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Synthesis and Surface Expression of CD14 by Human Endothelial Cells

机译：人内皮细胞合成CD14及其表面表达

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Previous studies have reported that human vascular endothelial cells lack the membrane-bound lipopolysaccharide (LPS) receptor, CD14 (mCD14). By optimizing assay conditions, including the selection of anti-CD14 monoclonal antibody, we now demonstrate that human umbilical vein endothelial cells (HUVEC) express CD14 on the cell surface. Single-passage HUVEC showed approximately 20 times less expression of CD14 than monocytes. Interestingly, there was significant loss of surface CD14 expression with increasing numbers of culture passages. Evidence for synthesis of CD14 by HUVEC was provided by the finding that l-[35S]methionine was incorporated into CD14. In addition, the expression of CD14 on HUVEC was upregulated by LPS, lysophosphatidic acid, and tissue culture supplements, and this upregulation was dependent on protein synthesis. Furthermore, the results imply that mCD14 is required for LPS-induced activation of endothelial cells in the absence of serum and that it acts in concert with serum factors (soluble CD14). Our results provide evidence that CD14 is expressed by endothelial cells and suggest that the previous inability to observe expression of this molecule has been due to culture and staining conditions. This finding has important implications for the understanding of the mechanisms by which LPS stimulates endothelial cells and the management of sepsis caused by gram-negative bacteria.
机译：先前的研究已报道人血管内皮细胞缺乏膜结合脂多糖（LPS）受体CD14（mCD14）。通过优化分析条件，包括选择抗CD14单克隆抗体，我们现在证明人脐静脉内皮细胞（HUVEC）在细胞表面表达CD14。单代HUVEC显示CD14的表达比单核细胞少约20倍。有趣的是，随着培养传代次数的增加，表面CD14表达明显丧失。 HUVEC通过将1-[ 35 S]蛋氨酸掺入CD14的发现，提供了HUVEC合成CD14的证据。此外，LPS，溶血磷脂酸和组织培养补剂会上调HUVEC上CD14的表达，而这种上调取决于蛋白质的合成。此外，结果暗示在没有血清的情况下，mCD14是LPS诱导的内皮细胞活化所必需的，并且它与血清因子（可溶性CD14）协同作用。我们的结果提供了内皮细胞表达CD14的证据，并表明以前无法观察到该分子的表达是由于培养和染色条件所致。这一发现对于理解LPS刺激内皮细胞的机制以及由革兰氏阴性细菌引起的败血症的处理具有重要意义。

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