Burke-Gaffney A, Keenan A K
Children's Research Centre, Our Lady's Hospital for Sick Children, Crumlin, Dublin, Ireland.
Immunopharmacology. 1993 Jan-Feb;25(1):1-9. doi: 10.1016/0162-3109(93)90025-l.
The permeability of human umbilical vein endothelial cell (HUVEC) monolayers to [125I]-labelled bovine serum albumin (BSA) was examined following pretreatment of the cells with various cytokines. The electrical resistance measured across untreated, confluent, intact HUVEC monolayers was 18.2 +/- 3.8 omega.cm2 (mean +/- S.D. of 4 observations). Human recombinant (hr) interleukin-1 alpha/beta (IL-1 alpha/beta), hr tumor necrosis factor-alpha (TNF-alpha), and hr interferon-gamma (IFN-gamma) each increased HUVEC monolayer permeability in a time- and dose-dependent manner. These effects were inhibitable by neutralizing antibodies (nAb) to the corresponding cytokines, and were not due to contamination by endotoxin (abolition of cytokine effect by heat treatment, and no effect on cytokine action of the endotoxin inhibitor polymyxin B). The effects of these cytokines were not due to endothelial cell (EC) interleukin-6 (IL-6) induction (IL-6 shown not to increase permeability) and the effect of hrTNF-alpha could not be accounted for by induction of IL-1 (effect not inhibited by hrIL-1 alpha/beta nAb). The effects of three different combinations of the cytokines (each combination at two different concentrations) on HUVEC monolayer permeability were also examined. hrIFN-gamma with hrTNF-alpha or hrIL-1 alpha/beta gave an increase in permeability (at both concentration combinations) greater than that seen with either cytokine alone. hrTNF-alpha and hrIL-1 alpha/beta in combination however produced an enhanced effect only at low concentrations, high concentrations in combination producing an effect no greater than either agent alone. These results highlight the importance of investigating actions of cytokine combinations on in vitro models of endothelial cell activation.
在用各种细胞因子对人脐静脉内皮细胞（HUVEC）单层进行预处理后，检测了其对[125I]标记的牛血清白蛋白（BSA）的通透性。在未处理的、汇合的、完整的HUVEC单层上测得的电阻为18.2±3.8Ω·cm2（4次观察的平均值±标准差）。人重组（hr）白细胞介素-1α/β（IL-1α/β）、hr肿瘤坏死因子-α（TNF-α）和hr干扰素-γ（IFN-γ）均以时间和剂量依赖性方式增加HUVEC单层的通透性。这些作用可被针对相应细胞因子的中和抗体（nAb）抑制，且不是由内毒素污染所致（热处理消除细胞因子作用，内毒素抑制剂多粘菌素B对细胞因子作用无影响）。这些细胞因子的作用不是由于内皮细胞（EC）白细胞介素-6（IL-6）的诱导（IL-6未显示增加通透性），且hrTNF-α的作用不能用IL-1的诱导来解释（作用不被hrIL-1α/β nAb抑制）。还检测了细胞因子的三种不同组合（每种组合有两种不同浓度）对HUVEC单层通透性的影响。hrIFN-γ与hrTNF-α或hrIL-1α/β组合时通透性增加（在两种浓度组合下），大于单独使用任何一种细胞因子时的增加幅度。然而，hrTNF-α和hrIL-1α/β组合仅在低浓度时产生增强作用，高浓度组合时产生的作用不大于单独使用任何一种药物。这些结果突出了研究细胞因子组合对内皮细胞激活体外模型作用的重要性。
