Gerritsen M E, Kelley K A, Ligon G, Perry C A, Shen C P, Szczepanski A, Carley W W
Institute for Inflammation and Autoimmunity, Miles Inc., West Haven, CT 06516.
Arthritis Rheum. 1993 May;36(5):593-602. doi: 10.1002/art.1780360504.
To examine the regulation of intercellular adhesion molecule 1 (ICAM-1) in human synovial microvascular endothelial cells (HSE) and human umbilical vein endothelial cells (HUVE) upon exposure to a variety of agents.
Cultured endothelial cells were treated with various cytokines alone and in combination. The expression of ICAM-1 was evaluated at several levels, including an investigation of messenger RNA (mRNA) and surface protein expression.
Treatment of HSE with interleukin-1 alpha (IL-1 alpha) or tumor necrosis factor alpha (TNF alpha) resulted in minimal increases in ICAM-1 expression, in contrast to findings with HUVE. Incubation of HUVE or HSE with IL-1 or TNF in combination with interferon-gamma (IFN gamma) greatly potentiated the increase in ICAM-1 surface expression. The synergistic effect of IFN gamma and TNF was confirmed by several methods, including a cell-based enzyme-linked immunosorbent assay, fluorescence-activated cell sorting, immunofluorescence staining, and determination of mRNA levels. IFN gamma also augmented the actions of several other agonists on HSE, i.e., IL-4, lipopolysaccharide, and TNF beta/lymphotoxin. Immunoprecipitation of TNF alpha + IFN gamma-stimulated, 125I-labeled HSE cells with anti-ICAM-1 revealed a single 90-kd band, similar in size to ICAM-1 from HUVE treated in an identical manner. Unexpectedly, IFN gamma alone was a potent stimulus for HSE ICAM-1 mRNA synthesis, but was relatively ineffective in HUVE.
These studies indicate that IFN gamma plays a critical synergistic role in the regulation of ICAM-1 expression in human synovial endothelial cells.
研究人滑膜微血管内皮细胞（HSE）和人脐静脉内皮细胞（HUVE）在暴露于多种试剂时细胞间黏附分子1（ICAM-1）的调控情况。
将培养的内皮细胞单独或联合用多种细胞因子处理。在几个水平评估ICAM-1的表达，包括信使核糖核酸（mRNA）和表面蛋白表达的研究。
与HUVE的研究结果相反，用白细胞介素-1α（IL-1α）或肿瘤坏死因子α（TNFα）处理HSE导致ICAM-1表达仅有极小增加。将HUVE或HSE与IL-1或TNF联合干扰素-γ（IFNγ）孵育极大地增强了ICAM-1表面表达的增加。通过几种方法证实了IFNγ和TNF的协同作用，包括基于细胞的酶联免疫吸附测定、荧光激活细胞分选、免疫荧光染色和mRNA水平测定。IFNγ还增强了其他几种激动剂对HSE的作用，即IL-4、脂多糖和TNFβ/淋巴毒素。用抗ICAM-1对TNFα + IFNγ刺激的、125I标记的HSE细胞进行免疫沉淀，显示出一条单一的90-kd条带，其大小与以相同方式处理的HUVE的ICAM-1相似。出乎意料的是，单独的IFNγ是HSE中ICAM-1 mRNA合成的有效刺激物，但在HUVE中相对无效。
这些研究表明IFNγ在人滑膜内皮细胞中ICAM-1表达的调控中起关键的协同作用。
