In part supported by the Bundesministerium für Jugend, Frauen, Familie, and Gesundheit, Bonn, Federal Republic of Germany, the Minister für Wissenschaft und Forschung, Nord-Rhein-Westfalen, Duesseldorf, Federal Republic of Germany, and the Paul-Kuth-Stiffung, Wuppertal, Federal Republic of Germany.
Article first published online: 30 MAR 2005
Copyright © 1990 Wiley-Liss, Inc.
Volume 11, Issue 5, pages 652–656, 1990
How to Cite
Tschoepe, D., Spangenberg, P., Esser, J., Schwippert, B., Kehrel, B., Roesen, P. and Gries, F. A. (1990), Flow-cytometric detection of surface membrane alterations and concomitant changes in the cytoskeletal actin status of activated platelets. Cytometry, 11: 652–656. doi: 10.1002/cyto.990110515
In part presented at the 3d Erfurt Workshop on Platelets, Erfurt, German Democratic Republic, 26–30.6.1989.
- Issue published online: 21 JUN 2005
- Article first published online: 30 MAR 2005
- Manuscript Accepted: 24 FEB 1990
- Manuscript Received: 11 JUL 1989
- Activation markers;
- actin status
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.