Platelet/Polymorphonuclear Leukocyte Interaction: P-Selectin Triggers Protein-Tyrosine Phosphorylation–Dependent CD11b/CD18 Adhesion: Role of PSGL-1 as a Signaling Molecule

Polymorphonuclear leukocyte (PMN) adhesion to activated platelets is important for the recruitment of PMN at sites of vascular damage and thrombus formation. We have recently shown that binding of activated platelets to PMN in mixed cell suspensions under shear involves P-selectin and the activated β2-integrin CD11b/CD18. Integrin activation required signaling mechanisms that were sensitive to tyrosine kinase inhibitors.1 Here we show that mixing activated, paraformaldehyde (PFA)-fixed platelets with PMNs under shear conditions leads to rapid and fully reversible tyrosine phosphorylation of a prominent protein of 110 kD (P∼110). Phosphorylation was both Ca2+ and Mg2+ dependent and was blocked by antibodies against P-selectin or CD11b/CD18, suggesting that both adhesion molecules need to engage with their respective ligands to trigger phosphorylation of P∼110. The inhibition of P∼110 phosphorylation by tyrosine kinase inhibitors correlates with the inhibition of platelet/PMN aggregation. Similar effects were observed when platelets were substituted by P-selectin–transfected Chinese hamster ovary (CHO-P) cells or when PMN were stimulated with P-selectin–IgG fusion protein. CHO-P/PMN mixed-cell aggregation and P-selectin–IgG–triggered PMN/PMN aggregation as well as P∼110 phosphorylation were all blocked by antibodies against P-selectin or CD18. In each case PMN adhesion was sensitive to the tyrosine kinase inhibitor genistein. The antibody PL-1 against P-selectin glycoprotein ligand-1 (PSGL-1) blocked platelet/PMN aggregation, indicating that PSGL-1 was the major tethering ligand for P-selectin in this experimental system. Moreover, engagement of PSGL-1 with a nonadhesion blocking antibody triggered β2-integrin–dependent genistein-sensitive aggregation as well as tyrosine phosphorylation in PMN. This study shows that binding of P-selectin to PSGL-1 triggers tyrosine kinase–dependent mechanisms that lead to CD11b/CD18 activation in PMN. The availability of the β2-integrin to engage with its ligands on the neighboring cells is necessary for the tyrosine phosphorylation of P∼110.

[1]  A. King,et al.  A juxtacrine mechanism for neutrophil adhesion on platelets involves platelet-activating factor and a selectin-dependent activation process. , 1998, Blood.

[2]  D. Vestweber,et al.  Stimulation of P‐selectin glycoprotein ligand‐1 on mouse neutrophils activates β2 ‐integrin mediated cell attachment to ICAM‐1 , 1998, European journal of immunology.

[3]  C. Haslett,et al.  Potential pro‐inflammatory effects of soluble E‐selectin upon neutrophil function , 1998, European journal of immunology.

[4]  A. Weyrich,et al.  Engagement of P-selectin Glycoprotein Ligand-1 Enhances Tyrosine Phosphorylation and Activates Mitogen-activated Protein Kinases in Human Neutrophils* , 1997, The Journal of Biological Chemistry.

[5]  T. Springer,et al.  Neutrophil accumulation on activated, surface-adherent platelets in flow is mediated by interaction of Mac-1 with fibrinogen bound to alphaIIbbeta3 and stimulated by platelet-activating factor. , 1997, The Journal of clinical investigation.

[6]  C. Gahmberg Leukocyte adhesion: CD11/CD18 integrins and intercellular adhesion molecules. , 1997, Current opinion in cell biology.

[7]  S. Sharar,et al.  Binding of human peripheral blood polymorphonuclear leukocytes to E-selectin (CD62E) does not promote their activation. , 1997, Journal of immunology.

[8]  D. Kirchhofer,et al.  Specific accumulation of circulating monocytes and polymorphonuclear leukocytes on platelet thrombi in a vascular injury model. , 1997, Blood.

[9]  C. Cerletti,et al.  Platelet/polymorphonuclear leukocyte interaction in dynamic conditions: evidence of adhesion cascade and cross talk between P-selectin and the beta 2 integrin CD11b/CD18. , 1996, Blood.

