内皮细胞及组织因子在GVHD中作用机制的初步研究

本研究检测人内皮细胞组织因子(TF)和IL-2的表达水平,以探讨使用免疫抑制剂后内皮细胞TF及IL-2表达水平的变化及内皮细胞损伤和TF异常表达在GVHD中的意义。体外混合人内皮细胞与异体人淋巴细胞及加入不同浓度的CsA共培养4-6小时后,用流式细胞术和RT-PCR检测内皮细胞中TF的表达水平,应用ELISA法检测上清中IL-2的水平。结果表明:体外混合人内皮细胞与异体人淋巴细胞共培养后,与未混合培养的相比内皮细胞表达TF增加(p0.01),并且在培养液中IL-2水平也相应增高(p0.01),在加入不同浓度的CsA之后,TFmRNA表达下降,相应的培养液中IL-2浓度也下降(p0.01)。结论:内皮细胞损伤与TF异常表达在骨髓移植后的GVHD中起关键的作用,其中受体的内皮细胞受到供体淋巴细胞刺激后高表达TF可能是GVHD发生、发展过程中的关键步骤。

Preliminary Study on Role of Endothelial Cells and Tissue Factor in Graft-versus-Host Disease

This study was purposed to detect the expression level of human endothelial cell tissue factor（hECTF） and concentration of IL-2,and to investigate the alterations of hECTF and IL-2 after using immunosuppressive agent（cyclosporin,CsA） and explore the significance of endothelial cell（EC） lesion and abnormal expression of tissue factor（TF） in GVHD.Human endothelial cells and allogeneic lymphocytes were mixed and cultured as well as were coculatured with CsA for 4-6 hours in vitro,then the expression level of hECTF was detected by flow cytometry and RT-PCR,the concentration of IL-2 in supernatant was assayed by ELISA.The experiment was divided into 4 groups： 1st group — non-mixed-cultured group（negative control group）,2nd group — mixed-cultured group（positive control group）,3rd group — mixed-cocultured group with 1 μg/ml CsA and 4th group — mixed-cocultured group with 2 μg/ml CsA.The results showed that as compared with non-mixed-cultured group（negative control group）,the expression level of hECTF and concentration of IL-2 in another 3 groups significantly increased（p0.01）,while as compared with positive control group,the expression level of hECTF and concentration of IL-2 in cocultured groups with CsA both decreased（p0.01）.It is concluded that the lesion of EC and abnormal expression of TF play a crucial role in GVHD,among which the high expression of TF after being stimulated by donor＇s lymphocytes may be the key step for occurence and progression of GVHD.