Shear-induced capping of L-selectin on the neutrophil surface during centrifugation.
L-selectin on leukocytes is critical in leukocyte tethering and adhesion to inflamed endothelium and lymphocyte homing to lymphoid organs. The spatial distribution of L-selectin on leukocytes controls cellular adhesive function in hydrodynamic shear. How L-selectin changes its position on the cell membrane remains an open question, but a possible candidate is shear stress encountered on the cell surface. Here we demonstrate shear-induced L-selectin polarization on the membrane during the process of centrifugation of resting neutrophils via immunofluorescent microscopy. It was found that randomly distributed L-selectin on neutrophils moves to a polar cap at one end of the cell after centrifugation (300 x g for 2 min) without inflammatory stimuli. This L-selectin redistribution under shear was predicted by Monte Carlo simulations that show how convection dominates over diffusion, leading to L-selectin cap formation during centrifugation at 280 x g or during leukocyte adhesion to the endothelial wall at 1 dyn/cm(2). Those results point to a role for shear stress in the modulation of L-selectin distribution, and suggest a possible alternate mechanism and reinterpretation of previous in vitro studies of L-selectin mediated adhesion of neutrophils isolated via centrifugation.
Lee
Dooyoung
D
Department of Chemical Engineering, University of Rochester, Rochester, NY 14627, USA.
King
Michael R
MR
D002448:Cell Adhesion / Q000502:physiology; D006801:Humans; D007152:Immunologic Capping; D019041:L-Selectin / Q000378:metabolism*; D036904:Leukocyte Rolling / Q000502:physiology; D008856:Microscopy, Fluorescence; D009010:Monte Carlo Method; D009504:Neutrophils / Q000378:metabolism*; D013314:Stress, Mechanical
D019041:L-Selectin