Letter | Published:

‘Slings’ enable neutrophil rolling at high shear

Nature volume 488, pages 399403 (16 August 2012) | Download Citation

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Abstract

Most leukocytes can roll along the walls of venules at low shear stress (1 dyn cm−2), but neutrophils have the ability to roll at tenfold higher shear stress in microvessels in vivo1,2. The mechanisms involved in this shear-resistant rolling are known to involve cell flattening3 and pulling of long membrane tethers at the rear4,5,6. Here we show that these long tethers do not retract as postulated6,7, but instead persist and appear as ‘slings’ at the front of rolling cells. We demonstrate slings in a model of acute inflammation in vivo and on P-selectin in vitro, where P-selectin-glycoprotein-ligand-1 (PSGL-1) is found in discrete sticky patches whereas LFA-1 is expressed over the entire length on slings. As neutrophils roll forward, slings wrap around the rolling cells and undergo a step-wise peeling from the P-selectin substrate enabled by the failure of PSGL-1 patches under hydrodynamic forces. The ‘step-wise peeling of slings’ is distinct from the ‘pulling of tethers’ reported previously4,5,6,8. Each sling effectively lays out a cell-autonomous adhesive substrate in front of neutrophils rolling at high shear stress during inflammation.

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References

  1. 1.

    & Leukocyte margination and deformation in mesenteric venules of rat. Am. J. Physiol. Heart Circ. Physiol. 256, H1667–H1674 (1989)

  2. 2.

    , , , & Biomechanics of leukocyte rolling. Biorheology 48, 1–35 (2011)

  3. 3.

    , , & Variation in the velocity, deformation, and adhesion energy density of leukocytes rolling within venules. Circ. Res. 79, 1122–1130 (1996)

  4. 4.

    et al. Quantitative dynamic footprinting microscopy reveals mechanisms of neutrophil rolling. Nature Methods 7, 821–824 (2010)

  5. 5.

    , , , & Dynamic alterations of membrane tethers stabilize leukocyte rolling on P-selectin. Proc. Natl Acad. Sci. USA 101, 13519–13524 (2004)

  6. 6.

    & Direct observation of membrane tethers formed during neutrophil attachment to platelets or P-selectin under physiological flow. J. Cell Biol. 149, 719–730 (2000)

  7. 7.

    in Leukocyte Adhesion (ed. ) Ch. 2, 25–45 (Academic, 2009).

  8. 8.

    , & Cell protrusions and tethers: a unified approach. Biophys. J. 100, 1697–1707 (2011)

  9. 9.

    , & Cell adhesion molecule distribution relative to neutrophil surface topography assessed by TIRFM. Biophys. J. 97, 379–387 (2009)

  10. 10.

    et al. P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin. J. Cell Biol. 128, 661–671 (1995)

  11. 11.

    in Leukocyte Adhesion (ed. ) Ch. 1, 3–24 (Academic, 2009).

  12. 12.

    et al. Live cell imaging of paxillin in rolling neutrophils by dual-color quantitative dynamic footprinting. Microcirculation 18, 361–372 (2011)

  13. 13.

    , , , & Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages. Blood 96, 719–726 (2000)

  14. 14.

    et al. Immature mouse dendritic cells enter inflamed tissue, a process that requires E- and P-selectin, but not P-selectin glycoprotein ligand 1. Blood 99, 946–956 (2002)

  15. 15.

    Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76, 301–314 (1994)

  16. 16.

    et al. Characterization of murine intercellular adhesion molecule-2. J. Immunol. 156, 4909–4914 (1996)

  17. 17.

    , & Spleen tyrosine kinase Syk is necessary for E-selectin-induced αLβ2 integrin-mediated rolling on intercellular adhesion molecule-1. Immunity 26, 773–783 (2007)

  18. 18.

    & Leukocytes roll on a selectin at physiological flow rates: distinction from and prerequisite for adhesion through integrins. Cell 65, 859–873 (1991)

  19. 19.

    , , , & Rolling on E- or P-selectin induces the extended but not high-affinity conformation of LFA-1 in neutrophils. Blood 116, 617–624 (2010)

  20. 20.

    et al. The junctional adhesion molecule JAM-C regulates polarized transendothelial migration of neutrophils in vivo. Nature Immunol. 12, 761–769 (2011)

  21. 21.

    et al. ICAM-2 mediates neutrophil transmigration in vivo: evidence for stimulus specificity and a role in PECAM-1-independent transmigration. Blood 107, 4721–4727 (2006)

  22. 22.

    et al. Direct observation of catch bonds involving cell-adhesion molecules. Nature 423, 190–193 (2003)

  23. 23.

    et al. P-selectin glycoprotein ligand-1-deficient mice have impaired leukocyte tethering to E-selectin under flow. J. Clin. Invest. 109, 939–950 (2002)

  24. 24.

    et al. Relative contribution of LFA-1 and Mac-1 to neutrophil adhesion and migration. J. Immunol. 163, 5029–5038 (1999)

  25. 25.

    , , , & Stimulated secretion of endothelial von Willebrand factor is accompanied by rapid redistribution to the cell surface of the intracellular granule membrane protein GMP-140. J. Biol. Chem. 264, 7768–7771 (1989)

  26. 26.

    et al. CD63 is an essential cofactor to leukocyte recruitment by endothelial P-selectin. Blood 118, 4265–4273 (2011)

  27. 27.

