Amberger A, Maczek C, Jürgens G, Michaelis D, Schett G, Trieb K, Eberl T, Jindal S, Xu Q, Wick G
Institute for Biomedical Aging Research, Austrian Academy of Sciences, Innsbruck, Austria.
Cell Stress Chaperones. 1997 Jun;2(2):94-103. doi: 10.1379/1466-1268(1997)002<0094:ceoive>2.3.co;2.
T-cells and monocytes are the first cells infiltrating the arterial intima during the early stages of atherogenesis. Recently our laboratory has provided evidence that T-cells isolated from atherosclerotic intima reacts against heat shock protein 60 (Hsp60). Transmigration of activated T-cells into the intima is mediated by adhesion molecules (ICAM-1; VCAM-1; ELAM-1) expressed on activated endothelial cells. Here we studied the potential of cytokines (TNF-alpha, IFN-gamma, IL-1). Escherichia coli lipopolysaccharide (LPS), native and oxidized low-density lipoprotein (LDL; oxLDL) and high temperature to induce adhesion molecules as well as Hsp60 and Hsp70 expression in human endothelial cells (EC). On Northern blots, a strong signal for ICAM-1, VCAM-1 and ELAM-1 was detected after 4 h, which thereafter declined, but did not reach the basal level of untreated control cells. Heat shock induced the expression of Hsp60 and Hsp70 but not of adhesion molecules. EC were cultivated in serum-free medium, which led to the expression of adhesion molecule transcripts. Addition of LDL or oxLDL to these ECs did not alter the expression of these transcripts. The production of adhesion molecule proteins was analysed by flow cytometry. In human venous endothelial cells (HVEC) and human arterial endothelial cells (HAEC) ICAM-1 and VCAM-1 production was permanently highly induced, whereas the high level of ELAM-1 production at 4 h disappeared after 24 h. Furthermore, only HAEC, but not HVEC, produced ICAM-1, VCAM-1 and ELAM-1 after stress by moderately and highly oxLDL. LDL and oxLDL did not induce the production of Hsp60 and Hsp70. The present study demonstrates the co-expression of Hsp60 and adhesion molecules in arterial and venous EC in response to cytokine and LPS exposure, and that oxLDL is an efficient inducer of adhesion molecules in arterial EC and not in venous EC. These features provide the prerequisites for a cellular immune reaction against Hsp60 expressed by stressed EC in the initial stages of atherosclerosis.
在动脉粥样硬化形成的早期阶段，T细胞和单核细胞是最早浸润动脉内膜的细胞。最近我们实验室提供的证据表明，从动脉粥样硬化内膜分离出的T细胞会对热休克蛋白60（Hsp60）产生反应。活化的T细胞向内膜的迁移是由活化内皮细胞上表达的黏附分子（ICAM-1、VCAM-1、ELAM-1）介导的。在此，我们研究了细胞因子（肿瘤坏死因子-α、干扰素-γ、白细胞介素-1）、大肠杆菌脂多糖（LPS）、天然和氧化型低密度脂蛋白（LDL；oxLDL）以及高温在人内皮细胞（EC）中诱导黏附分子以及Hsp60和Hsp70表达的潜力。在Northern印迹上，4小时后检测到ICAM-1、VCAM-1和ELAM-1的强信号，此后信号减弱，但未达到未处理对照细胞的基础水平。热休克诱导Hsp60和Hsp70的表达，但不诱导黏附分子的表达。内皮细胞在无血清培养基中培养，这导致黏附分子转录本的表达。向这些内皮细胞中添加LDL或oxLDL不会改变这些转录本的表达。通过流式细胞术分析黏附分子蛋白的产生。在人静脉内皮细胞（HVEC）和人动脉内皮细胞（HAEC）中，ICAM-1和VCAM-1的产生持续被高度诱导，而4小时时ELAM-1的高水平产生在24小时后消失。此外，只有HAEC，而不是HVEC，在受到中度和高度氧化型低密度脂蛋白应激后产生ICAM-1、VCAM-1和ELAM-1。LDL和oxLDL不会诱导Hsp60和Hsp70的产生。本研究表明，在暴露于细胞因子和LPS时，动脉和静脉内皮细胞中Hsp60和黏附分子共同表达，并且氧化型低密度脂蛋白是动脉内皮细胞而非静脉内皮细胞中黏附分子的有效诱导剂。这些特征为动脉粥样硬化初始阶段针对应激内皮细胞表达的Hsp60的细胞免疫反应提供了前提条件。
