血小板活化功能及血小板微粒对重型血友病A临床异质性的意义

Significance of platelet activating capacity and platelet-derived microparticles in clinical phenotype heterogeneity of hemophlia A: report of 31 cases

目的应用流式细胞术检测血小板表面特异性抗原和血小板微粒(platelet microparticles,PMPs),并探讨血小板活化程度及PMPs对重型血友病A患者临床表现异质性的意义。方法应用特异性荧光单抗CD41a-APC,CD42b-PE,CD61-FITC,CD62P-PE,and CD63-PE及标准荧光校准微珠流式细胞术检测血小板微粒,比较不同临床表型重型血友病A患者(FⅧ∶C<2%)外周血血小板CD62P和CD63表达率和PMPs水平,其中轻度表型10例(年出血次数<6),重度表型21例(年出血次数≥24),正常对照者17例。结果重度表型组和轻度表型组CD62P阳性率均值高于正常对照组[(36.2±9.27)%vs(22.06±10.72)%,P=0.001];[(49±11.81)%vs(22.06±10.72)%,P=0.000];轻度表型组CD62P表达阳性率高于重度表型组[(49±11.81)%vs(36.2±9.27)%,P=0.0408)]。重度出血表型组和轻度出血表型组CD63阳性率均值高于正常对照组[(34.82±7.13)%vs(19.08±17.05)%,P=0.04;(51.15±14.30)%vs(19.08±17.05)%,P=0.000];轻度表型组较重度表型组CD63阳性率均值高[(51.15±14.3)%vs(34.82±7.13)%,P=0.004 4]。轻度表型组PMPs高于重度表型组及正常对照组高[(2.06±1.33)%vs(0.95±0.64)%,P=0.02];[(2.06±1.33)%vs(1.06±1.20)%,P=0.02]。结论血小板活化功能及血小板微粒是影响重型血友病A临床表型的因素,轻度表型患者血小板活化功能、血小板微粒水平较重度表型高。

Objective To determine the levels of platelet specific antigens and platelet-derived microparticle（ PMPs） by flow cytometry（ FCM） in hemophlia A patients, and analyze their clinical significances in the phenotype heterogeneity of the patients with severe hemophlia A. Methods Thirty-one patients with severe hemophlia A（ aged from 18 to 33,median 23. 5） were enrolled in this study. They were 10 with mild type（ annualized bleed rates 〈 6） and 21 with moderate and severe type（ annualized bleed rates ≥24）. Another 17 healthy male volunteers served as normal controls. Their supernatant plasma samples were measured for baseline PMPs count, with phycoerythrin（ PE）-conjugated monoclonal antibodies CD41.Fluorescent microspheres of 1. 0 μm and standard fluorescent microbeads（ 0 to 1. 0 μm） in diameter were used to set the microparticle gate,and the antibodies of CD41a-APC,CD42b-PE,CD61-FITC,CD62P-PE,and CD63-PE specific to platelets derived from platelets were employed to analyze the expression of corresponding platelet activating antigens by flow cytometry. Results The expression percentage of CD62 P and CD63 was obviously higher in the mild type than the sever type patients（ mean ± SD： 49 ± 11. 81 vs 36. 2 ± 9. 27,P =0. 040 8）;（ mean ± SD： 51. 15 ± 14. 3 vs 34. 82 ± 7. 13,P = 0. 004 4） and a higher mean level of PMPs counts in mild type patients compared to severe type（ mean ± SD： 2. 06 ± 1. 33 vs 0. 95 ± 0. 64,P = 0. 02）.Conclusion Platelet activating capacity and PMPs exert certain effect on clinical phenotypes of hemophlia A.The mild patients have stronger platelet activating capacity and higher PMPs count than the severe ones.