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 (0.2×2 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.

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