507f Formation and Deposition of Leukocyte Microparticles during Rolling

Phillip Coghill, Alexander Christov, and David W. Schmidtke. School of Chemical, Biological, and Materials Engineering, University of Oklahoma, 100 East Boyd, Norman, OK 73019

During inflammation and thrombosis, interactions of selectins with cell-surface adhesion molecules allow the tethering and rolling of leukocytes on blood vessel walls. Leukocytes express L-selectin, whereas activated endothelial cells express P- and E-selectin. Leukocyte rolling allows regional sampling of chemokines and other mediators of leukocyte targeting, which leads to integrin-dependent arrest and emigration of leukocytes into the underlying tissues. Rolling requires the rapid formation and rapid dissociation of selectin-ligand bonds that are subjected to tensile forces applied by wall shear stress. We have previously demonstrated the rapid formation of membrane tethers as neutrophils roll on P-selectin. Normally these membrane tethers retract back to the cell body, however we have also observed that membrane tethers can break and deposit membrane microparticle (< 1 &mum) or microfragment (> 1 &mum) on the underlying surface. Breakage of membrane tethers was observed when neutrophils rolled over a wide range of P-selectin site densities (170-1096 sites/&mum2) and shear rates (100-800s-1). We demonstrate that the frequency of membrane tether formation (40-100%) and microparticle deposition (0-40%) both increase as the wall shear stress increases, while there was a small effect of ligand density. We also demonstrate that deposited membrane microfragments may serve as adhesion points for other cells.