Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin

Konstantinos Konstantopoulos, Sriram Neelamegham, Alan R. Burns, Eric Hentzen, Geoffrey S. Kansas, Karen R. Snapp, Ellen L. Berg, J. David Hellums, C. Wayne Smith, Larry V. McIntire, Scott I. Simon

Research output: Contribution to journalArticle

133 Citations (Scopus)

Abstract

Background - After activation, platelets adhere to neutrophils via P- selectin and β2-integrin. The molecular mechanisms and adhesion events in whole blood exposed to venous levels of hydrodynamic shear in the absence of exogenous activation remain unknown. Methods and Results - Whole blood was sheared at ≃100 s-1. The kinetics of neutrophil-platelet adhesion and neutrophil aggregation were measured in real time by flow cytometry. P- selectin was upregulated to the platelet surface in response to shear and was the primary factor mediating neutrophil-platelet adhesion. The extent of neutrophil aggregation increased linearly with platelet adhesion to neutrophils. Blocking either P-selectin, its glycoprotein ligand PSGL-1, or both simultaneously by preincubation with a monoclonal antibody resulted in equivalent inhibition of neutrophil-platelet adhesion (≃30%) and neutrophil aggregation (≃70%). The residual amount of neutrophil adhesion was blocked with anti-CD11b/CD18. Treatment of blood with prostacyclin analogue ZK36374, which raises cAMP levels in platelets, blocked P-selectin upregulation and neutrophil aggregation to baseline. Complete abrogation of platelet- neutrophil adhesion required both ZK36374 and anti-CD18. Electron microscopic observations of fixed blood specimens revealed that platelets augmented neutrophil aggregation both by forming bridges between neutrophils and through contact-mediated activation. Conclusions - The results are consistent with a model in which venous levels of shear support platelet adherence to neutrophils via P-selectin binding PSGL-1. This interaction alone is sufficient to mediate neutrophil aggregation. Abrogation of platelet adhesion and aggregation requires blocking Mac-1 in addition to PSGL-1 or P-selectin. The described mechanisms are likely of key importance in the pathogenesis and progression of thrombotic disorders that are exacerbated by leukocyte- platelet aggregation.

Original languageEnglish (US)
Pages (from-to)873-882
Number of pages10
JournalCirculation
Volume98
Issue number9
StatePublished - Sep 1 1998
Externally publishedYes

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P-Selectin
Integrins
Neutrophils
Blood Platelets
Platelet Aggregation
Leukocyte Disorders
Platelet Activation
Epoprostenol
Hydrodynamics

Keywords

  • Blood cells
  • Glycoproteins
  • Integrins
  • Neutrophils
  • Platelets

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Konstantopoulos, K., Neelamegham, S., Burns, A. R., Hentzen, E., Kansas, G. S., Snapp, K. R., ... Simon, S. I. (1998). Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin. Circulation, 98(9), 873-882.

Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin. / Konstantopoulos, Konstantinos; Neelamegham, Sriram; Burns, Alan R.; Hentzen, Eric; Kansas, Geoffrey S.; Snapp, Karen R.; Berg, Ellen L.; Hellums, J. David; Smith, C. Wayne; McIntire, Larry V.; Simon, Scott I.

In: Circulation, Vol. 98, No. 9, 01.09.1998, p. 873-882.

Research output: Contribution to journalArticle

Konstantopoulos, K, Neelamegham, S, Burns, AR, Hentzen, E, Kansas, GS, Snapp, KR, Berg, EL, Hellums, JD, Smith, CW, McIntire, LV & Simon, SI 1998, 'Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin', Circulation, vol. 98, no. 9, pp. 873-882.
Konstantopoulos K, Neelamegham S, Burns AR, Hentzen E, Kansas GS, Snapp KR et al. Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin. Circulation. 1998 Sep 1;98(9):873-882.
Konstantopoulos, Konstantinos ; Neelamegham, Sriram ; Burns, Alan R. ; Hentzen, Eric ; Kansas, Geoffrey S. ; Snapp, Karen R. ; Berg, Ellen L. ; Hellums, J. David ; Smith, C. Wayne ; McIntire, Larry V. ; Simon, Scott I. / Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and β2-integrin. In: Circulation. 1998 ; Vol. 98, No. 9. pp. 873-882.
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abstract = "Background - After activation, platelets adhere to neutrophils via P- selectin and β2-integrin. The molecular mechanisms and adhesion events in whole blood exposed to venous levels of hydrodynamic shear in the absence of exogenous activation remain unknown. Methods and Results - Whole blood was sheared at ≃100 s-1. The kinetics of neutrophil-platelet adhesion and neutrophil aggregation were measured in real time by flow cytometry. P- selectin was upregulated to the platelet surface in response to shear and was the primary factor mediating neutrophil-platelet adhesion. The extent of neutrophil aggregation increased linearly with platelet adhesion to neutrophils. Blocking either P-selectin, its glycoprotein ligand PSGL-1, or both simultaneously by preincubation with a monoclonal antibody resulted in equivalent inhibition of neutrophil-platelet adhesion (≃30{\%}) and neutrophil aggregation (≃70{\%}). The residual amount of neutrophil adhesion was blocked with anti-CD11b/CD18. Treatment of blood with prostacyclin analogue ZK36374, which raises cAMP levels in platelets, blocked P-selectin upregulation and neutrophil aggregation to baseline. Complete abrogation of platelet- neutrophil adhesion required both ZK36374 and anti-CD18. Electron microscopic observations of fixed blood specimens revealed that platelets augmented neutrophil aggregation both by forming bridges between neutrophils and through contact-mediated activation. Conclusions - The results are consistent with a model in which venous levels of shear support platelet adherence to neutrophils via P-selectin binding PSGL-1. This interaction alone is sufficient to mediate neutrophil aggregation. Abrogation of platelet adhesion and aggregation requires blocking Mac-1 in addition to PSGL-1 or P-selectin. The described mechanisms are likely of key importance in the pathogenesis and progression of thrombotic disorders that are exacerbated by leukocyte- platelet aggregation.",
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AU - Konstantopoulos, Konstantinos

