In vitro effect of anti-β₂ glycoprotein I antibodies on P-selectin expression, a marker of platelet activation

Reumatismo. 2012 Mar 19;64(1):35-9. doi: 10.4081/reumatismo.2012.35.

Abstract

Objective: Antiphospholipid antibodies (aPL) associated with thrombembolic events and/or pregnancy morbidity characterize the so-called antiphospholipid syndrome (APS). Beta2glycoprotein I (β2GPI) represents the major target antigen for aPL, but the pathogenic role of anti-β2GPI antibodies (aβ2GPI) is still unclear. Some authors assume they play a role in activating platelets. The effects of aβ2GPI antibodies on platelet P-selectin expression were evaluated in this study.

Methods: Aβ2GPI antibodies in the plasma of a pregnant APS patient were isolated by affinity chromatography during two different stages (catastrophic and quiescent) of the disease. Gel filtered platelets (100,000/μl) from healthy volunteers were incubated with β2-GPI (20 μg/ml) and with different concentrations (5, 25 e 50 μg/ml) of aβ2GPI antibodies. P-selectin surface expression on platelets was assessed by flow cytometry using a specific fluorescent antibody directed against P-selectin.

Results: Aβ2GPI antibodies induced platelet activation only in the presence of thrombin receptor activator for peptide 6 (TRAP-6), a platelet agonist, at a subthreshold concentration. Aβ2GPI antibody enhancement on platelet surface P-selectin expression was stronger in the catastrophic than in the quiescent phase of the disease (47% versus 15%).

Conclusions: TRAP-6 dependent platelet activation by aβ2GPI antibodies is consistent with the “two hit” pathogenetic hypothesis for thrombosis. Aβ2GPI antibodies induce higher platelet P-selectin expression during the active rather than in the acute phases.

Publication types

  • Comparative Study

MeSH terms

  • Acute Disease
  • Adult
  • Antiphospholipid Syndrome / blood*
  • Antiphospholipid Syndrome / complications
  • Antiphospholipid Syndrome / immunology
  • Autoantibodies / immunology
  • Autoantibodies / isolation & purification
  • Autoantibodies / pharmacology*
  • Autoantigens / immunology*
  • Autoantigens / isolation & purification
  • Chromatography, Affinity
  • Female
  • Flow Cytometry
  • Humans
  • In Vitro Techniques
  • P-Selectin / biosynthesis*
  • P-Selectin / genetics
  • Peptide Fragments / pharmacology
  • Platelet Activation*
  • Pregnancy
  • Pregnancy Complications / blood*
  • Pregnancy Complications / immunology
  • Pregnancy Complications, Hematologic / blood
  • Pregnancy Complications, Hematologic / etiology
  • Pregnancy Complications, Hematologic / immunology
  • Thrombophilia / blood
  • Thrombophilia / etiology*
  • Thrombophilia / immunology
  • beta 2-Glycoprotein I / immunology*
  • beta 2-Glycoprotein I / isolation & purification
  • beta 2-Glycoprotein I / pharmacology

Substances

  • Autoantibodies
  • Autoantigens
  • P-Selectin
  • Peptide Fragments
  • beta 2-Glycoprotein I
  • thrombin receptor peptide (42-47)