Autoperfused mouse flow chamber reveals synergistic neutrophil accumulation through P-selectin and E-selectin.
To study rolling of mouse neutrophils on P- and E-selectins in whole blood and without cell isolation, we constructed an autoperfused flow chamber made from rectangular microslides (0.2x2 mm) perfused from a carotid artery catheter. A differential pressure transducer served to measure wall shear stress. Green fluorescent neutrophils rolled on P-selectin but not E-selectin coated at 50 ng/ml, with some rolling on E-selectin at 150 ng/ml. However, when P- and E-selectins were coimmobilized, the resulting number of rolling neutrophils was sixfold and fourfold higher than on P- or E-selectin alone. Velocity and flux analysis shows that P-selectin initiates neutrophil rolling, and a small amount of E-selectin, unable to capture many neutrophils, reduces the rolling velocity of all neutrophils by more than 90%. The unexpected synergism between E- and P-selectins explains why neutrophil recruitment is enhanced when both selectins are expressed.
Smith
Michael L
ML
Department of Biomedical Engineering, University of Virginia, Charlottesville, USA.
Sperandio
Markus
M
Galkina
Elena V
EV
Ley
Klaus
K
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D015194:Diffusion Chambers, Culture / Q000295:instrumentation / Q000379:methods; D004357:Drug Synergism; D019040:E-Selectin / Q000378:metabolism* / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D036904:Leukocyte Rolling / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism* / Q000494:pharmacology; D013314:Stress, Mechanical; D014160:Transducers, Pressure
D019040:E-Selectin; D019007:P-Selectin; D049452:Green Fluorescent Proteins