Atomic force microscopy measurement of leukocyte-endothelial interaction

• Published in: American journal of physiology. Heart and circulatory physiology 2004, Vol.55 (1), p.H359-H367
• Main Authors: XIAOHUI ZHANG, AILEEN CHEN, DINA DE LEON, HONG LI, NOIRI, Eisei, MOY, Vincent T, GOLIGORSKY, Michael S
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Blood vessels; Cell adhesion & migration; Fundamental and applied biological sciences. Psychology; Immune system; Leukemia; Microscopy; Peptides; Vertebrates: cardiovascular system
• ISSN: 0363-6135; 1522-1539

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-{alpha}-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 {beta}1-integrins and partially inhibited by antibodies against selectins ICAM-1 and VCAM-1 but was not affected by anti-{alpha}V{beta}3. Interestingly, adhesion was also inhibited in a dose-dependent manner (IC50 {approx} 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-{beta}1 and anti-{alpha}V{beta}3 antibodies. In conclusion, these data demonstrate the role played by {beta}1-integrins in leukocyte-endothelial adhesion and transmigration and the role played by {alpha}V{beta}3 in transmigration, thus underscoring the high efficacy of cRGD peptide in blocking both the adhesion and transmigration of monocytes. [PUBLICATION ABSTRACT]