[10]  C. Cerletti,et al.  Thrombin-activated Human Platelets Release two NAP-2 Variants that Stimulate Polymorphonuclear Leukocytes , 1996, Thrombosis and Haemostasis.

[11]  G. Zimmerman,et al.  Adhesion and signaling in vascular cell-cell interactions. , 1996, The Journal of clinical investigation.

[12]  G. Zimmerman,et al.  Perspectives series: Cell adhesion in vascular biology. Adhesion and signaling in vascular cell-cell interactions , 1996 .

[13]  T. Springer,et al.  Neutrophil rolling, arrest, and transmigration across activated, surface-adherent platelets via sequential action of P-selectin and the beta 2-integrin CD11b/CD18. , 1996, Blood.

[14]  G. Nash,et al.  Continuous activation and deactivation of integrin CD11b/CD18 during de novo expression enables rolling neutrophils to immobilize on platelets. , 1996, Blood.

[15]  K. Ley,et al.  β2 integrins mediate protein tyrosine phosphorylation in human neutrophils , 1996, Journal of leukocyte biology.

[16]  L. Koenderman,et al.  Platelet-dependent primary hemostasis promotes selectin- and integrin-mediated neutrophil adhesion to damaged endothelium under flow conditions. , 1996, Blood.

[17]  A. Weyrich,et al.  Activated platelets signal chemokine synthesis by human monocytes. , 1996, The Journal of clinical investigation.

[18]  C. Gahmberg,et al.  Intercellular adhesion molecule-2 (CD102) binds to the leukocyte integrin CD11b/CD18 through the A domain. , 1995, Journal of immunology.

[19]  G. Zimmerman,et al.  P-selectin regulates platelet-activating factor synthesis and phagocytosis by monocytes. , 1995, Journal of immunology.

[20]  B. Furie,et al.  The Molecular Basis of Platelet and Endothelial Cell Interaction with Neutrophils and Monocytes: Role of P-Selectin and the P-Selectin Ligand, PSGL-1 , 1995, Thrombosis and Haemostasis.

[21]  G. Zimmerman,et al.  Activation of polymorphonuclear leukocytes reduces their adhesion to P-selectin and causes redistribution of ligands for P-selectin on their surfaces. , 1995, The Journal of clinical investigation.

[22]  A. Weyrich,et al.  Monocyte tethering by P-selectin regulates monocyte chemotactic protein-1 and tumor necrosis factor-alpha secretion. Signal integration and NF-kappa B translocation. , 1995, The Journal of clinical investigation.

[23]  R. Cummings,et al.  P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin , 1995, The Journal of cell biology.

[24]  C. Power,et al.  Cloning of a full-length cDNA encoding the neutrophil-activating peptide ENA-78 from human platelets. , 1994, Gene.

[25]  Joseph F. Murphy,et al.  The vitronectin receptor (alpha v beta 3) is implicated, in cooperation with P-selectin and platelet-activating factor, in the adhesion of monocytes to activated endothelial cells. , 1994, The Biochemical journal.

[26]  S. Watson,et al.  Cell surface P- and E-selectin support shear-dependent rolling of bovine gamma/delta T cells. , 1994, Journal of immunology.

[27]  T. Carlos,et al.  Leukocyte-endothelial adhesion molecules. , 1994, Blood.

[28]  M. Vadas,et al.  P-selectin interacts with a beta 2-integrin to enhance phagocytosis. , 1994, Journal of immunology.

[29]  G. Pellegrini,et al.  P-selectin induces the expression of tissue factor on monocytes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[30]  T. Springer,et al.  A functional integrin ligand on the surface of platelets: intercellular adhesion molecule-2. , 1994, The Journal of clinical investigation.

[31]  C. Laudanna,et al.  Beta 2 integrin-dependent protein tyrosine phosphorylation and activation of the FGR protein tyrosine kinase in human neutrophils , 1994, The Journal of cell biology.

[32]  Timothy A. Springer,et al.  The dynamic regulation of integrin adhesiveness , 1994, Current Biology.

