Hydroxychloroquine Reverses Platelet Activation Induced by Human IgG Antiphospholipid Antibodies

Ricardo G. Espinola; Silvia S. Pierangeli; Azzudin E. Ghara; E. Nigel Harris

doi:10.1055/s-0037-1613033

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 87(03): 518-522
DOI: 10.1055/s-0037-1613033

Review Article

Schattauer GmbH

Hydroxychloroquine Reverses Platelet Activation Induced by Human IgG Antiphospholipid Antibodies

Ricardo G. Espinola

¹Department of Microbiology, Biochemistry and Immunology, and Medicine

,

Silvia S. Pierangeli

¹Department of Microbiology, Biochemistry and Immunology, and Medicine

,

Azzudin E. Ghara

²Morehouse School of Medicine, Atlanta, Georgia, USA

,

E. Nigel Harris

²Morehouse School of Medicine, Atlanta, Georgia, USA

› Author Affiliations

Abstract

Summary

Prothrombotic properties of antiphospholipid (aPL) antibodies may be explained in part by their ability to enhance the activation of platelets pre-treated with low doses of ADP or thrombin. The antimalarial drug hydroxychloroquine (HQ) has been used successfully in prevention of postoperative thrombosis and in treatment of patients with SLE or APS. In one study, administration of HQ reversed the thrombogenic properties of aPL in mice. However, the mechanism of action of HQ in preventing thrombosis is not clearly understood. In order to explore this further, the effects of HQ on activation of platelets by aPL in the presence of a thrombin agonist was studied. The changes in the expression of GPIIb/IIIa (CD41a) and GPIIIa (CD61) on platelet membrane by flow cytometry were used as indicators of platelet activation. Citrated whole blood from a healthy donor was treated at room temperature with suboptimal doses of a thrombin agonist receptor peptide (TRAP) and affinity-purified aPL antibodies, in the presence and in the absence of hydroxychloroquine (1 mM). TRAP increased the expression of GPIIb/IIIa and GPIIIa on platelet surface. The treatment of the platelets with the six aPL antibodies in the presence of 12 nMol/ml TRAP further increased the expression of GPIIb/IIIa by 42.3 ± 12.3% and the expression of GPIIIa was further incremented by 46.8 ± 13.5%. The effects of aPL and TRAP on expression of platelet surface markers of activation was completely abrogated by HQ in a dose-dependent fashion and was effective at concentrations of HQ as low as 25 µg/ml (0.0125 mM). This suggests at least one possible mechanism by which HQ may prevent thrombosis. This may have important implications in treatment of thrombosis in APS patients.

Keywords

Antiphospholipid antibodies - platelet activation - hydroxychloroquine - thrombosis - GPIIb/IIIa

