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Published online before print April 1, 2004

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(Journal of Leukocyte Biology. 2004;76:985-993.)
© 2004 by Society for Leukocyte Biology

Autoperfused mouse flow chamber reveals synergistic neutrophil accumulation through P-selectin and E-selectin

Michael L. Smith^*, Markus Sperandio^*,,1, Elena V. Galkina and Klaus Ley^*,

^* Department of Biomedical Engineering and
Cardiovascular Research Center, University of Virginia, Charlottesville; and
Neonatal Unit, Children’s Hospital, University of Heidelberg, Germany

¹ Correspondence: Universität Heidelberg, Children’s Hospital, Im Neuenheimer Feld 150, 69120 Heidelberg, Germany. E-mail: markus_sperandio{at}med.uni-heidelberg.de

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.

Key Words: leukocyte rolling • capture • shear stress

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