Thromb Haemost 2019; 119(07): 1147-1153
DOI: 10.1055/s-0039-1685453
Blood Cells, Inflammation and Infection
Georg Thieme Verlag KG Stuttgart · New York

Platelet Activation by Antiphospholipid Antibodies Depends on Epitope Specificity and is Prevented by mTOR Inhibitors

Anne Hollerbach
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
2   Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
,
Nadine Müller-Calleja
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
2   Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
,
Svenja Ritter
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
,
Friederike Häuser
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
,
Antje Canisius
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
,
Carolin Orning
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
,
Kerstin Jurk
2   Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
,
Karl J. Lackner
1   Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
› Institutsangaben
Funding This study was supported by the German Federal Ministry of Education and Research (BMBF 01EO1003). The authors are responsible for the contents of this publication.
Weitere Informationen

Publikationsverlauf

06. September 2018

03. März 2019

Publikationsdatum:
24. April 2019 (online)

Abstract

Antiphospholipid antibodies (aPL) have been reported to activate platelets. This is considered to be one of the pathogenic properties of aPL. Even though aPL heterogeneity is quite well established, little is known, if the ability to activate platelets is common to all aPL or depends on antigen specificity. To further study this issue, we analyzed the ability of three human monoclonal aPL with distinctly different antigenic specificities to activate platelets in vitro. The results obtained with human monoclonal aPL were validated with immunoglobulin G (IgG) fractions obtained from patients with antiphospholipid syndrome (APS). A co-factor-independent human monoclonal anticardiolipin aPL had no discernible effect on human platelets. Two monoclonal aPL reactive against β2 glycoprotein I (β2GPI) induced platelet aggregation, integrin αIIbβ3 activation and P-selectin surface expression. These data could be confirmed with patient IgG fractions which could only induce aggregation, if they had anti-β2GPI activity. Anti-β2GPI aPL-induced platelet activation depended on interaction of aPL with the low affinity Fcγ-receptor IIa on the platelet surface. It was completely abolished by pretreatment of platelet-rich plasma with the mechanistic target of rapamycin (mTOR) inhibitors rapamycin or everolimus. This extends previous data showing that mTOR is involved in signaling of anti-β2GPI in monocytes and endothelial cells. In conclusion, anti-β2GPI aPL activate platelets while co-factor-independent anticardiolipin aPL have no effect. mTOR is involved in this signaling process which has implications beyond APS, because so far the role of mTOR signaling in platelets is incompletely explored and requires further study.

Authors' Contributions

A.H. performed experiments and wrote the manuscript; N.M.-C. designed the study, performed experiments and wrote the manuscript; S.R., F.H., A.C. and C.O. performed experiments; K.J. provided expert technical advice and revised the manuscript; and K.J.L. designed the study and wrote the manuscript.


