Please wait a minute...
Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2016, Vol. 10 Issue (1) : 76-84     DOI: 10.1007/s11684-015-0426-7
RESEARCH ARTICLE |
Anti-β2 glycoprotein I antibodies in complex with β2 glycoprotein I induce platelet activation via two receptors: apolipoprotein E receptor 2' and glycoprotein I bα
Wenjing Zhang,Fei Gao,Donghe Lu,Na Sun,Xiaoxue Yin,Meili Jin,Yanhong Liu()
Department of Laboratory Diagnosis, the Second Affiliated Hospital of Harbin Medical University, Harbin 150086, China
Download: PDF(1078 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract  

Anti-β2 glycoprotein I (anti-β2GP I ) antibodies are important contributors to thrombosis, especially in patients with antiphospholipid syndrome (APS). However, the mechanism by which anti-β2GP I antibodies are involved in the pathogenesis of thrombosis is not fully understood. In this report, we investigated the role of anti-β2GP I antibodies in complexes with β2GP I as mediators of platelet activation, which can serve as a potential source contributing to thrombosis. We examined the involvement of the apolipoprotein E receptor 2' (apoER2') and glycoprotein I ba (GP I bα) in platelet activation induced by the anti-β2GP I /β2GP I complex. The interaction between the anti-β2GP I /β2GP I complex and platelets was examined using in vitro methods, in which the Fc portion of the antibody was immobilized using protein A coated onto a microtiter plate. Platelet activation was assessed by measuring GP II b/ III a activation and P-selectin expression and thromboxane B2 production as well as p38 mitogen-activated protein kinase phosphorylation. Our results revealed that the anti-β2GP I /β2GP I complex was able to activate platelets, and this activation was inhibited by either the anti-GP I bα antibody or the apoER2' inhibitor. Results showed that the anti-β2GP I /β2GP I complex induced platelet activation via GP I bα and apoER2', which may then contribute to the prothrombotic tendency in APS patients.

Keywords anti-β2GP I /β2GP I complex      platelet      GP I bα      apoER2'      thrombosis     
Corresponding Authors: Yanhong Liu   
Just Accepted Date: 09 November 2015   Online First Date: 01 December 2015    Issue Date: 31 March 2016
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-015-0426-7     OR     http://academic.hep.com.cn/fmd/EN/Y2016/V10/I1/76
Fig.1  Specific of b2GP I and anti-b2GP I antibodies, and the anti-b2GP I antibodies and b2GP I binding assay. (A) b2GP I (Sino Biological Inc.) or purified b2GP I were subjected to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and incubated in the primary antibodies against b2GP I. (B) b2GP I (Sino Biological Inc.) was subjected to SDS-PAGE and incubated in the primary antibodies against b2GP I (Sino Biological Inc.) or purified anti-b2GP I antibodies. b2GP I was then incubated in HRP-conjugated goat anti-mouse. (C) Different concentrations of dilution anti-b2GP I antibodies (2, 5, 10, 20, 50, and 100 mg/ml) interacted with b2GP I (100 mg/ml), which was immobilized to plates. The data are expressed as mean±SD. n = 10; *P<0.05 versus the other groups.
Fig.2  The anti-b2GP I/b2GP I complex induced CD62P expression and GP II b/III a activation as well as TXB2 production of platelet. Human platelets were treated with PBS, the IgG/b2GP I complex (10/100 mg/ml), the anti-b2GP I/BSA complex (10/100 mg/ml), or the anti-b2GP I/b2GP I complex (10/100 mg/ml). (A) The cells were imaged via flow cytometry. (B) TXB2 production in human platelets was monitored using ELISA text. The data are expressed as mean±SD. In A and B, n = 10; *P<0.05 versus the other treatment.
Fig.3  CD62P expression and GP II b/III a activation, and TXB2 production of platelet inhibited by AK2 and RAP. Human platelets were treated with PBS, thrombin (20 mmol/L), anti-b2GP I /b2GP I complex, anti-b2GP I/b2GP I complex and RAP (0.45 mmol/L), anti-b2GP I/b2GP I complex and AK2 (0.2 mmol/L), or anti-b2GP I/b2GP I complex and RAP and AK2. (A) The cells were imaged via flow cytometry. (B) TXB2 production in human platelets was monitored using ELISA text. The data are expressed as mean±SD. In A and B, n = 10; *P<0.05 versus the IgG treatment; **P<0.05 versus the anti-b2GP I/b2GP I complex treatment.
