Open AccessRegulation of the expression of intercellular adhesion molecule 1 in cultured human endothelial cells derived from rheumatoid synovium
Author(s) -
Gerritsen Mary E.,
Kelley Keith A.,
Ligon Gwenda,
Perry Carol A.,
Shen ChienPing,
Szczepanski Andrew,
Carley William W.
Publication year - 1993
Publication title -
arthritis & rheumatism
Language(s) - English
Resource type - Journals
eISSN - 1529-0131
pISSN - 0004-3591
DOI - 10.1002/art.1780360504
Subject(s) - tumor necrosis factor alpha , umbilical vein , microbiology and biotechnology , biology , intercellular adhesion molecule 1 , cell sorting , icam 1 , cytokine , endothelial stem cell , interferon gamma , cell adhesion molecule , lipopolysaccharide , intracellular , immunology , in vitro , flow cytometry , biochemistry
Objective. 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. Methods. 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. Results. Treatment of HSE with interleukin‐1α (IL‐1α) or tumor necrosis factor α (TNFα) 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‐γ (IFNγ) greatly potentiated the increase in ICAM‐1 surface expression. The synergistic effect of IFNγ 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. IFNγ also augmented the actions of several other agonists on HSE, i.e., IL‐4, lipopolysaccharide, and TNFβ/lymphotoxin. Immunoprecipitation of TNFα + IFNγ–stimulated, 125 I‐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, IFNγ alone was a potent stimulus for HSE ICAM‐1 mRNA synthesis, but was relatively ineffective in HUVE. Conclusion. These studies indicate that IFNγ plays a critical synergistic role in the regulation of ICAM‐1 expression in human synovial endothelial cells.