AU - Neelamegham, Sriram

AU - Burns, Alan R.

AU - Hentzen, Eric

AU - Kansas, Geoffrey S.

AU - Snapp, Karen R.

AU - Berg, Ellen L.

AU - Hellums, J. David

AU - Smith, C. Wayne

AU - McIntire, Larry V.

AU - Simon, Scott I.

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N2 - Background - After activation, platelets adhere to neutrophils via P- selectin and β2-integrin. The molecular mechanisms and adhesion events in whole blood exposed to venous levels of hydrodynamic shear in the absence of exogenous activation remain unknown. Methods and Results - Whole blood was sheared at ≃100 s-1. The kinetics of neutrophil-platelet adhesion and neutrophil aggregation were measured in real time by flow cytometry. P- selectin was upregulated to the platelet surface in response to shear and was the primary factor mediating neutrophil-platelet adhesion. The extent of neutrophil aggregation increased linearly with platelet adhesion to neutrophils. Blocking either P-selectin, its glycoprotein ligand PSGL-1, or both simultaneously by preincubation with a monoclonal antibody resulted in equivalent inhibition of neutrophil-platelet adhesion (≃30%) and neutrophil aggregation (≃70%). The residual amount of neutrophil adhesion was blocked with anti-CD11b/CD18. Treatment of blood with prostacyclin analogue ZK36374, which raises cAMP levels in platelets, blocked P-selectin upregulation and neutrophil aggregation to baseline. Complete abrogation of platelet- neutrophil adhesion required both ZK36374 and anti-CD18. Electron microscopic observations of fixed blood specimens revealed that platelets augmented neutrophil aggregation both by forming bridges between neutrophils and through contact-mediated activation. Conclusions - The results are consistent with a model in which venous levels of shear support platelet adherence to neutrophils via P-selectin binding PSGL-1. This interaction alone is sufficient to mediate neutrophil aggregation. Abrogation of platelet adhesion and aggregation requires blocking Mac-1 in addition to PSGL-1 or P-selectin. The described mechanisms are likely of key importance in the pathogenesis and progression of thrombotic disorders that are exacerbated by leukocyte- platelet aggregation.

AB - Background - After activation, platelets adhere to neutrophils via P- selectin and β2-integrin. The molecular mechanisms and adhesion events in whole blood exposed to venous levels of hydrodynamic shear in the absence of exogenous activation remain unknown. Methods and Results - Whole blood was sheared at ≃100 s-1. The kinetics of neutrophil-platelet adhesion and neutrophil aggregation were measured in real time by flow cytometry. P- selectin was upregulated to the platelet surface in response to shear and was the primary factor mediating neutrophil-platelet adhesion. The extent of neutrophil aggregation increased linearly with platelet adhesion to neutrophils. Blocking either P-selectin, its glycoprotein ligand PSGL-1, or both simultaneously by preincubation with a monoclonal antibody resulted in equivalent inhibition of neutrophil-platelet adhesion (≃30%) and neutrophil aggregation (≃70%). The residual amount of neutrophil adhesion was blocked with anti-CD11b/CD18. Treatment of blood with prostacyclin analogue ZK36374, which raises cAMP levels in platelets, blocked P-selectin upregulation and neutrophil aggregation to baseline. Complete abrogation of platelet- neutrophil adhesion required both ZK36374 and anti-CD18. Electron microscopic observations of fixed blood specimens revealed that platelets augmented neutrophil aggregation both by forming bridges between neutrophils and through contact-mediated activation. Conclusions - The results are consistent with a model in which venous levels of shear support platelet adherence to neutrophils via P-selectin binding PSGL-1. This interaction alone is sufficient to mediate neutrophil aggregation. Abrogation of platelet adhesion and aggregation requires blocking Mac-1 in addition to PSGL-1 or P-selectin. The described mechanisms are likely of key importance in the pathogenesis and progression of thrombotic disorders that are exacerbated by leukocyte- platelet aggregation.

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