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American Journal of Pathology, Vol 147, 1278-1288, Copyright © 1995 by American Society for Investigative Pathology

REGULAR ARTICLES

Apical topography and modulation of ICAM-1 expression on activated endothelium

A Almenar-Queralt, A Duperray, LA Miles, J Felez and DC Altieri
Department of Cell Biology, Scripps Research Institute, La Jolla, California, USA.

Leukocyte-endothelium interactions and general inflammatory responses are contributed by the regulated expression of intercellular adhesion molecule-1 (ICAM-1) on endothelium. It is now shown by confocal fluorescence microscopy and immunogold transmission electron microscopy that ICAM-1 was exclusively localized on the apical (luminal) membrane of cytokine-activated human umbilical vein endothelial cells. In contrast, other cell adhesion-promoting molecules, including beta 1 integrins, were expressed exclusively on the basolateral endothelial cell membrane, under the same experimental conditions. Kinetic binding studies of a 125I-labeled monoclonal antibody to ICAM-1 revealed that approximately 8% of membrane ICAM-1 on cytokine-activated endothelium was internalized in both coated and non-coated vesicles at 37 degrees C, with a t1/2 of approximately 18 min and a rate of approximately 3200 molecules/minute. This internalization pathway was directly dependent upon the level of ICAM-1 expression on the cell surface. Genetically engineered ICAM-1 transfectants, expressing a 10-fold higher receptor density than activated endothelium, internalized approximately 18% of membrane ICAM-1 at a rate of 75,000 molecules/minute with a t1/2 of approximately 22 min. These findings suggest that a combined pathway of polarized membrane topography and receptor trafficking may regulate ICAM-1-dependent adhesion at the site of vascular injury and endothelial cell activation.

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