Human monoclonal antiphospholipid antibodies disrupt the annexin A5 anticoagulant crystal shield on phospholipid bilayers: evidence from atomic force microscopy and functional assay.
The antiphospholipid (aPL) syndrome is an autoimmune condition that is marked by recurrent pregnancy losses and/or systemic vascular thrombosis in patients who have antibodies against phospholipid/co-factor complexes. The mechanism(s) for pregnancy losses and thrombosis in this condition is (are) not known. Annexin A5 is a potent anticoagulant protein, expressed by placental trophoblasts and endothelial cells, that crystallizes over anionic phospholipids, shielding them from availability for coagulation reactions. We previously presented data supporting the hypothesis that aPL antibody-mediated disruption of the anticoagulant annexin A5 shield could be a thrombogenic mechanism in the aPL syndrome. However, this has remained a subject of controversy. We therefore used atomic force microscopy, a method previously used to study the crystallization of annexin A5, to image the effects of monoclonal human aPL antibodies on the crystal structure of the protein over phospholipid bilayers. In the presence of the aPL monoclonal antibodies (mAbs) and beta(2)-GPI, the major aPL co-factor, structures presumed to be aPL mAb-antigen complexes were associated with varying degrees of disruption to the annexin A5 crystallization pattern over the bilayer. In addition, measurements of prothrombinase activity on the phospholipid bilayers showed that the aPL mAbs reduced the anti-coagulant effect of annexin A5 and promoted thrombin generation. These data provide morphological evidence that support the hypothesis that aPL antibodies can disrupt annexin A5 binding to phospholipid membranes and permit increased generation of thrombin. The aPL antibody-mediated disruption of the annexin A5 anticoagulant shield may be an important prothrombotic mechanism in the aPL syndrome.
Rand
Jacob H
JH
Department of Pathology, Montefiore Medical Center, Albert Einstein College of Medicine, Moses Division Campus, Core Laboratory Office, North 8, 110 East 210th Street, Bronx, NY 10467, USA. jrand@montefiore.org
Wu
Xiao-Xuan
XX
Quinn
Anthony S
AS
Chen
Pojen P
PP
McCrae
Keith R
KR
Bovill
Edwin G
EG
Taatjes
Douglas J
DJ
D017304:Annexin A5 / Q000737:chemistry* / Q000187:drug effects*; D017152:Antibodies, Antiphospholipid / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000925:Anticoagulants / Q000737:chemistry*; D003460:Crystallization; D004789:Enzyme Activation; D006801:Humans; D008051:Lipid Bilayers; D018625:Microscopy, Atomic Force; D010743:Phospholipids; D013925:Thromboplastin / Q000378:metabolism
D017304:Annexin A5; D017152:Antibodies, Antiphospholipid; D000911:Antibodies, Monoclonal; D000925:Anticoagulants; D008051:Lipid Bilayers; D010743:Phospholipids; D013925:Thromboplastin