P-selectin glycoprotein ligand-1–deficient mice have impaired leukocyte tethering to E-selectin under flow
Lijun Xia, … , Klaus Ley, Rodger P. McEver
Lijun Xia, … , Klaus Ley, Rodger P. McEver
Published April 1, 2002
Citation Information: J Clin Invest. 2002;109(7):939-950. https://doi.org/10.1172/JCI14151.
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Article Immunology

P-selectin glycoprotein ligand-1–deficient mice have impaired leukocyte tethering to E-selectin under flow

Abstract

P-selectin glycoprotein ligand-1 (PSGL-1) mediates rolling of leukocytes on P-selectin under flow. The glycoproteins that enable leukocyte tethering to or rolling on E-selectin are not known. We used gene targeting to prepare PSGL-1–deficient (PSGL-1–/–) mice, which were healthy but had moderately elevated total blood leukocytes. Fluid-phase E-selectin bound to approximately 70% fewer sites on PSGL-1–/– than PSGL-1+/+ neutrophils. Compared with PSGL-1+/+ leukocytes, significantly fewer PSGL-1–/– leukocytes rolled on E-selectin in vitro, because their initial tethering to E-selectin was impaired. The residual cells that tethered rolled with the same shear resistance and velocities as PSGL-1+/+ leukocytes. Compared with PSGL-1+/+ mice, significantly fewer PSGL-1–/– leukocytes rolled on E-selectin in TNF-α–treated venules of cremaster muscle in which P-selectin function was blocked by an mAb. The residual PSGL-1–/– leukocytes that tethered rolled with slow velocities equivalent to those of PSGL-1+/+ leukocytes. These results reveal a novel function for PSGL-1 in tethering leukocytes to E-selectin under flow.

Authors

Lijun Xia, Markus Sperandio, Tadayuki Yago, J. Michael McDaniel, Richard D. Cummings, Sonia Pearson-White, Klaus Ley, Rodger P. McEver

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Tethering and rolling of PSGL-1+/+ and PSGL-1–/– leukocytes on E-selecti...
Tethering and rolling of PSGL-1+/+ and PSGL-1–/– leukocytes on E-selectin in vitro. (a) Leukocytes were perfused over mAb-captured E-selectin–IgM or capture mAb only, in the presence or absence of EDTA or anti–E-selectin mAb 9A9. After 5 minutes, the number of rolling cells was quantified. *P < 0.05 vs. PSGL-1+/+. (b) Leukocytes in the presence or absence of anti–L-selectin mAb Mel-14 were perfused over E-selectin–IgM at the indicated wall shear stress. After 5 minutes, the number of rolling cells was quantified. *P < 0.05 vs. PSGL-1+/+ + anti-L. (c) Leukocytes were perfused over E-selectin–IgM at the indicated wall shear stress. The number of cells that tethered to E-selectin during the first 30 seconds was quantified and normalized by dividing by the number of cells delivered across the field of view in the focal plane of the monolayer. Only primary tethers of flowing leukocytes to E-selectin were measured. *P < 0.05 vs. PSGL-1+/+. (d and e) Leukocytes were allowed to accumulate on E-selectin under a wall shear stress of 0.5 dyn/cm2. Wall shear stress was increased every 30 seconds, and the percentage of cells remaining adherent and the rolling velocities of the cells were determined. Data represent mean ± SEM of four experiments.