Fig.4  The anti-b2GP I/b2GP I complex induced p38MAPK phosphorylation and TXB2 production of platelets. (A) Ratio of P-p38MAPK to total p38MAPK in platelets after treatment with the anti-b2GP I/b2GP Icomplex (10/100 mg/ml). A slight increase in P-p38MAPK expression from 5 min to 40 min, which was significant at 30 min. (B) Ratio of P-p38MAPK to total p38MAPK in washed human platelets after stimulation with PBS, IgG/b2GP I complex, anti-b2GP I/BSA complex, or anti-b2GP I/b2GP Icomplex for 30 min. (C) Ratio of P-p38MAPK to total p38MAPK in human platelets after treatment with PBS, thrombin (20 mmol/L), anti-b2GP I/b2GP I complex (10/100 mg/ml), or anti-b2GP I/b2GP I complex and SB203580 (20 mmol/L) for 30 min. (D) TXB2 expression in human platelets after injection with PBS, IgG/b2GP I complex, anti-b2GP I/BSA complex, or anti-b2GP I/b2GP I complex. The data are expressed as mean±SD. In A and B, n = 6; *P<0.05 versus the other treatment. In C and D, n = 6; *P<0.05 versus the PBS treatment; **P<0.05 versus the anti-b2GP I/b2GP I complex treatment.
Fig.5  AK2 and RAP reduced the p38MAPK phosphorylation in platelets. Ratio of P-p38MAPK to total p38MAPK in human platelets after stimulation with PBS, thrombin (20 mmol/L), anti-b2GP I/b2GP I complex, anti-b2GP I/b2GP I complex and RAP, anti-b2GP I/b2GP I complex and AK2, or anti-b2GP I/b2GP I complex and RAP and AK2. The data are expressed as mean±SD. n = 6; *P<0.05 versus IgG treatment; **P<0.05 versus anti-b2GP I/b2GP I complex treatment.
Fig.6  The anti-b2GP I/b2GP I complex induces platelet activation via apoER2' and GP Ibα. When one anti-b2GP I antibody interacts with two molecules of b2GP I, the resulting anti-b2GP I/b2GP I complex increases the affinity for phospholipids on the outer surface of the platelet membrane, which then allows for a coalescence with apoER2' and GP Ibα due to the effects of mass action. The interaction between the anti-b2GP I/b2GP I complex binding to apoER2' and GP Ibα then activates the p38MAPK signaling pathway. The initiation of this p38MAPK pathway promotes GP II b/III a activation and P-selectin expression as well as TXB2 production, which subsequently induces platelet activation either through direct or indirect mechanisms.
1 Atsumi T, Khamashta MA, Haworth RS, Brooks G, Amengual O, Ichikawa K, Koike T, Hughes GR. Arterial disease and thrombosis in the antiphospholipid syndrome: a pathogenic role for endothelin 1. Arthritis Rheum 1998; 41(5): 800–807
doi: 10.1002/1529-0131(199805)41:5<800::AID-ART5>3.0.CO;2-J pmid: 9588730
2 Galli M, Luciani D, Bertolini G, Barbui T. Anti-β 2-glycoprotein I, antiprothrombin antibodies, and the risk of thrombosis in the antiphospholipid syndrome. Blood 2003; 102(8): 2717–2723
doi: 10.1182/blood-2002-11-3334 pmid: 12816875
3 de Laat HB, Derksen RH, Urbanus RT, Roest M, de Groot PG. β2-glycoprotein I-dependent lupus anticoagulant highly correlates with thrombosis in the antiphospholipid syndrome. Blood 2004; 104(12): 3598–3602
doi: 10.1182/blood-2004-03-1107 pmid: 15315975
4 Long BR, Leya F. The role of antiphospholipid syndrome in cardiovascular disease. Hematol Oncol Clin North Am 2008; 22(1): 79–94, vi–vii
doi: 10.1016/j.hoc.2007.10.002 pmid: 18207067
5 Lozier J, Takahashi N, Putnam FW. Complete amino acid sequence of human plasma β 2-glycoprotein I. Proc Natl Acad Sci USA 1984; 81(12): 3640–3644
doi: 10.1073/pnas.81.12.3640 pmid: 6587378
