IMMUNOLOGICAL METHODS FOR EVALUATION OF PLATELET FUNCTIONS IN THE PATIENTS WITH CARDIOVASCULAR DISEASES

Abstract

The aim of this work was a simultaneous analysis of functional platelet activity using both standard optical aggregometry and flow cytometry, by means of measuring P-selectin and GP IIb-IIIa glycoprotein receptor contents on the surface of ADP-induced platelets. GP IIb-IIIa amounts and P-selectin expression on cell surface are significantly increased on platelets after activation with 10 mcM ADP, both in normal donors and in aspirin-treated patients. In clopidogrel-treated patients, the amounts of expressed P-selectin and GP IIb-IIIa did not change after ADP induction, thus suggesting absence of detectable ADP-induced platelet activation in this group. By means of Spearman criterion, a distinct correlation was found between the P-selectin expression, GP IIb-IIIa contents, and rates of standard ADP-induced aggregation. A combination of standard ADP-induced aggregation/activation of platelets with flow cytometry using FITC or PE-labeled antibodies to P-selectin and GP IIb-IIIa, allowed us to apply immunological approaches to analysis of platelet hemostasis, to propose novel laboratory criteria for evaluation of platelet hyperaggregation, and to detect differential effects of clopidogrel and aspirin.

Keywords

• p-selectin expression
• gp iib-iiia
• adp-induced platelet aggregation
• flow cytometry