Shih P T, Elices M J, Fang Z T, Ugarova T P, Strahl D, Territo M C, Frank J S, Kovach N L, Cabanas C, Berliner J A, Vora D K
Department of Pathology, University of California-Los Angeles, California 90095-1732, USA.
J Clin Invest. 1999 Mar;103(5):613-25. doi: 10.1172/JCI5710.
We have shown previously that treatment of human aortic endothelial cells (HAECs) with minimally modified low-density lipoprotein (MM-LDL) induces monocyte but not neutrophil binding. This monocyte binding was not mediated by endothelial E-selectin, P-selectin, vascular cell adhesion molecule-I, or intercellular adhesion molecule-I, suggesting an alternative monocyte-specific adhesion molecule. We now show that moncytic alpha4beta1 integrins mediate binding to MM-LDL-treated endothelial cells. We present data suggesting that the expression of the connecting segment-1 (CS-1) domain of fibronectin (FN) is induced on the apical surface of HAEC by MM-LDL and is the endothelial alpha4beta1 ligand in MM-LDL-treated cells. Although the levels of CS-1 mRNA and protein were not increased, we show that MM-LDL treatment causes deposition of FN on the apical surface by activation of beta1integrins, particularly those associated with alpha5 integrins. Activation of beta1 by antibody 8A2 also induced CS-1-mediated monocyte binding. Confocal microscopy demonstrated the activated beta1 and CS-1colocalize in concentrated filamentous patches on the apical surface of HAEC. Both anti-CS-1 and an antibody to activated beta1 showed increased staining on the luminal endothelium of human coronary lesions with active monocyte entry. These results suggest the importance of these integrin ligand interactions in human atherosclerosis.
我们之前已经表明,用轻度修饰的低密度脂蛋白(MM-LDL)处理人主动脉内皮细胞(HAECs)会诱导单核细胞而非中性粒细胞的黏附。这种单核细胞黏附不是由内皮细胞E-选择素、P-选择素、血管细胞黏附分子-1或细胞间黏附分子-1介导的,提示存在一种替代性的单核细胞特异性黏附分子。我们现在表明,单核细胞的α4β1整合素介导与MM-LDL处理的内皮细胞的黏附。我们提供的数据表明,纤连蛋白(FN)连接段-1(CS-1)结构域的表达在MM-LDL作用下于HAEC的顶端表面被诱导,并且是MM-LDL处理细胞中内皮细胞α4β1的配体。尽管CS-1 mRNA和蛋白水平没有增加,但我们表明MM-LDL处理通过激活β1整合素,特别是与α5整合素相关的β1整合素,导致FN在顶端表面沉积。抗体8A2对β1的激活也诱导了CS-1介导的单核细胞黏附。共聚焦显微镜显示活化的β1和CS-1共定位于HAEC顶端表面的浓缩丝状斑块中。抗CS-1抗体和活化β1的抗体在有活跃单核细胞进入的人冠状动脉病变的管腔内皮层上均显示染色增加。这些结果表明这些整合素配体相互作用在人类动脉粥样硬化中的重要性。