Supplementary Material

 
  • References

  • 1 Meroni PL, Borghi MO, Raschi E, Tedesco F. Pathogenesis of antiphospholipid syndrome: understanding the antibodies. Nat Rev Rheumatol 2011; 7 (06) 330-339
  • 2 Willis R, Harris EN, Pierangeli SS. Pathogenesis of the antiphospholipid syndrome. Semin Thromb Hemost 2012; 38 (04) 305-321
  • 3 Giannakopoulos B, Krilis SA. The pathogenesis of the antiphospholipid syndrome. N Engl J Med 2013; 368 (11) 1033-1044
  • 4 de Groot PG, Urbanus RT. The significance of autoantibodies against β2-glycoprotein I. Blood 2012; 120 (02) 266-274
  • 5 Müller-Calleja N, Lackner KJ. Mechanisms of cellular activation in the antiphospholipid syndrome. Semin Thromb Hemost 2018; 44 (05) 483-492
  • 6 Müller-Calleja N, Hollerbach A, Häuser F, Canisius A, Orning C, Lackner KJ. Antiphospholipid antibody-induced cellular responses depend on epitope specificity : implications for treatment of antiphospholipid syndrome. J Thromb Haemost 2017; 15 (12) 2367-2376
  • 7 Urbanus RT, Derksen RH, de Groot PG. Platelets and the antiphospholipid syndrome. Lupus 2008; 17 (10) 888-894
  • 8 Baroni G, Banzato A, Bison E, Denas G, Zoppellaro G, Pengo V. The role of platelets in antiphospholipid syndrome. Platelets 2017; 28 (08) 762-766
  • 9 von Landenberg C, Lackner KJ, von Landenberg P, Lang B, Schmitz G. Isolation and characterization of two human monoclonal anti-phospholipid IgG from patients with autoimmune disease. J Autoimmun 1999; 13 (02) 215-223
  • 10 Buschmann C, Fischer C, Ochsenhirt V, Neukirch C, Lackner KJ, von Landenberg P. Generation and characterization of three monoclonal IgM antiphospholipid antibodies recognizing different phospholipid antigens. Ann N Y Acad Sci 2005; 1051: 240-254
  • 11 Prinz N, Häuser F, Lorenz M, Lackner KJ, von Landenberg P. Structural and functional characterization of a human IgG monoclonal antiphospholipid antibody. Immunobiology 2011; 216 (1-2): 145-151
  • 12 Müller-Calleja N, Ritter S, Hollerbach A, Falter T, Lackner KJ, Ruf W. Complement C5 but not C3 is expendable for tissue factor activation by cofactor-independent antiphospholipid antibodies. Blood Adv 2018; 2 (09) 979-986
  • 13 Manukyan D, Müller-Calleja N, Jäckel S. , et al. Cofactor-independent human antiphospholipid antibodies induce venous thrombosis in mice. J Thromb Haemost 2016; 14 (05) 1011-1020
  • 14 Lackner KJ, von Landenberg C, Barlage S, Schmitz G. Analysis of prothrombotic effects of two human monoclonal IgG antiphospholipid antibodies of apparently similar specificity. Thromb Haemost 2000; 83 (04) 583-588
  • 15 Campbell AL, Pierangeli SS, Wellhausen S, Harris EN. Comparison of the effects of anticardiolipin antibodies from patients with the antiphospholipid syndrome and with syphilis on platelet activation and aggregation. Thromb Haemost 1995; 73 (03) 529-534
  • 16 Wiener MH, Burke M, Fried M, Yust I. Thromboagglutination by anticardiolipin antibody complex in the antiphospholipid syndrome: a possible mechanism of immune-mediated thrombosis. Thromb Res 2001; 103 (03) 193-199
  • 17 Urbanus RT, Pennings MTT, Derksen RHWM, de Groot PG. Platelet activation by dimeric β2-glycoprotein I requires signaling via both glycoprotein Ibalpha and apolipoprotein E receptor 2′. J Thromb Haemost 2008; 6 (08) 1405-1412
  • 18 Canaud G, Bienaimé F, Tabarin F. , et al. Inhibition of the mTORC pathway in the antiphospholipid syndrome. N Engl J Med 2014; 371 (04) 303-312
  • 19 Babinska A, Markell MS, Salifu MO, Akoad M, Ehrlich YH, Kornecki E. Enhancement of human platelet aggregation and secretion induced by rapamycin. Nephrol Dial Transplant 1998; 13 (12) 3153-3159
  • 20 Wu Q, Huang KS, Chen M, Huang DJ. Rapamycin enhances platelet aggregation induced by adenosine diphosphate in vitro. Platelets 2009; 20 (06) 428-431
  • 21 Graff J, Harder S, Reiche C, Scheuermann EH, Gossmann J. Platelet activation markers after conversion from azathioprine to sirolimus based immunosuppression in renal transplant recipients. Int J Clin Pharmacol Ther 2012; 50 (12) 873-879
  • 22 Aslan JE, Tormoen GW, Loren CP, Pang J, McCarty OJ. S6K1 and mTOR regulate Rac1-driven platelet activation and aggregation. Blood 2011; 118 (11) 3129-3136
  • 23 Cao H, Bissinger R, Umbach AT, Gawaz M, Lang F. Temsirolimus sensitive stimulation of platelet activity, apoptosis and aggregation by collagen related peptide. Cell Physiol Biochem 2017; 42 (03) 1252-1263
  • 24 Liu Y, Hu M, Luo D. , et al. Class III PI3K positively regulates platelet activation and thrombosis via PI(3)P-directed function of NADPH oxidase. Arterioscler Thromb Vasc Biol 2017; 37 (11) 2075-2086
  • 25 Arnout J. The pathogenesis of the antiphospholipid syndrome: a hypothesis based on parallelisms with heparin-induced thrombocytopenia. Thromb Haemost 1996; 75 (04) 536-541
  • 26 Arvieux J, Roussel B, Pouzol P, Colomb MG. Platelet activating properties of murine monoclonal antibodies to beta 2-glycoprotein I. Thromb Haemost 1993; 70 (02) 336-341
  • 27 Lutters BC, Derksen RH, Tekelenburg WL, Lenting PJ, Arnout J, de Groot PG. Dimers of beta 2-glycoprotein I increase platelet deposition to collagen via interaction with phospholipids and the apolipoprotein E receptor 2′. J Biol Chem 2003; 278 (36) 33831-33838
  • 28 Pengo V, Ruffatti A, Legnani C. , et al. Clinical course of high-risk patients diagnosed with antiphospholipid syndrome. J Thromb Haemost 2010; 8 (02) 237-242
  • 29 Pengo V, Ruffatti A, Legnani C. , et al. Incidence of a first thromboembolic event in asymptomatic carriers of high-risk antiphospholipid antibody profile: a multicenter prospective study. Blood 2011; 118 (17) 4714-4718