【摘要】 目的　观察再生障碍性贫血 (AA)患者骨髓中肿瘤坏死因子 (TNF α)含量变化、分泌的细胞及临床意义。方法　应用Elisa检测骨髓TNF α水平 ,多色免疫荧光标记技术和流式细胞术分析T细胞膜型、胞浆内TNF α含量 ;纯化T细胞经植物凝血素 (PHA)、Flt3配体 (FL)及白介素 2 (IL 2 )激发后培养上清中TNF α水平 ;T辅助细胞 (Th)亚群改变。结果　AA患者骨髓中TNF α水平显著升高与对照组比P <0 0 5 ,且与患者外周血白细胞数呈负相关 (r=0 78,P <0 0 5 )。患者骨髓T细胞无膜型TNF α及受体表达 ,CD4 + 和CD8+ 细胞胞浆中TNF α含量显著增加 ,但两者无差别 ;淋巴细胞经PHA、FL及IL 2激发后培养上清中TNF α水平升高 ,尤以PHA的作用最为显著 ;AA患者骨髓中T细胞极化异常 ,Th1细胞比例增多 ,Th1/Th2增高。结论　AA患者淋巴细胞极化异常 ,Th1细胞比例增多 ,骨髓中T细胞胞浆中TNF α含量显著增多是AA骨髓衰竭的原因之一。更多还原

【Abstract】 Objective To investigate the changes of tumor necrosis factor-α(TNF-α),cell source and its clinical significance in patients with aplastic anemia(AA).Methods The concentrations of TNF-α in bone marrow of AA patients were determined by Elisa assay.Single or double staining immunofluorescence assay and flow cytometry were used to trace the TNF-α producing cell and the expression of its receptor.The levels of TNF-α were measured when the T lymphocytes were cocultured with phytohemagglutinin(PHA),flt3 ligand(FL)and interleukin-2(IL-2)for 3 days.The changes of T help subsets in those patients were also investigated.Results The levels of TNF-α in bone marrow of AA patients increased significantly in comparison with controls(P<0.05),which correlated inversely to the number of white blood cells in peripheral blood(r=-0.78,P<0.05).Flow cytometry analysis showed that there was no TNF-α or its receptor expresion on T cells.The intracellular expression of TNF-α increased significantly both in CD4 + and CD8 + T cells.But there was no difference between them.The levels of TNF-α supermatant increased significantly when T cells cocultured with PHA,FL and IL-2 for 3 days,especially in those incubated with PHA.Our results indicated there was an abnormal polarization of T cells in AA patients.The ratio of Th1 to Th2 increased in patients with AA.Conclusion Abnormal T cell polarization,increasing Th1 cells and overexpression of intracellular TNF-α in T cell of AA patients were one of main reasons causing bone marrow failure.更多还原