A novel method to analyze leukocyte rolling behavior in vivo

Leukocyte endothelial cell interaction is a fundamentally important process in many disease states. Current methods to analyze such interactions include the parallel-plate flow chamber and intravital microscopy. Here, we present an improvement of the traditional intravital microscopy that allows leukocyte-endothelial cell interaction to be studied from the time the leukocyte makes its initial contact with the endothelium until it adheres to or detaches from the endothelium. The leukocyte is tracked throughout the venular tree with the aid of a motorized stage and the rolling and adhesive behavior is measured off-line. Because this method can involve human error, methods to automate the tracking procedure have been developed. This novel tracking method allows for a more detailed examination of leukocyte-endothelial cell interactions.

[1]  S. Baez,et al.  An open cremaster muscle preparation for the study of blood vessels by in vivo microscopy. , 1973, Microvascular research.

[2]  J. Forrester,et al.  Adhesion of neutrophil leucocytes under conditions of flow. , 1984, Journal of cell science.

[3]  E. Butcher.  Leukocyte-endothelial cell recognition: Three (or more) steps to specificity and diversity , 1991, Cell.

[4]  A. Beaudet,et al.  Sequential contribution of L- and P-selectin to leukocyte rolling in vivo , 1995, The Journal of experimental medicine.

[5]  E. Kunkel,et al.  Distinct phenotype of E-selectin-deficient mice. E-selectin is required for slow leukocyte rolling in vivo. , 1996, Circulation research.

[6]  G. Nash,et al.  Endothelial-borne platelet-activating factor and interleukin-8 rapidly immobilize rolling neutrophils. , 1997, The American journal of physiology.

[7]  A. Beaudet,et al.  LFA-1 is sufficient in mediating neutrophil emigration in Mac-1-deficient mice. , 1997, The Journal of clinical investigation.

[8]  R. Jain,et al.  Mice Lacking E‐Selection Show Normal Numbers of Rolling Leukocytes but Reduced Leukocyte Stable Arrest on Cytokine‐Activated Microvascular Endothelium , 1998, Microcirculation.

[9]  K E Norman,et al.  Transit time of leukocytes rolling through venules controls cytokine-induced inflammatory cell recruitment in vivo. , 1998, The Journal of clinical investigation.

[10]  E. Butcher,et al.  Chemokines and the arrest of lymphocytes rolling under flow conditions. , 1998, Science.

[11]  K. Ley,et al.  Importance of E-selectin for firm leukocyte adhesion in vivo. , 1998, Circulation research.

[12]  Z. Ding,et al.  Relative contribution of LFA-1 and Mac-1 to neutrophil adhesion and migration. , 1999, Journal of immunology.

[13]  K. Ley,et al.  Mice lacking two or all three selectins demonstrate overlapping and distinct functions for each selectin. , 1999, Journal of immunology.

[14]  E. Butcher,et al.  L-selectin–mediated Leukocyte Adhesion In Vivo: Microvillous Distribution Determines Tethering Efficiency, But Not Rolling Velocity , 1999, The Journal of experimental medicine.

[15]  F. Luscinskas,et al.  MCP-1 and IL-8 trigger firm adhesion of monocytes to vascular endothelium under flow conditions , 1999, Nature.

[16]  E. Kunkel,et al.  Leukocyte Arrest During Cytokine-Dependent Inflammation In Vivo1 , 2000, The Journal of Immunology.

[17]  A. Beaudet,et al.  Severe inflammatory defect and reduced viability in CD18 and E-selectin double-mutant mice. , 2000, The Journal of clinical investigation.

[18]  S. Simon,et al.  Neutrophil Tethering on E-Selectin Activates β2 Integrin Binding to ICAM-1 Through a Mitogen-Activated Protein Kinase Signal Transduction Pathway1 , 2000, The Journal of Immunology.

[19]  K E Norman,et al.  An Effective and Economical Solution for Digitizing and Analyzing Video Recordings of the Microcirculation , 2001, Microcirculation.

[20]  Scott T. Acton,et al.  Tracking leukocytes in vivo with shape and size constrained active contours , 2002, IEEE Transactions on Medical Imaging.

[21]  Jessica L. Dunne,et al.  Control of leukocyte rolling velocity in TNF-alpha-induced inflammation by LFA-1 and Mac-1. , 2002, Blood.

[22]  Klaus Ley,et al.  Automatic tracking of rolling leukocytes in vivo. , 2002, Microvascular research.

[23]  Jessica L. Dunne,et al.  Mac-1, but Not LFA-1, Uses Intercellular Adhesion Molecule-1 to Mediate Slow Leukocyte Rolling in TNF-α-Induced Inflammation 1 , 2003, The Journal of Immunology.

[24]  R. Hotchkiss,et al.  Intravital microscopy comparing T lymphocyte trafficking to the spleen and the mesenteric lymph node. , 2003, American journal of physiology. Heart and circulatory physiology.

[25]  D. Broide,et al.  A murine model to study leukocyte rolling and intravascular trafficking in lung microvessels. , 2003, The American journal of pathology.

[26]  Janet Liversidge,et al.  Reduction in shear stress, activation of the endothelium, and leukocyte priming are all required for leukocyte passage across the blood—retina barrier , 2004, Journal of leukocyte biology.

[27]  Scott T. Acton,et al.  Vessel boundary tracking for intravital microscopy via multiscale gradient vector flow snakes , 2004, IEEE Transactions on Biomedical Engineering.

[28]  Scott T. Acton,et al.  Image stabilization and registration for tracking cells in the microvasculature , 2005, IEEE Transactions on Biomedical Engineering.