[33]  B. Furie,et al.  Expression cloning of a functional glycoprotein ligand for P-selectin , 1993, Cell.

[34]  T. Irimura,et al.  Activated platelets induce superoxide anion release by monocytes and neutrophils through P-selectin (CD62). , 1993, Journal of immunology.

[35]  M. Hahne,et al.  Five tumor necrosis factor-inducible cell adhesion mechanisms on the surface of mouse endothelioma cells mediate the binding of leukocytes , 1993, The Journal of cell biology.

[36]  D. Altieri,et al.  The structural motif glycine 190-valine 202 of the fibrinogen gamma chain interacts with CD11b/CD18 integrin (alpha M beta 2, Mac-1) and promotes leukocyte adhesion. , 1993, The Journal of biological chemistry.

[37]  C. Benjamin,et al.  Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets , 1992, Nature.

[38]  D. Wagner,et al.  P-selectin and E-selectin. Distinct but overlapping leukocyte ligand specificities. , 1992, The Journal of biological chemistry.

[39]  G. Zimmerman,et al.  Coexpression of GMP-140 and PAF by endothelium stimulated by histamine or thrombin: a juxtacrine system for adhesion and activation of neutrophils , 1991, The Journal of cell biology.

[40]  T. Kuijpers,et al.  Role of endothelial leukocyte adhesion molecule-1 and platelet-activating factor in neutrophil adherence to IL-1-prestimulated endothelial cells. Endothelial leukocyte adhesion molecule-1-mediated CD18 activation. , 1991, Journal of immunology.

[41]  S. Wright,et al.  Endothelial-leukocyte adhesion molecule 1 stimulates the adhesive activity of leukocyte integrin CR3 (CD11b/CD18, Mac-1, alpha m beta 2) on human neutrophils , 1991, The Journal of experimental medicine.

[42]  J. Gamble,et al.  Adhesion protein GMP140 inhibits superoxide anion release by human neutrophils. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[43]  D. Altieri,et al.  A unique recognition site mediates the interaction of fibrinogen with the leukocyte integrin Mac-1 (CD11b/CD18). , 1990, The Journal of biological chemistry.

[44]  E. Butcher,et al.  Identification of a human peripheral lymph node homing receptor: a rapidly down-regulated adhesion molecule. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. Baggiolini,et al.  A novel cleavage product of beta-thromboglobulin formed in cultures of stimulated mononuclear cells activates human neutrophils. , 1989, Biochemical and biophysical research communications.

[46]  C. Cabañas,et al.  Characterization of a CD11c-reactive monoclonal antibody (HC1/1) obtained by immunizing with phorbol ester differentiated U937 cells. , 1988, Hybridoma.

[47]  T. Springer,et al.  Contributions of the Mac-1 glycoprotein family to adherence-dependent granulocyte functions: structure-function assessments employing subunit-specific monoclonal antibodies. , 1986, Journal of immunology.

[48]  E. Dejana,et al.  Platelet Adhesion to Subendothelium - Effect of Shear Rate, Hematocrit and Platelet Count on the Dynamic Equilibrium Between Platelets Adhering to and Detaching from the Surface , 1985, Thrombosis and Haemostasis.

[49]  H. Ochs,et al.  The role of neutrophil membrane glycoprotein GP-150 in neutrophil adherence to endothelium in vitro. , 1985, Blood.

[50]  J. Strominger,et al.  Three distinct antigens associated with human T-lymphocyte-mediated cytolysis: LFA-1, LFA-2, and LFA-3. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[51]  G. Bazzoni,et al.  Are changes in integrin affinity and conformation overemphasized? , 1998, Trends in biochemical sciences.

[52]  K Konstantopoulos,et al.  Perspectives Series: Cell Adhesion in Vascular Biology Effects of Fluid Dynamic Forces on Vascular Cell Adhesion , 1996 .

[53]  I. Feuerstein,et al.  Role of P-selectin and leukocyte activation in polymorphonuclear cell adhesion to surface adherent activated platelets under physiologic shear conditions (an injury vessel wall model). , 1994, Blood.

[54]  S. Simon,et al.  Beta 2-integrin and L-selectin are obligatory receptors in neutrophil aggregation. , 1993, Blood.