Antiphospholipid antibodies promote leukocyte-endothelial cell adhesion and thrombosis in mice by antagonizing eNOS via β2GPI and apoER2

J Clin Invest. 2011 Jan;121(1):120-31. doi: 10.1172/JCI39828. Epub 2010 Dec 1.


In antiphospholipid syndrome (APS), antiphospholipid antibodies (aPL) binding to β2 glycoprotein I (β2GPI) induce endothelial cell-leukocyte adhesion and thrombus formation via unknown mechanisms. Here we show that in mice both of these processes are caused by the inhibition of eNOS. In studies of cultured human, bovine, and mouse endothelial cells, the promotion of monocyte adhesion by aPL entailed decreased bioavailable NO, and aPL fully antagonized eNOS activation by diverse agonists. Similarly, NO-dependent, acetylcholine-induced increases in carotid vascular conductance were impaired in aPL-treated mice. The inhibition of eNOS was caused by antibody recognition of domain I of β2GPI and β2GPI dimerization, and it was due to attenuated eNOS S1179 phosphorylation mediated by protein phosphatase 2A (PP2A). Furthermore, LDL receptor family member antagonism with receptor-associated protein (RAP) prevented aPL inhibition of eNOS in cell culture, and ApoER2-/- mice were protected from aPL inhibition of eNOS in vivo. Moreover, both aPL-induced increases in leukocyte-endothelial cell adhesion and thrombus formation were absent in eNOS-/- and in ApoER2-/- mice. Thus, aPL-induced leukocyte-endothelial cell adhesion and thrombosis are caused by eNOS antagonism, which is due to impaired S1179 phosphorylation mediated by β2GPI, apoER2, and PP2A. Our results suggest that novel therapies for APS can now be developed targeting these mechanisms.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Antiphospholipid / metabolism*
  • Cattle
  • Cell Adhesion / immunology
  • Cell Adhesion / physiology
  • Dimerization
  • Endothelial Cells / immunology*
  • Endothelial Cells / metabolism
  • Humans
  • In Vitro Techniques
  • LDL-Receptor Related Proteins / antagonists & inhibitors
  • LDL-Receptor Related Proteins / deficiency
  • LDL-Receptor Related Proteins / immunology
  • Leukocytes / immunology*
  • Leukocytes / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Models, Biological
  • Nitric Oxide / physiology
  • Nitric Oxide Synthase Type III / antagonists & inhibitors
  • Nitric Oxide Synthase Type III / deficiency
  • Nitric Oxide Synthase Type III / immunology
  • Protein Phosphatase 2 / antagonists & inhibitors
  • Protein Phosphatase 2 / genetics
  • Protein Phosphatase 2 / metabolism
  • Protein Structure, Tertiary
  • RNA, Small Interfering / genetics
  • Thrombosis / etiology
  • Thrombosis / immunology*
  • beta 2-Glycoprotein I / antagonists & inhibitors
  • beta 2-Glycoprotein I / chemistry
  • beta 2-Glycoprotein I / immunology


  • Antibodies, Antiphospholipid
  • LDL-Receptor Related Proteins
  • RNA, Small Interfering
  • beta 2-Glycoprotein I
  • low density lipoprotein receptor-related protein 8
  • Nitric Oxide
  • Nitric Oxide Synthase Type III
  • Nos3 protein, mouse
  • Protein Phosphatase 2