Validation of a technique to measure leukocyte adhesion to arterial segments: effects of drug treatments

Adhesion and transmigration of leukocytes into arterial walls occurs after vascular injury and may play a role in the development of atherosclerosis a nd restenosis. This protocol presents a simple, rapid method for quantifyin g leukocyte adhesion to artery segments ex vivo. The procedure involves iso lating leukocytes from rabbit whole blood and labelling with the gamma-emit ting isotope Cr-51. Labelled leukocytes are added to open rings of subclavi an artery taken from the same rabbit. After gamma counting, percentage leuk ocyte adhesion can be calculated with reference to a sample containing a qu antity of labelled leukocytes equivalent to that which was added to the art ery, Leukocyte adhesion was increased by L-NAME, thrombin and increasing in cubation time and decreased by low temperatures. In addition, leukocyte adh esion was found to be increased following a vascular stretch injury perform ed in vitro. This protocol offers a number of advantages: the rapidity of t he leukocyte isolation and labelling; the small quantity of leukocytes requ ired; the ability to use autologous leukocytes; the applicability to whole arteries and arteries injured in vitro or in vivo, allowing the effects of vascular injury on leukocyte adhesion to be studied. (C) 2001 Elsevier Scie nce B.V. All rights reserved.