    , , , & Rolling of Th1 cells via P-selectin glycoprotein ligand-1 stimulates LFA-1-mediated cell binding to ICAM-1. J. Immunol. 174, 1424–1432 (2005)

  28. 28.

    et al. Nitric oxide generated by nNOS in the macula densa regulates the afferent arteriolar diameter in rat kidney. Med. Electron Microsc. 37, 236–241 (2004)

  29. 29.

    et al. Sequential contribution of L- and P-selectin to leukocyte rolling in vivo. J. Exp. Med. 181, 669–675 (1995)

  30. 30.

    et al. Absence of trauma-induced leukocyte rolling in mice deficient in both P-selectin and intercellular adhesion molecule 1. J. Exp. Med. 183, 57–65 (1996)

  31. 31.

    et al. Severe impairment of leukocyte rolling in venules of core 2 glucosaminyltransferase-deficient mice. Blood 97, 3812–3819 (2001)

  32. 32.

    Leukocyte biophysics. An invited review. Cell Biophys. 17, 107–135 (1990)

  33. 33.

    & in Leukocyte Adhesion (ed. ) Ch. 8 221–296 (Academic, 2009)

  34. 34.

    , , & Hydrodynamic narrowing of tubes extruded from cells. Proc. Natl Acad. Sci. USA 103, 7660–7663 (2006)

  35. 35.

    , & Slow viscous motion of a sphere parallel to a plane wall. II. Couette flow. Chem. Eng. Sci. 22, 653–660 (1967)

  36. 36.

    , , & Event-tracking model of adhesion identifies load-bearing bonds in rolling leukocytes. Microcirculation 16, 115–130 (2009)

  37. 37.

    & A semianalytic model of leukocyte rolling. Biophys. J. 87, 2919–2930 (2004)

  38. 38.

    , & Slow viscous motion of a sphere parallel to a plane wall. I. Motion through a quiescent fluid. Chem. Eng. Sci. 22, 637–651 (1967)

  39. 39.

    New Statistical Procedures For The Social Sciences: Modern Solutions To Basic Problems (Lawrence Erlbaum Associates, 1987)

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Acknowledgements

The authors thank A. Zychlinsky for comments and reading the manuscript. This study was supported by the NCRP-Scientist Development Grant 11SDG7340005 from the American Heart Association (P.S.), WSA postdoctoral fellowship 10POST4160142-01 from American Heart Association (E.K.K.) and NIH EB 02185 (K.L.).

Author information

Affiliations

  1. Division of Inflammation Biology, La Jolla Institute for Allergy and Immunology, La Jolla, California 92037, USA

    • Prithu Sundd
    • , Ekaterina K. Koltsova
    • , Yoshihiro Kuwano
    • , Maria K. Pospieszalska
    •  & Klaus Ley
  2. Department of Physics, University of California San Diego, La Jolla, California 92093, USA

    • Edgar Gutierrez
    •  & Alex Groisman
  3. Department of Dermatology, University of Tokyo, Tokyo 113-8655, Japan

    • Yoshihiro Kuwano
  4. Laboratory of Electron Microscopy, University of Tokyo, Tokyo 113-8655, Japan

    • Satoru Fukuda

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Contributions

P.S. performed all the experiments and image analysis. E.G. and A.G. designed the microfluidic device. M.K.P. calculated the fraction of bond force and torque shared by slings and tethers. E.K.K. was involved in culturing of Th1 CD4 T cells. Y.K. and S.F performed the scanning electron microscopy. P.S. and K.L. wrote the manuscript. K.L. supervised the project. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Klaus Ley.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-28, Supplementary Notes 1-4 and legends for Supplementary Movies 1-9.

Videos

  1. 1.

    Supplementary Movie 1

    Slings formed by DiI-stained mouse bone marrow neutrophil rolling on P-selectin - see Supplementary Information file for full legend.

  2. 2.

    Supplementary Movie 2

    Sling formed by an EGFP neutrophil rolling on P-selectin in whole blood of Lyz2-EGFP mouse - see Supplementary Information file for full legend.

  3. 3.

    Supplementary Movie 3

    Wrapping of slings around a DiI-stained mouse bone marrow neutrophil rolling on P-selectin - see Supplementary Information file for full legend.

  4. 4.

    Supplementary Movie 4

    Wrapping of sling by a leukocyte rolling in the cremaster venule of a WT mouse - see Supplementary Information file for full legend.

  5. 5.

    Supplementary Movie 5

    Sling formation by a leukocyte rolling in the cremaster venule of a WT mouse. Image processed to reveal sling - see Supplementary Information file for full legend.

  6. 6.

    Supplementary Movie 6

    Tether (arrowhead) swings over to become a sling (arrow) - see Supplementary Information file for full legend.

  7. 7.

    Supplementary Movie 7

    Tether (arrowhead) swings over to become a sling (arrow) - see Supplementary Information file for full legend.

  8. 8.

    Supplementary Movie 8

    Step-wise peeling of a sling. PSGL-1 patches (red spots) visible on sling (green) - see Supplementary Information file for full legend.

  9. 9.

    Supplementary Movie 9

    Step-wise peeling of a sling. PSGL-1 patches (red spots) visible on sling (green) - see Supplementary Information file for full legend.

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DOI

https://doi.org/10.1038/nature11248

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