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 and restenosis. This protocol presents a simple, rapid method for quantifying leukocyte adhesion to artery segments ex vivo. The procedure involves isolating leukocytes from rabbit whole blood and labelling with the gamma-emitting isotope 51Cr. Labelled leukocytes are added to open rings of subclavian artery taken from the same rabbit. After gamma counting, percentage leukocyte adhesion can be calculated with reference to a sample containing a quantity of labelled leukocytes equivalent to that which was added to the artery. Leukocyte adhesion was increased by L-NAME, thrombin and increasing incubation time and decreased by low temperatures. In addition, leukocyte adhesion was found to be increased following a vascular stretch injury performed in vitro. This protocol offers a number of advantages: the rapidity of the leukocyte isolation and labelling; the small quantity of leukocytes required; 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.
Kennedy
S
S
Department of Physiology and Pharmacology, Strathclyde Institute of Biological Sciences, University of Strathclyde, 27 Taylor Street, G4 0NR, Scotland, Glasgow, UK.
Miller
A M
AM
Wadsworth
R M
RM
McPhaden
A R
AR
Wainwright
C L
CL
D000818:Animals; D001158:Arteries / Q000187:drug effects / Q000293:injuries* / Q000473:pathology; D002404:Catheterization / Q000009:adverse effects; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology*; D066298:In Vitro Techniques; D007962:Leukocytes / Q000187:drug effects* / Q000502:physiology*; D008297:Male; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D011817:Rabbits; D013314:Stress, Mechanical; D013696:Temperature; D013917:Thrombin / Q000187:drug effects; D013997:Time Factors
D013917:Thrombin; D019331:NG-Nitroarginine Methyl Ester