摘要
目的观察PTA1分子在活化人内皮细胞的表达、调变及其所参与的粘附功能。方法用间接免疫荧光染色结合流式细胞仪分析观察PTA1在内皮细胞的表达和分布 ;用RT PCR方法检测内皮细胞中PTA1mRNA的表达 ;用Na251CrO4 标记静止和TPA活化Jurkat细胞 ,采用4h粘附实验观察不同条件下内皮细胞和Jurkat细胞间的粘附以及PTA1/Ig融合蛋白的粘附阻断作用。结果①静止内皮细胞表达很弱的PTA1分子 ,TNF α、LPS可显著上调PTA1在内皮细胞的表达 ,TNF α刺激8h、LPS刺激2d后PTA1分子在内皮细胞的表达达到高峰。用RT PCR方法在TNF α刺激6h后的内皮细胞中可检测到较高水平的PTA1mRNA ,但静止内皮细胞中只能检测到极微弱的PTA1mRNA。②TNF α活化内皮细胞与Jurkat细胞的粘附作用明显高于静止内皮细胞与Jurkat细胞的粘附作用 ,TPA活化Jurkat细胞与内皮细胞间的粘附作用明显高于静止Jurkat细胞与内皮细胞间的粘附作用 ,且这两种粘附作用均可部分被PTA1/Ig 融合蛋白所阻断。结论TNF α和LPS刺激后的活化内皮细胞可表达较高水平的PTA1mRNA和PTA1蛋白 ,且此PTA1分子可直接参与活化内皮细胞和Jurkat细胞间的粘附 ,表明PTA1是内皮细胞上一种新的可诱导性粘附分子。
Aim To investigate the expression and regulation of inducible PTA1 on the human umbilical vein endothelial cells(HUVEC) and ECV304 cells and its adhesive role in the adhesion between endothelial cells and Jurkat cells. Methods Indirect fluorescent staining, FCM analysis and RT PCR were employed to detect the expression and regulation of PTA1 at the levels of PTA1 protein and PTA1mRNA. Na251CrO4 labeled resting or TPA activated Jurkat cells were added into HUVEC in the presence or absence of PTA1/Ig fusion protein to examine the adhesion effect. Results ①The expressions of PTA1 molecule and PTA1mRNA were very weak on resting HUVEC and ECV304 cells, whereas high level expression could be observed when these cells were stimulated by TNF αor LPS. After 8h stimulation with TNF αor 48h stimulation with LPS the expression of PTA1 protein reached its peak. ②The adhesion between TNF αactivated HUVEC and Jurkat cells was much higher than that between resting HUVEC and Jurkat cells. Similarly the adhesion between TPA activated Jurkat cells and HUVEC was higher than that between resting Jurkat cells and HUVEC. The binding activities between endothelial cells and Jurkat cells either in resting or in activated status could partly be blocked by the addition of PTA1/Ig fusion protein. Conclusion TNF αor LPS activated HUVEC and ECV304 cells can express high level PTA1 protein and PTA1mRNA,and PTA1 is involved in the adhesion between endothelial and Jurkat cells,suggesting that PTA1 is a novel inducible adhesion molecule on human endothelial cells.
出处
《细胞与分子免疫学杂志》
CSCD
2000年第1期23-26,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金!No.39770697
