Endothelial microparticle release is stimulated in vitro by purified IgG from patients with the antiphospholipid syndrome

Charis Pericleous; Lindsey A. Clarke; Paul A. Brogan; David S. Latchman; David A. Isenberg; Yiannis Ioannou; Ian P. Giles; Anisur Rahman

doi:10.1160/TH12-05-0346

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2013; 109(01): 72-78
DOI: 10.1160/TH12-05-0346

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Endothelial microparticle release is stimulated in vitro by purified IgG from patients with the antiphospholipid syndrome

Charis Pericleous

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

,

Lindsey A. Clarke

²Institute of Child Health University College London, London, UK

,

Paul A. Brogan

²Institute of Child Health University College London, London, UK

,

David S. Latchman

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

,

David A. Isenberg

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

,

Yiannis Ioannou

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

,

Ian P. Giles

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

,

Anisur Rahman

¹Centre for Rheumatology, Division of Medicine University College London, Rayne Institute, London, UK

› Author Affiliations

Abstract

Summary

IgG antiphospholipid antibodies (aPL) exert direct effects on various cell types, contributing to the pathogenesis of thrombosis and pregnancy morbidity in patients with the antiphospholipid syndrome (APS). Some IgG samples from these patients activate endothelial cells (EC) in vitro as judged by surface expression of adhesion molecules such as E-selectin, which can promote thrombosis. Endothelial micro-particles (EMP), which themselves are potentially prothrombotic, are released by activated EC. Though elevated circulating EMP levels have been reported in patients with APS, it is not known whether these EMP are released due to a direct effect of aPL on the cells. We tested the effect of purified polyclonal IgG from patients with APS (APS-IgG) and healthy controls (HC-IgG) upon cultured human umbilical vein EC (HUVEC). HUVEC exposed to APS-IgG produced significantly more EMP than those exposed to HC-IgG (p=0.0036) and a greater proportion of these EMP carried surface E-selectin (6.2% ± 4.0 for APS-IgG vs. 3.4% ± 2.0 for HC IgG, p=0.0172). This study therefore demonstrates that purified polyclonal APS-IgG can drive EMP release. We propose that EMP generation may be a useful measure of aPL-mediated pathogenic effects upon EC.

Keywords

Antiphospholipid antibodies - endothelial cell activation - endothelial microparticles

