Abstract:

Objective To assess the circulating platelet activation and platelet leukocyte adhesion by flow cytometry and lay the foundation for the clinical development of platelet associated diseases. Methods The sodium citrate anti-coagulation blood was collected from 50 cases of healthy volunteers, using CD61/PAC-1/CD62P to detect platelet activation; analysis the percentage of early activation (PAC-1⁺CD62P^-) and late activation (PAC-1⁺CD62P⁺). Moreover, the different time points, such as 0 min, 5 min, 10 min, 15 min, 20 min, 30 min, and 60 min were detected the platelet activation. Via CD61/CD3/CD19/CD11b/CD14/CD45 detected platelet activation, and analyzed the percentages of platelet leukocyte adhesion (CD45⁺CD61⁺), platelet-T lymphocyte adhesion (CD3⁺CD61⁺), platelet-B lymphocyte adhesion (CD19⁺CD61⁺), platelet monocyte adhesion (CD14⁺CD61⁺), platelet-neutrophil adhesion(CD45^low+CD11b⁺ CD61⁺). Results The time point of 15 minutes was detected to establish the reference range for healthy human platelet activation : the percentage of early activation was 3.75 (1.00-9.80) %, late activation was 0.10 (0.00-0.10) %. Detected the percentages of platelet leukocyte adhesion, platelet-T lymphocyte adhesion, platelet-B lymphocyte adhesion, platelet-monocyte adhesion and platelet-neutrophil adhesion showed platelet-monocyte adhesion was the highest proportion (10.75 ± 3.23)%. ConclusionStorage time of blood samples should not exceed 15 min to detect platelet activation. Platelet-monocyte adhesion was the key role in platelet activation analysis. Establishment the fast, stable, accurate evaluation method of platelet activation is important.

Key words: circulating platelet activation, platelet leukocyte adhesion, flow cytometry