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Immunoenzymometric analysis for expression and shedding of intercellular adhesion molecule-1 on human endothelial cells stimulated with cytokines or lipopolysaccharide

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Abstract

Unstimulated endothelial cell (EC)cultures express low levels of intercellular adhesion molecule-1 (ICAM-1) and their expression can be enhanced by inflammatory cytokines such as tumor necrosis factor α (TNF). Three monoclonal antibodies (MoAbs) highly reactive with TNF-stimulated human ECs were established and defined to recognize a 95 kDa cell surface protein specifically expressed on cytokine-activated ECs, which was immunochemically identified as ICAM-1. The quantitative immunoassay of soluble and insoluble ICAM-1 could be performed with two different MoAbs. Secretion of fibronectin or the von Willebrand factor, was not significantly enhanced with TNF stimulation. Cellular expression of ICAM-1 was drastically induced by TNF or interleukin-1 stimulation, and the moderate expression with delayed-action was observed only by lipopolysaccharide stimulation. A maximal amount of soluble ICAM-1 was released from ECs stimulated only by TNF, apparently in a dose dependent manner, but no significant release of ICAM-1 was induced by thrombin interleukin-2, or lipopolysacchardes. Released levels of soluble ICAM-1 from interleukin-1-stimulated ECs were apparently diminished as compared with those from TNF-stimulated cells. These results suggest that release of soluble ICAM-1 from EC surfaces can be most significantly enhanced by TNF-specific signaling, and prospectively, should be a sensitive indicator of intravascular inflammation in acute endothelium injury.

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Authors and Affiliations

  1. Cell Technology Reagent Section, Biotechnology Research Laboratories, Takara Shuzo Company Limited, Seta 3-4-1, 520-21, Otsu, Shiga, Japan

    Yuka Sano, Sayuri Hirai, Masahiko Katayama & Ikunoshin Kato

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  1. Yuka Sano
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  2. Sayuri Hirai
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  3. Masahiko Katayama
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  4. Ikunoshin Kato
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Sano, Y., Hirai, S., Katayama, M. et al. Immunoenzymometric analysis for expression and shedding of intercellular adhesion molecule-1 on human endothelial cells stimulated with cytokines or lipopolysaccharide. Mol Cell Biochem 139, 123–130 (1994). https://doi.org/10.1007/BF01081735

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  • DOI: https://doi.org/10.1007/BF01081735

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