Analysis of Integrin-Dependent Rapid Adhesion Under Laminar-Flow Conditions

  • Carlo Laudanna
Part of the Methods in Molecular Biology™ book series (MIMB, volume 239)


Adhesion molecules mediate recognition of the blood vessels by circulating leukocytes and support their selective targeting to different organs (1). In the vessels, the blood flow imposes peculiar conditions by generating a wall shear stress that opposes leukocyte stable arrest on the endothelium. As the rapidness of integrin activation is mandatory to leukocyte adhesion to the blood vessels, any analysis of adhesion triggering relevant to leukocyte in vivo migration should be performed under flow conditions. Here, a method is illustrated to quantitatively analyze the rapid induction of integrin-dependent lymphocyte adhesion by chemokines under flow conditions. Glass capillary tubes are cocoated with purified ligands for selectins and integrins and with chemokines, thus reconstituting the minimal requirement to support tethering, rolling, and arrest under flow conditions, with a physiologic wall shear stress of 2 dynes/cm2. PNAd, ICAM-1, and the chemokine CCL21 are used as paradigmatic adhesion molecules and physiologic proadhesive agonist. Naive lymphocytes isolated from mouse lymph nodes are used as the cell model. The procedure for quantitative analysis is discussed.


Wall Shear Stress Mouse Lymph Node Glass Capillary Tube Integrin Activation Human Tonsil 
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  1. 1.
    Butcher, E. C., Williams, M., Youngman, K., Rott, L., and Briskin, M. (1999) Lymphocyte trafficking and regional immunity. Adv. Immunol. 72, 209–253.PubMedCrossRefGoogle Scholar
  2. 2.
    Streeter, P. R., Rouse, B. T. N., and Butcher, E. C. (1999) Immunohistologic and functional characterization of a vascular addressin involved in lymphocyte homing into peripheral lymph nodes. J. Cell. Biol. 107, 1853–1862.CrossRefGoogle Scholar
  3. 3.
    Campbell, J. J., Hedrick, J., Zlotnik, A., Siani, M. A., Thompson, D. A., and Butcher, E. C. (1998) Chemokines and the arrest of lymphocytes rolling under flow conditions. Science 279, 381–384.PubMedCrossRefGoogle Scholar
  4. 4.
    Constantin, G., Majeed, M., Giagulli, C., et al. (2000) Chemokines trigger immediate β2 integrin affinity and mobility changes: differential regulation and roles in lymphocyte arrest under flow. Immunity 13, 759–769.PubMedCrossRefGoogle Scholar
  5. 5.
    Norman, K. E., Katapodis, A. G., Thoma, G., et al. (2000) P-Selectin glycoprotein ligand-1 supports rolling on E-and P-selectin in vivo. Blood 96, 3585–3593.PubMedGoogle Scholar
  6. 6.
    Lawrence, M. B. and Springer, T. A. (1991) Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion through integrins. Cell 65, 859–873.PubMedCrossRefGoogle Scholar
  7. 7.
    Grabovsky, V., Feigelson, S., Chen, C., et al. (2000) Subsecond induction of α4 integrin clustering by immobilized chemokines stimulates leukocyte tethering and rolling on endothelial vascular cell adhesion molecule 1 under flow conditions. J. Exp. Med. 192, 495–506.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Carlo Laudanna
    • 1
  1. 1.Department of Pathology, Faculty of MedicineUniversity of VeronaVeronaItaly

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