Arteriosclerosis and Thrombosis, Vol 12, 1475-1487, Copyright © 1992 by American Heart Association

ARTICLES

Activation of platelets in blood perfusing angioplasty-damaged coronary arteries. Flow cytometric detection

RE Scharf, A Tomer, UM Marzec, PS Teirstein, ZM Ruggeri and LA Harker
Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, Calif.

Fluorescence-activated flow cytometry has been used to investigate platelet activation in blood flowing through atherosclerotic coronary arteries after sustaining mechanical damage induced by percutaneous transluminal angioplasty (PTCA). For flow cytometry, platelets and platelet-derived microparticles were identified by biotinylated anti- glycoprotein (GP) Ib monoclonal antibody (mAb) and a fluorophore, phycoerythrin-streptavidin. Activated platelets were detected by using a panel of fluoresceinated mAbs specific for activation-dependent platelet epitopes, including 1) activated GPIIb-IIIa complex (PAC1); 2) fibrinogen bound to platelet GPIIb-IIIa (9F9); 3) ligand-induced binding sites on GPIIIa (anti-LIBS1); and 4) P-selectin, an alpha- granule membrane protein expressed on the platelet surface after secretion (S12). The binding of antibodies to platelets was determined in blood that was sampled continuously via heparin-coated catheters from the coronary sinus in 1) patients before, during, and for 30 minutes after PTCA and 2) control patients undergoing coronary angiography without PTCA. Platelets in coronary sinus blood showed significant binding of mAbs that specifically detect activation epitopes associated with the GPIIb-IIIa complex (PAC1, anti-LIBS1, and 9F9) during and for 30 minutes after angioplasty in four of the five patients. The relative proportion of platelets positive for PAC1 and anti-LIBS1 increased from baseline values of 2.0 +/- 0.3% (mean +/- SD) and 2.0 +/- 0.5% to 18 +/- 14% and 28 +/- 14%, respectively, during PTCA or 30 minutes after PTCA (p < 0.01 in both cases). Binding with 9F9 was less prominent. The expression of P-selectin was detected in one of the five patients. By contrast, activation-specific mAbs failed to bind detectably with platelets obtained from 1) the peripheral blood during coronary angiography in eight patients or 2) coronary sinus blood obtained via catheter throughout control catheterization procedures in three patients or before PTCA in five. We conclude that circulating platelets become activated while flowing through PTCA- damaged stenotic coronary arteries and that this process of platelet activation is readily demonstrated by measuring the expression of activation-specific membrane GP epitopes by flow cytometric analysis.

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