急性ITP患者T细胞亚群及血小板凋亡相关蛋白的检测

目的 检测凋亡相关蛋白Fas、FasL在急性特发性血小板减少性紫癜（ITP）患儿T细胞亚群及血小板中的表达，分析其在ITP发病机制中的作用。方法 收集ITP患儿及健康儿童外周抗凝静脉血，分离纯化得T细胞和血小板。以PE标记的CD30 mAb和Cy5标记的CD4、CD8 mAb及FITC标记的Fas、FasL mAb作三色流式细胞术，分析ITP患儿Th／Tc、Thl／Th2、Tc1／Tc2细胞Fas、FasL表达的变化；以FITC标记的Fas、FasL mAb作单色流武细胞术，分析ITP患儿血小板Fas、FasL表达的变化。结果 与健康儿童相比，ITP患儿Th、Th1、Th2及Tc、Tc1、Tc2细胞Fas、FasL表达均显著性升高（P〈O．05）；同时，111P患儿血小板Fas的表达显著性升高（P〈0．05）而FasL的表达无明显变化（P〉0.05）。结论 ITP患儿存在免疫功能失调，T细胞亚群及血小板Fas、FasL表达异常可能与ITP免疫病理机制有关。

Detection of Apoptosis-related Proteins on T Cell Subsets and Thrombocyte in Actue Idiopathic Thrombocytopenic Purpura

Objective To investigate the expression of Fas,FasL on T cell subsets and thrombocyte in patients with idiopathic thrombocytopenic purpura(ITP) and disuss the role of Fas,FasL in ITP pathogenesis.Methods Anti-coagulated venous blood by heparin is collected from idiopathic thrombocytopenic purpura children and healthy children.T cells and thrombocytes are collected by Human CD3+ T cell Enrichment Column.The expression of Fas,FasL on T cell subsets is detected by immunofluorescence staining and tricolor flow cytometry by Fas/CD4,FasL/CD4,Fas/CD8 and FasL/CD8 gating.The expression of Fas,FasL on thrombocytes is made by immunofluorescence straining and mono-color flow cytometry by Fas and FasL gating.Results Compared with healthy children,in ITP children,the expression of Fas,FasL on Th,Th1,Th2,Tc,Tc1 and Tc2 increased significantly(P<0.05).The expression of FasL on Th,Th1 and Th2 increased sharply vs Fas(P<0.05),while the expression of Fas on Tc,Tc1 and Tc2 increased obviously vs FasL(P<0.05).The expression of Fas on thrombocytes increased significantly but the expression of FasL varied little.Conclusion There is immunofunction disorder in ITP children,and the abnormal expression of Fas,FasL on T cell subsets and thrombocytes may involve in ITP immunopathogenesis.