HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 286/1/H359    most recent 00491.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via ISI Web of Science (32) Citing Articles via Google Scholar Google Scholar Articles by Zhang, X. Articles by Goligorsky, M. S. Search for Related Content PubMed PubMed Citation Articles by Zhang, X. Articles by Goligorsky, M. S.

Atomic force microscopy measurement of leukocyte-endothelial interaction

¹Department of Physiology and Biophysics, University of Miami School of Medicine, Miami, Florida 33101; ²Department of Medicine, The University of Tokyo, Tokyo 113-8655, Japan; ³Department of Pediatrics, State University of New York, Stony Brook 11794; and ⁴Departments of Medicine and Pharmacology, New York Medical College, Valhalla, New York 10595

Submitted 28 May 2003 ; accepted in final form 10 September 2003

Leukocyte adhesion to vascular endothelium is a key initiating step in the pathogenesis of many inflammatory diseases. In this study, we present real-time force measurements of the interaction between monocytic human promyelocytic leukemia cells (HL-60) cells and a monolayer of human umbilical vein endothelial cells (HUVECs) by using atomic force microscopy (AFM). The detachment of HL-60-HUVEC conjugates involved a series of rupture events with force transitions of 40–100 pN. The integrated force of these rupture events provided a quantitative measure of the adhesion strength on a whole cell level. The AFM measurements revealed that HL-60 adhesion is heightened in the borders formed by adjacent HUVECs. The average force and mechanical work required to detach a single HL-60 from the borders of a tumor necrosis factor--activated HUVEC layer were twice as high as those of the HUVEC bodies. HL-60 adhesion to the monolayer was significantly reduced by a monoclonal antibody against ₁-integrins and partially inhibited by antibodies against selectins ICAM-1 and VCAM-1 but was not affected by anti-_V3. Interestingly, adhesion was also inhibited in a dose-dependent manner (IC₅₀ 100 nM) by a cyclic arginine-glycine-aspartic acid (cRGD) peptide. This effect was mediated via interfering with the VLA-4-VCAM-1 binding. In parallel measurements, transmigration of HL-60 cells across a confluent HUVEC monolayer was inhibited by the cRGD peptide and by both anti-₁ and anti-_V3 antibodies. In conclusion, these data demonstrate the role played by ₁-integrins in leukocyte-endothelial adhesion and transmigration and the role played by _V3 in transmigration, thus underscoring the high efficacy of cRGD peptide in blocking both the adhesion and transmigration of monocytes.

integrins; cell-cell adhesion; arginine-glycine-aspartic acid peptide