐I inhibition assay after stimulation of normal human platelets with 0.2 U/ml thrombin. F‐actin was raised from 41% to 51% of total platelet actin content 30 s after stimulation and remained thereafter constant (50% at 60 s). Simultaneously, the percentage of P10, CD63, and CD62 positive platelets was elevated from 5.4%, 24.4%, and 9.1% to 67.4%, 80.2%, and 82.3% respectively. The mean number of P10, CD63, and CD62 antibody binding sites increased from 3,300, 1,715, and 2,146 to 6,400, 6,800, and 9,016 per platelet. Conclusively, changes in the organization of the cytoskeletal protein “actin” and exposure of subcellular structures indicating platelet secretion can be regarded as markers of early platelet activation. Thus, the parallel response in both analytical systems provides further support for the diagnostic concept of flow‐cytometric detection of preactivated platelets in the peripheral blood by using fluochrome staining procedures detecting activation dependent structural alterations directly at the cellular level.

show abstract

Darstellung von Thrombozytenmembranproteinen mit monoklonalen Antikörpern im durchflußzytometrischen Bio-Assay

Cited by 14 publications

References 17 publications

Platelets in Diabetes: The Role in the Hemostatic Regulation in Atherosclerosis

Platelets in Diabetes: The Role in the Hemostatic Regulation in Atherosclerosis

Lösliches P-Selectin und β-Thromboglobulin bei koronarer Herzkrankheit und akutem Myokardinfarkt.

Flow‐cytometric detection of surface membrane alterations and concomitant changes in the cytoskeletal actin status of activated platelets

Contact Info

Product

Resources

About