Full Text

References

References
1 Petri M. Classification and epidemiology of the Antiphospholipid Syndrome. In: Asherson RA, Cervera R, Piette J-C, Shoenfeld Y. editors. The Antiphospholipid Syndrome II: Autoimmune Thrombosis. Second ed.. Amsterdam.: Elsevier Science B V.; 2002. pp. 11-20.
2 Rai R, Cohen H, Dave M. et al. Randomised controlled trial of aspirin and aspirin plus heparin in pregnant women with recurrent miscarriage associated with phospholipid antibodies (or antiphospholipid antibodies). Br Med J 1997; 314: 253-257. DOI:10.1136/bmj.314.7076.253.
3 Rai R, Regan L. Antiphospholipid syndrome in pregnancy: a randomized, controlled trial of treatment. Obstet Gynecol 2002; 100: 1354. DOI:10.1016/S0029-7844(02)02574-7.
4 Branch DW, Rodgers GM. Induction of endothelial cell tissue factor activity by sera from patients with antiphospholipid syndrome: a possible mechanism of thrombosis. Am J Obstet Gynecol 1993; 168: 206-210.
5 Pierangeli SS, Colden-Stanfield M, Liu X. et al. Antiphospholipid antibodies from antiphospholipid syndrome patients activate endothelial cells in vitro and in vivo. Circulation 1999; 99: 1997-2002. DOI:10.1161/01.CIR.99.15.1997.
6 Pierangeli SS, Espinola RG, Liu X. et al. Thrombogenic effects of antiphospholipid antibodies are mediated by intercellular cell adhesion molecule-1, vascular cell adhesion molecule- 1, and P-selectin. Circ Res 2001; 88: 245-250. DOI:10.1161/01.RES.88.2.245.
7 Dunoyer-Geindre S, Dimitrova Y, Fish RJ. et al. Fluvastatin increases the expression of adhesion molecules, monocyte chemoattractant protein-1 and tissue factor in HUVEC stimulated by patient IgG fractions containing antiphospholipid antibodies. Thromb Haemost 2005; 93: 339-345.
8 Satta N, Kruithof EK, Fickentscher C. et al. Toll-like receptor 2 mediates the activation of human monocytes and endothelial cells by antiphospholipid antibodies. Blood 2011; 117: 5523-5531. DOI:10.1182/blood-2010-11-316158.
9 Abid Hussein MN, Meesters EW, Osmanovic N. et al. Antigenic characterization of endothelial cell-derived microparticles and their detection ex vivo. J Thromb Haemost 2003; 01: 2434-2443. DOI:10.1046/j.1538-7836.2003.00455.x.
10 Pericleous C, Giles I, Rahman A. Are endothelial microparticles potential markers of vascular dysfunction in the antiphospholipid syndrome?. Lupus 2009; 18: 671-675. DOI:10.1177/0961203309103062.
11 Combes V, Simon AC, Grau GE. et al. In vitro generation of endothelial microparticles and possible prothrombotic activity in patients with lupus anticoagulant. J Clin Invest 1999; 104: 93-102. DOI:10.1172/JCI4985.
12 Dignat-George F, Camoin-Jau L, Sabatier F. et al. Endothelial microparticles: a potential contribution to the thrombotic complications of the antiphospholipid syndrome. Thromb Haemost 2004; 91: 667-673.
13 Jy W, Tiede M, Bidot CJ. et al. Platelet activation rather than endothelial injury identifies risk of thrombosis in subjects positive for antiphospholipid antibodies. Thromb Res 2007; 121: 319-325. DOI:10.1016/j.thromres.2007.04.014.
14 Miyakis S, Lockshin MD, Atsumi T. et al. International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemost 2006; 04: 295-306. DOI:10.1111/j.1538-7836.2006.01753.x.
15 Ioannou Y, Pericleous C, Giles I. et al. Binding of antiphospholipid antibodies to discontinuous epitopes on domain I of human beta(2)-glycoprotein I: mutation studies including residues R39 to R43. Arthritis Rheum 2007; 56: 280-290. DOI:10.1002/art.22306.
16 de Laat B, Derksen RH, Urbanus RT. et al. IgG antibodies that recognize epitope Gly40-Arg43 in domain I of beta 2-glycoprotein I cause LAC, and their presence correlates strongly with thrombosis. Blood 2005; 105: 1540-1545. DOI:10.1182/blood-2004-09-3387.
17 Ioannou Y, Romay-Penabad Z, Pericleous C. et al. In vivo inhibition of antiphospholipid antibody-induced pathogenicity utilizing the antigenic target peptide domain I of beta2-glycoprotein I: proof of concept. J Thromb Haemost 2009; 07: 833-842. DOI:10.1111/j.1538-7836.2009.03316.x.
18 Giles I, Pericleous C, Liu X. et al. Thrombin binding predicts the effects of sequence changes in a human monoclonal antiphospholipid antibody on its in vivo biologic actions. J Immunol 2009; 182: 4836-4843. DOI:10.4049/jimmunol.0804241.
19 Vega-Ostertag M, Casper K, Swerlick R. et al. Involvement of p38 MAPK in the up-regulation of tissue factor on endothelial cells by antiphospholipid antibodies. Arthritis Rheum 2005; 52: 1545-1554. DOI:10.1002/art.21009.
20 O’Reilly FM, Casper KA, Otto KB. et al. Regulation of tissue factor in microvascular dermal endothelial cells. J Invest Dermatol 2003; 120: 489-494. DOI:10.1046/j.1523-1747.2003.12063.x.
21 Simantov R, LaSala JM, Lo SK. et al. Activation of cultured vascular endothelial cells by antiphospholipid antibodies. J Clin Invest 1995; 96: 2211-2219. DOI:10.1172/JCI118276.
22 Del Papa N, Guidali L, Spatola L. et al. Relationship between anti-phospholipid and anti-endothelial cell antibodies III: beta 2 glycoprotein I mediates the antibody binding to endothelial membranes and induces the expression of adhesion molecules. Clin Exp Rheumatol 1995; 13: 179-185.
23 Del Papa N, Guidali L, Sala A. et al. Human polyclonal and monoclonal anti-beta2-glycoprotein I antibodies react in vitro with endothelial cells through adherent beta 2-glycoprotein I and induce endothelial cell activation. Arthritis Rheum 1997; 40: 551-561. DOI:10.1002/art.1780400322.
24 Pierangeli SS, Liu X, Espinola R. et al. Functional analyses of patient-derived IgG monoclonal anticardiolipin antibodies using in vivo thrombosis and in vivo microcirculation models. Thromb Haemost 2000; 84: 388-395.
25 Hamid C, Norgate K, D’Cruz DP. et al. Anti-beta2GPI-antibody-induced endothelial cell gene expression profiling reveals induction of novel pro-inflammatory genes potentially involved in primary antiphospholipid syndrome. Ann Rheum Dis 2007; 66: 1000-1007. DOI:10.1136/ard.2006.063909.
26 Vega-Ostertag ME, Ferrara DE, Romay-Penabad Z. et al. Role of p38 mitogen-activated protein kinase in antiphospholipid antibody-mediated thrombosis and endothelial cell activation. J Thromb Haemost 2007; 05: 1828-1834. DOI:10.1111/j.1538-7836.2007.02680.x.
27 Brogan PA, Shah V, Brachet C. et al. Endothelial and platelet microparticles in vasculitis of the young. Arthritis Rheum 2004; 50: 927-936. DOI:10.1002/art.20199.
28 Dunoyer-Geindre S, Boehlen F, Favier R. et al. Endothelial cell activation by immunoglobulins from patients with immune thrombocytopenic purpura or with antiphospholipid syndrome. Haematologica 2008; 93: 635-636. DOI:10.3324/haematol.12358.
29 Simoncini S, Sapet C, Camoin-Jau L. et al. Role of reactive oxygen species and p38MAPK in the induction of the pro-adhesive endothelial state mediated by IgG from patients with anti-phospholipid syndrome. International Immunity 2005; 17: 489-500. DOI:10.1093/intimm/dxh229.
30 Pierangeli SS, Harris EN. Probing antiphospholipid-mediated thrombosis: the interplay between anticardiolipin antibodies and endothelial cells. Lupus 2003; 12: 539-545. DOI:10.1191/961203303lu398oa.
31 Lopez-Pedrera C, Cuadrado MJ, Herandez V. et al. Proteomic analysis in monocytes of antiphospholipid syndrome patients: Deregulation of proteins related to the development of thrombosis. Arthritis Rheum 2008; 58: 2835-2844. DOI:10.1002/art.23756.
32 Lambrianides A, Carroll CJ, Pierangeli SS. et al. Effects of polyclonal IgG derived from patients with different clinical types of the antiphospholipid syndrome on monocyte signaling pathways. J Immunol 2010; 184: 6622-6628. DOI:10.4049/jimmunol.0902765.