Full Text

References

References
1 Harris EN. Syndrome of the Black Swan. Br J Rheumatol 1987; 26: 324-6.
2 Khamashta MA, Harris EN, Gharavi AE. Immune mediated mechanism for thrombosis: antiphospholipid antibody binding to platelet membranes. Ann Rheum Dis 1988; 47: 849-54.
3 Campbell AL, Pierangeli SS, Wellhausen S, Harris EN. Comparison of the effect of anticardiolipin antibodies from patients with the antiphospholipid syndrome and with syphilis on platelet activation and aggregation. Thromb Haemost 1995; 73: 529-34.
4 Rosove MH, Brewer P. Antiphospholipid thrombosis: clinical course after the first thrombotic event in 70 patients. Ann Intern Med 1992; 117: 303-8.
5 Derksen RHMV, de Groot PG, Kator L, Nieuwenhuis HK. Patients with antiphospholipid antibodies and venous thrombosis should receive longterm anticoagulant treatment. Ann Rheum Dis 1993; 52: 689-92.
6 Khamashta MA, Cuadrado MJ, Mujic F, Taub NA, Hunt BJ, Hughes GR. The management of thrombosis in the antiphospholipid antibody syndrome. N Engl J Med 1995; 332: 993-7.
7 Carter AE, Eban R. Prevention of postoperative deep venous thrombosis in legs by orally administered hydroxychloroquine sulphate. Br Med J 1974; 03 (923) 94-5.
8 Johnson R, Green J, Charnley J. Pulmonary embolism and its prophylaxis following the Charnley total hip replacement. Clin Orthop 1977; 127: 123-32.
9 Petri M, Hochberg M, Hellmann D, Bell W, Goldman D. Incidence of and predictors of thrombotic events in SLE: protective role of hydroxychloroquine. Arthritis Rheum 1992; 35 (09) S54.
10 Wallace D. Does hydroxychloroquine sulphate prevent clot formation in systemic lupus erythematosus?. Arthritis Rheum 1987; 30: 1435-6.
11 Edwards M, Pierangeli SS, Liu Xwei, Barker JH, Anderson GH, Harris EN. Hydroxychloroquine Reverses Thrombogenic Properties of Antiphospholipid Antibodies in Mice. Circulation 1997; 96 (12) 4380-4.
12 Bertrand E, Cloltre B, Ticolet R, Bile R, Grantier C, Abiyou G, Bohui Y. Antiaggregation action of chloroquine. Med Trop (Mars 1990; 50: 143-6.
13 Janicinova V, Nosal R, Petrokova M. On the inhibitory effect of chloroquine on blood platelet aggregation. Thromb Res 1994; 74: 495-504.
14 Pierangeli SS, Liu WeiX, Anderson GH, Barker JH, Harris EN. Induction of thrombosis in a mouse model by IgG, IgM and IgA immunoglobulins from patients with the Antiphospholipid Syndrome. Thromb Haemost 1995; 74: 1361-7.
15 Ginsberg MH, Frelinger AL, Lam SC, Forsyth J, McMillan R, Plow EF, Shattil SJ. Analysis of platelet aggregation disorders based on flow cytometric analysis of membrane glycoprotein IIb-IIIa with conformationspecific monoclonal antibodies. Blood 1990; 76 (10) 2017-23.
16 Michelson AD, Ellis PA, Barnard MR, Matic GB, Viles AF, Kestin AS. Downregulation of the platelet surface glycoprotein Ib-IX complex in whole blood stimulated by thrombin, adenosine diphosphate or an in vivo wound. Blood 1991; 77: 770-9.
17 Krnic-Barrie S, Reister CO'Connor, Looney SW, Pierangeli SS, Harris EN. A retrospective review of 61 patients with antiphospholipid syndrome. Arch Intern Med 1997; 157: 2101-8.
18 Prowse C, Pepper D, Dawes J. Prevention of the platelet alpha-granule release reaction by membrane-active drugs. Thromb Res 1982; 25 (03) 219-27.
19 Tett SE, Day RO, Cutler DJ. Concentration-effect relationship of hydroxychloroquine in rheumatoid arthritis – A Cross sectional study. J Rheumatol 1993; 20: 1874-9.
20 French JK, Hurst NP, O’Donnell ML, Betts WH. Uptake of chloroquine and hydroxychloroquine by human blood leucocytes in vitro: relation to cellular concentrations during antirheumatic therapy. Ann Rheum Dis 1987; 46: 42-5.
21 Tett S. Clinical pharmacokinetics of slow acting antirheumatic drugs. Clin Pharmacokinet 1993; 25: 392-407.
22 French J, Hurst N, O’Donnell M, Betts W. Uptake of chloroquine and hydroxychloroquine by human blood leukocytes in vitro: relation to cellular concentrations during antirheumatic therapy. Ann Rheum Dis 1987; 46: 42-5.
23 Mackenzie AH. Pharmacologic actions of 4-aminoquinoline compounds. Am J Med 1983; 75: 5-10.
24 French JK, Hurst NP, O’Donnell ML, Betss WH. Ann Rheum Dis 1987; 46: 42-5 Augustins. Chloroquine levels in blood during chronic treatment of patients with rheumatic arthritis. Eur J Clin Pharmacol 1992; 42: 429-33.
25 Hackeng CM, Relou IA, Pladet MW, Gorter G, van Rijn HJM, Akkerman J-W. Early platelet activation by low density lipoprotein via p38MAP kinase. Thromb Haemost 1999; 82: 1749-56.
26 Kuwahara M, Sugimoto M, Tsuji S, Miyata S, Yoshioka A. Cytosolic calcium changes in a process of platelet adhesion and cohesion on a von Willebrand factor-coated surface under flow conditions. Blood 1999; 94 (04) 1149-55.
27 Billah M, Lapetina E, Cuetracasas P. Phospholipase A2 activity specific for phosphatidic acid. J Biol Chem 1981; 256: 5399-403.
28 McCrea J, Robinson P, Gerrard J. Mepacrine (quinacrine) inhibition of thrombin-induced platelet responses can be overcome by lysophosphatidic acid. Biochim Biophys Acta 1985; 842: 189-94.
29 Out HJ, deGroot PG, van Vliet M, de Gast GC, Nieuwenhauis HK, Derksen RHWM. Antibodies to platelets in patients with anti-phospholipid antibodies. Blood 1991; 77: 2655-9.
30 Martinuzzo ME, Maclouf J, Carreras LO, Levy-Toledano S. Antiphospholipid antibodies enhance thrombin-induced platelet activation and thromboxane formation. Thromb Haemost 1993; 70 (04) 667-71.
31 Arvieux J, Roussel B, Pouzol P, Colomb MG. Platelet activating properties of murine monoclonal antibodies to β2-glycoprotein I. Thromb Haemost 1993; 70 (02) 336-41.