细胞黏附分子-1在急性髓性白血病细胞与内皮细胞黏附中的作用

目的　检测急性髓性白血病 (AML)细胞与内皮细胞的黏附及细胞黏附分子 1(ICAM 1)及其配体淋巴细胞功能相关抗原 1(LFA 1)在黏附中的作用。方法　观察AML细胞与静止内皮细胞和肿瘤坏死因子α(TNFα)激活的内皮细胞的黏附 ;AML细胞与内皮细胞混合培养 2 4h的黏附 ;正常中性粒细胞与AML细胞培养上清作用 2 4h后的内皮细胞的黏附 ;流式和ELISA方法检测AML细胞培养上清作用后内皮细胞ICAM 1及可溶性ICAM 1的表达 ;并用ICAM 1和LFA 1的抗体进行阻滞黏附试验。结果　AML细胞与静止的内皮细胞黏附较少 (2 4 33± 2 87) % ,AML细胞与TNFα激活的内皮细胞的黏附 ,与内皮细胞混合培养 2 4h后的黏附以及正常中性粒细胞与AML细胞培养上清作用后内皮细胞的黏附明显增加 ,分别为 (81 87± 4 0 8) % ,(82 0 6± 7 0 5 ) % ,(83 99± 3 86 ) % (n =2 1,P <0 0 0 1) ;静止的内皮细胞ICAM 1及可溶性ICAM 1的表达分别为 (5 5 81± 4 11) %和 (0 839± 0 2 36 )μg/L ;AML细胞培养上清作用后内皮细胞ICAM 1及可溶性ICAM 1的表达明显增加 ,分别为 (6 5 36±5 97) %和 (1 4 2 4± 0 4 6 9) μg/L(n =2 1,P <0 0 5 ) ;用ICAM 1及LFA 1的抗体进行黏附阻滞后 ,AML细胞与TNFα激活的内皮细胞的黏附下降为 (2 0 12±

The role of intercellular adhesion molecule-1 in binding of acute myeloid leukemic blasts cells to human umbilical vein endothelial cells

OBJECTIVE: To evaluate the adhesion of acute myeloid leukemia (AML) blasts to human umbilical vein endothelial cells (HUVECs) and the roles of intercellular adhesion molecule-1 (ICAM-1) and its ligand lymphocyte function associated antigen-1 (LFA-1) in binding of leukemic blasts to HUVECs. METHODS: AML blasts attachment to unactivated or tumor necrosis factor alpha (TNF alpha) activated-endothelial cell monolayers was investigate in vitro; The adhesion of leukemic blasts co cultured with unactivated endothelial cells under static conditions at 37 degrees C for 24 hours was observed;The binding of neutrophils and unactivated endothelial cell monolayers exposed to supernatant of blasts were tested. ICAM-1 on endothelial cell surface and sICAM-1 of endothelial cell supernatant were determined by flow cytometry and ELISA detection. We also observed the adhesion of leukemic blasts in the presence of the adhesion blocking mAbs anti-ICAM-1 and anti-LFA-1. RESULTS: This study has shown that the blast cells attached to unactivated endothelium was little (24.33 +/- 2.87)% and increased after exposure of endothelium to TNF alpha (81.87 +/- 4.08)% (n = 21, P < 0.001); The binding of blasts to endothelium also increased significantly after 24 hours co cultured with unactivated HUVEC (82.06 +/- 7.05)%, (n = 21, P < 0.001); The adhesion of neutrophils and unactivated endothelial cell monolayers exposed to supernatant of blasts was increased significantly (83.99 +/- 3.86)%, (n = 21, P < 0.001). Lower levels of ICAM-1 and sICAM-1 expression was detected on unactivated HUVECs (55.81 +/- 4.11)%, (0.839 +/- 0.236) microg/L respectively. Treatment of HUVECs with AML blasts supernatant for 24 hours increased the expression of ICAM-1 (65.36 +/- 5.97)%, (1.424 +/- 0.469) microg/L respectively (n = 21, P < 0.05) and anti-ICAM-1 and anti-LFA-1 significantly inhibited the adhesion of AML blasts attachment to TNF alpha activated-endothelial cell monolayers (20.12 +/- 1.73)%, (n = 10, P < 0.001). CONCLUSION: Our results indicate that leukemic blasts have the ability to generate some factors that stimulate endothelial cell to secrete ICAM-1 ane to promote their own adhesion to vascular endothelium, interaction of ICAM-1 and its ligand LFA-1 has a key role in adhesion of leukemic blasts and HUVECs.