The Examination Method of Glycoprotein CD62P and CD63 on Platelet Membrane and Their Expression in Adults Patients with Diabetes Mellitus / 中国实验血液学杂志

ABSTRACT

To study the optimal examination method of CD62P and CD63 and investigate platelet activation in patients with diabetes mellitus (DM), whole blood labeled directly with monoclonal antibodies CD62P and CD63 and flow cytometry were used to evaluate the positive percentages and the mean fluorescence intensity of CD62P and CD63. The specimens of peripheral blood obtained from 10 healthy adults were divided into two groups. In the unfixing group, the positive percentages of CD62P and CD63 at the periods of 30, 60, 90 and 120 minutes after staining were (7.57 +/- 2.33)%, (20.50 +/- 5.70)%, (28.70 +/- 5.67)% and (36.52 +/- 6.13)%, and, (0.89 +/- 0.36)%, (1.11 +/- 0.84)%, (2.35 +/- 2.02)% and (5.43 +/- 3.66)% respectively, their respective MFI were 1.57 +/- 0.13, 1.88 +/- 0.08, 2.00 +/- 0.09 and 2.38 +/- 0.22 and 3.91 +/- 0.11, 4.07 +/- 0.16, 4.38 +/- 0.14 and 4.44 +/- 0.19. However, in fixing group with 1% paraformaldehyde, the results had not any obvious change and almost were same. Besides it, the positive percentages of CD62P and CD63 in 37 adult patients with DM were (14.11 +/- 6.68)% and (2.71 +/- 1.74)%, significantly higher than that in the normal controls. It is concluded that the CD62P and CD63 on platelet membrane were very sensitive and would be easily activated in vitro, all manipulations that includes labeling with antibody, incubation and detection using flow cytometry should be finished within 30 minutes after samples collected. While fixing by using 1% paraformaldehyde can steady the labeling compounds and effectively prevent the artificial activation of platelet, and keep the stable results within two hours after the samples labeled. In adult patients with DM, the relationship between the cardiovascular complication of diabetes and platelet activation might be existed.