Thromb Haemost 2007; 97(05): 814-821
DOI: 10.1160/TH06-08-0439
Platelets and Blood Cells
Schattauer GmbH

Sequential adhesion of platelets and leukocytes from flowing whole blood onto a collagen-coated surface: Requirement for a GpVI-binding site in collagen

Lynn M. Butler
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
Tom Metson-Scott
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
Jo Felix
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
Anita Abhyankar
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
G. Ed Rainger
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
Richard W. Farndale
2   Department of Biochemistry, University of Cambridge, Cambridge, UK
,
Stephen P. Watson
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
,
Gerard B. Nash
1   Centre for Cardiovascular Science, The Medical School, University of Birmingham, Birmingham, UK
› Author Affiliations
Financial support: GER was supported by a British Heart Foundation Non-Clinical Senior Lectureship.
Further Information

Publication History

Received 10 August 2006

Accepted after resubmission 08 February 2007

Publication Date:
24 November 2017 (online)

Summary

The adhesion of leukocytes to immobilised platelets may contribute to inflammatory and thrombotic responses in damaged tissue. To investigate the conditions under which platelets and leukocytes might be deposited together in vessels, we perfused fluorescently-labelled whole blood through glass capillaries coated with various collagen preparations. Video-microscopic observations of the surface showed that platelets formed numerous, individual, rolling and stationary attachments to surfaces coated with acid-soluble, monomeric collagen. However, leukocyte interactions with the deposited platelets were rare. If the blood was washed out, the adherent platelets became more activated, and many rolling adherent leukocytes were observed if a second bolus of blood was perfused over them. This suggested that platelet activation had initially been inadequate to support leukocyte capture. Next, fibrillar collagen was adsorbed to the capillaries to present an ordered array of peptide motifs to platelet receptor glycoprotein (Gp)VI and transduce an activating signal. In this case, platelets were deposited in discrete, stable aggregates and the bound platelets captured many flowing leukocytes.Alternatively, acid-soluble collagen was seeded with collagen-related peptide (CRP) known to contain a GpVI-binding motif.Again, platelet adhesion became stable, and numerous flowing leukocytes were captured.Addition of antibody against GpVI or against P-selectin greatly reduced leukocyte adhesion to the platelets. Thus, in whole blood, platelets binding to exposed collagen need to be activated through GpVI in order to expose sufficient P-selectin to allow efficient capture of flowing leukocytes to take place.

 
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