- 學位論文
抗磷脂質抗體對內皮細胞的影響—栓塞可能的機轉
The impact of antiphospholipid antibodies on endothelial cells —A possible mechanism for thrombosis
摘要
目標：探討抗磷脂質症候群的主要症狀－栓塞可能的機轉，抗磷脂質抗體對內皮細胞的影響。 方法：從3個正常人血清、12個Anti-phospholipid Syndrome(APS) 病人血清、3株aCL hybridoma單株抗體純化的IgG和Human umbilical cord vein endothelial cells (HUVECs)共同培養後，以流式細胞儀偵測HUVEC表面所表現的Thrombomodulin(TM)、Tissue Factor (TF)、ATP Diphosphosphohydrolase (ATPDase; CD39)，以及酵素免疫分析法測定培養上清液之TM、von Willebrand factor (vWF)、TF含量。 結果：純化出來含有APL的IgG與HUVEC共同培養後，所有12個病人的IgG及單株抗體IS6造成內皮細胞表面的抗凝固分子－TM明顯降低，3個病人的IgG及單株抗體IS4造成內皮細胞表面的TF增加，所有病人的IgG及單株抗體均未造成內皮細胞表面的CD39改變。1個病人的IgG與HUVEC共同培養後，造成培養上清液的TF增加，所有12個病人的IgG和單株抗體CL1則造成內皮細胞培養上清液的TM減少；4個病人的IgG及所有3株單株抗體造成內皮細胞培養上清液的vWF增加。 結論：從APS病人血清分離出的抗體IgG及單株抗體會影響內皮細胞對血液的凝固，這種抗體作用在內皮細胞，而引起內皮細胞偏向促進凝固的功能，造成血管栓塞的症狀。
並列摘要
Objective. To verify the possible mechanism for thrombosis of Antiphospholipid Syndrome(APS) and the impact of antiphospholipid antibodies(APL) on endothelial cells. Methods. Human umbilical cord vein cells (HUVECs) were incubated with IgG isolated from 12 patients with APS and 3 aCL hybridoma monoclonal antibodies for 6 hours. Expressions of Thrombomodulin (TM), Tissue Factor (TF) and ATP Diphosphosphohydrolase(ATPDase; CD39) were analyzed on cell surface by Flowcytometry. Levels of soluble TM, soluble TF, von Willebrand factor(vWF) were measured in cultural supernatant by ELISA. Results. Incubation with IgG from 12 patients of APS and IS6 aCL hybridoma monoclonal antibody resulted in a significant decrease of TM on HUVECs. Incubation with IgG from 3 APS patients and IS4 aCL hybridoma monoclonal antibody resulted in a significant increase of TF on HUVECs. No significant differences were detected in CD39 on HUVECs while being incubated with IgG from 12 APS patients, 3 aCL hybridoma monoclonal antibody and 3 normal serum. Soluble TF increased in cultural supernatant after treatment with IgG from 1 APS patient. Soluble TM decreased in cultural supernatant after treatment with IgG from 12 APS patients and CL1 hybridoma monoclonal antibody. vWF level in cultural supernatant increased after treatment with IgG from 4 APS patients and 3 hybridoma monoclonal antibodies. Conclusion. These findings suggest that IgG from patients with APS and hybridoma monoclonal antibodies may create a hypercoagulable state by interaction with endothelial cells. The activation of ECs by aPL is an important mechanism that may precede thrombus formation in patients with APS.