Klein C L, Bittinger F, Köhler H, Wagner M, Otto M, Hermanns I, Kirkpatrick C J
Institute of Pathology, Johannes Gutenberg University, Mainz, Germany.
Pathobiology. 1995;63(2):83-92. doi: 10.1159/000163938.
Endothelial cells (EC) are very responsive to proinflammatory cytokines, e.g. interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF alpha), as well as to bacterial lipopolysaccharide. EC are stimulated by these substances to secrete chemotactic factors and to increase expression of cell adhesion molecules (CAM), leading to dramatically altered interactions with leukocytes, e.g. granulocytes and monocytes. In these interactions E-selectin, ICAM-1 and VCAM-1 are known to play an important role, as they are presented by the EC and interact with corresponding ligands on the white blood cell membranes. These adhesion molecules have been studied worldwide in a variety of in vitro experiments using cultured EC. Different passages and mixtures of passages have been used in these experiments, often without any regard to the comparability of the results. In this study the expression of E-selectin, ICAM-1 and VCAM-1 on cultured human umbilical vein EC (HUVEC) obtained from different passages (passages 1-6) was studied after 4, 8 and 24 h of exposure to IL-1 beta and TNF alpha. In previous studies, we have shown that IL-1 beta and TNF alpha increase the expression of E-selectin and ICAM-1 on the cytoplasmatic membranes of HUVEC and human adult EC from the saphenous vein and femoral artery in a similar fashion. Using a comparative quantitative cell enzyme immunoassay, we found that the expression of the adhesion molecules was significantly reduced with increasing passages. There was also a decreased persistence of CAM comparing different periods of stimulation between 6 and 24 h in the different passages. These data indicate that the number of passages plays an important role in the expression of adhesion molecules on EC. The results are relevant for the meaningful planning of comparative in vitro studies on EC presentation of CAM.
内皮细胞（EC）对促炎细胞因子，如白细胞介素 -1（IL -1）和肿瘤坏死因子α（TNFα）以及细菌脂多糖非常敏感。这些物质刺激内皮细胞分泌趋化因子并增加细胞黏附分子（CAM）的表达，从而导致与白细胞，如粒细胞和单核细胞的相互作用发生显著改变。在这些相互作用中，已知E -选择素、细胞间黏附分子 -1（ICAM -1）和血管细胞黏附分子 -1（VCAM -1）起着重要作用，因为它们由内皮细胞呈现并与白细胞膜上的相应配体相互作用。在世界各地，人们使用培养的内皮细胞在各种体外实验中对这些黏附分子进行了研究。在这些实验中使用了不同的传代次数和传代混合物，结果往往没有考虑到结果的可比性。在本研究中，研究了从不同传代次数（第1 - 6代）获得的培养人脐静脉内皮细胞（HUVEC）在暴露于IL -1β和TNFα 4、8和24小时后E -选择素、ICAM -1和VCAM -1的表达。在先前的研究中，我们已经表明，IL -1β和TNFα以类似的方式增加HUVEC以及来自大隐静脉和股动脉的成人内皮细胞细胞质膜上E -选择素和ICAM -1的表达。使用比较定量细胞酶免疫测定法，我们发现随着传代次数的增加，黏附分子的表达显著降低。在不同传代次数中，比较6至24小时不同刺激时间段时，细胞黏附分子的持续性也有所降低。这些数据表明传代次数在内皮细胞黏附分子的表达中起着重要作用。这些结果对于有意义地规划关于内皮细胞呈现细胞黏附分子的比较体外研究具有重要意义。
