Assessing aspirin-induced attenuation of platelet reactivity by flow cytometry.
BACKGROUND
Certain patient subpopulations do not respond to antithrombotic effects of aspirin and different approaches have been proposed to detect and define this so-called aspirin resistance. In this study, a methodological and clinical evaluation of a flow cytometric method for the detection of aspirin-induced inhibition of platelet cyclooxygenase (COX) is presented.


METHODS
Platelet CD62p-antigen (P-selectin) expression was determined by flow cytometry after incubating diluted platelet rich plasma (PRP) with arachidonic acid (ARA). After establishing the method's technical characteristics, it was used to investigate 114 individuals (70 patients with atherosclerotic vascular disease under long-term medication of 100 mg aspirin daily, 29 age-matched patients with vascular disease without anti-platelet medication and 15 healthy volunteers). Data were compared to those obtained by the PFA-100 platelet function analyzer.


RESULTS
Imprecision was between 3.3% and 13%. Sample storage at room temperature increased baseline activity of platelets already after 2 h. After ARA stimulation, the proportion of CD62p-positive platelets was considerably lower in aspirin-treated patients than in controls (median [lower-upper quartile]: 4% [3-6] vs. 50% [29-68], p<0.001). Only one aspirin-treated patient (1.5%) showed normal reactivity to ARA. In contrast to flow cytometry, PFA-100 analysis yielded normal results in 17% of aspirin-treated patients.


CONCLUSIONS
The presented flow cytometric method can be used to monitor aspirin-induced inhibition of platelet COX. Aspirin resistance defined as failure to inhibit platelet COX is a rare phenomenon suggesting that most cases of aspirin resistance detected using the PFA-100 are caused by COX-independent mechanisms.
Hübl
Wolfgang
W
Department of Laboratory Medicine, Wilhelminenspital Vienna, Austria. wolfgang.hiebl@wienkav.at
Assadian
Afshin
A
Lax
Jaqueline
J
Meixner
Ursula
U
Fang
I-Fei
IF
Hagmüller
Georg
G
Panzer
Simon
S
Bayer
Peter M
PM
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D016718:Arachidonic Acid / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology*; D050197:Atherosclerosis; D001792:Blood Platelets / Q000201:enzymology; D016022:Case-Control Studies; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004351:Drug Resistance; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008875:Middle Aged; D019007:P-Selectin / Q000032:analysis; D015539:Platelet Activation / Q000187:drug effects*; D010979:Platelet Function Tests; D011451:Prostaglandin-Endoperoxide Synthases / Q000032:analysis
D016861:Cyclooxygenase Inhibitors; D019007:P-Selectin; D016718:Arachidonic Acid; D011451:Prostaglandin-Endoperoxide Synthases; D001241:Aspirin