© 2001 American Heart Association, Inc.
Vascular Biology |
Cross-Linking of Brain Endothelial Intercellular Adhesion Molecule (ICAM)-1 Induces Association of ICAM-1 With Detergent-Insoluble Cytoskeletal Fraction
From the Division of Endothelial and Epithelial Cell Biology, Institute of Ophthalmology, University College London, London, UK, and the Centre for Cardiovascular Biology and Medicine (J.D.P.), Kings College London, London, UK.
Correspondence to Dr Peter Adamson, Division of Endothelial and Epithelial Cell Biology, Institute of Ophthalmology, University College London 11-43 Bath Street, London, UK EC1V 9EL. E-mail padamson{at}hgmp.mrc.ac.uk
Abstract—Intercellular adhesion molecule (ICAM)-1 plays a vital role in the process of leukocyte transmigration through endothelial cell (EC) barriers and has been shown to mediate signal transduction events in ECs induced either by its cross-linking or by the binding of T lymphocytes. Immunoblotting of ICAM-1 of Triton X-100 detergent fractions demonstrated that the majority of ICAM-1 was contained within the detergent-soluble fraction (noncytoskeletal associated) under basal conditions. After cross-linking of endothelial ICAM-1 with monoclonal antibody or coculture with T lymphocytes, EC ICAM-1 was observed to partition with a Triton X-100–insoluble (cytoskeletal associated) fraction in a dose- and time-dependent manner. Redistribution of ICAM-1 was specific, inasmuch as no association with the Triton X-100–insoluble fraction was observed after cross-linking of vascular cell adhesion molecule-1, nor did cross-linking of ICAM-1 result in a redistribution of the platelet and EC adhesion molecule. ICAM-1 association with the endothelial cytoskeleton after cross-linking was unaffected after treatment of the cells with cytochalasin D, C3-transferase, removal of extracellular calcium ions, or chelation of intracellular calcium ions. These data show that ICAM-1 colocalizes with the endothelial cytoskeleton and associates with a detergent-insoluble fraction after cross-linking.
Key Words: endothelium • intercellular adhesion molecule-1 • cytoskeleton
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