6 Willems GM, Janssen MP, Pelsers MM, Comfurius P, Galli M, Zwaal RF, Bevers EM. Role of divalency in the high-affinity binding of anticardiolipin antibody-β 2-glycoprotein I complexes to lipid membranes. Biochemistry 1996; 35(43): 13833–13842
doi: 10.1021/bi960657q pmid: 8901526
7 Sheng Y, Sali A, Herzog H, Lahnstein J, Krilis SA. Site-directed mutagenesis of recombinant human β 2-glycoprotein I identifies a cluster of lysine residues that are critical for phospholipid binding and anti-cardiolipin antibody activity. J Immunol 1996; 157(8): 3744–3751
pmid: 8871678
8 Shi T, Giannakopoulos B, Yan X, Yu P, Berndt MC, Andrews RK, Rivera J, Iverson GM, Cockerill KA, Linnik MD, Krilis SA. Anti-b2-glycoprotein I antibodies in complex with b2-glycoprotein I can activate platelets in a dysregulated manner via glycoprotein Ib-IX-V. Arthritis Rheum 2006; 54(8): 2558–2567
doi: 10.1002/art.21968 pmid: 16868978
9 van Lummel M, Pennings MTT, Derksen RHWM, Urbanus RT, Lutters BC, Kaldenhoven N, de Groot PG. The binding site in β2-glycoprotein I for ApoER2′ on platelets is located in domain V. J Biol Chem 2005; 280(44): 36729–36736
doi: 10.1074/jbc.M504172200 pmid: 16091370
10 Agar C, van Os GM, Mörgelin M, Sprenger RR, Marquart JA, Urbanus RT, Derksen RH, Meijers JC, de Groot PG. β2-glycoprotein I can exist in 2 conformations: implications for our understanding of the antiphospholipid syndrome. Blood. 2010; 116(8):1336–1343 PMID: 20462962
11 Urbanus RT, Siegerink B, Roest M, Rosendaal FR, de Groot PG, Algra A. Antiphospholipid antibodies and risk of myocardial infarction and ischaemic stroke in young women in the RATIO study: a case-control study. Lancet Neurol 2009; 8(11): 998–1005
doi: 10.1016/S1474-4422(09)70239-X pmid: 19783216
12 Lutters BCH, Meijers JCM, Derksen RHWM, Arnout J, de Groot PG.Dimers of β 2-glycoprotein I mimic the in vitro effects of β 2-glycoprotein I-anti-β 2-glycoprotein I antibody complexes. J Biol Chem 2001; 276(5): 3060–3067
doi: 10.1074/jbc.M008224200 pmid: 11053420
13 Sheng Y, Kandiah DA, Krilis SA. Anti-β 2-glycoprotein I autoantibodies from patients with the “antiphospholipid” syndrome bind to β 2-glycoprotein I with low affinity: dimerization of β 2-glycoprotein I induces a significant increase in anti-β 2-glycoprotein I antibody affinity. J Immunol 1998; 161(4): 2038–2043
pmid: 9712077
14 Xie H, Zhou H, Wang H, Chen D, Xia L, Wang T, Yan J. Anti-b2GP I/b2GP I-induced TF and TNF-α expression in monocytes involving both TLR4/MyD88 and TLR4/TRIF signaling pathways. Mol Immunol 2013; 53(3): 246–254
doi: 10.1016/j.molimm.2012.08.012 pmid: 22964479
15 Zhou H, Chen D, Xie H, Xia L, Wang T, Yuan W, Yan J. Activation of MAPKs in the anti-b2GP I/b2GP I-induced tissue factor expression through TLR4/IRAKs pathway in THP-1 cells. Thromb Res 2012; 130(4): 229–235
doi: 10.1016/j.thromres.2012.08.303
16 Urbanus RT, Pennings MT, Derksen RH, de Groot PG. Platelet activation by dimeric b2-glycoprotein I requires signaling via both glycoprotein Ibα and apolipoprotein E receptor 2′. J Thromb Haemost 2008; 6(8): 1405–1412
doi: 10.1111/j.1538-7836.2008.03021.x pmid: 18485085
17 Arthur JF, Gardiner EE, Matzaris M, Taylor SG, Wijeyewickrema L, Ozaki Y, Kahn ML, Andrews RK, Berndt MC. Glycoprotein VI is associated with GP Ib-IX-V on the membrane of resting and activated platelets. Thromb Haemost 2005; 93(4): 716–723
pmid: 15841318
18 Sun B, Li J, Kambayashi J. Interaction between GP Ibα and FcγIIA receptor in human platelets. Biochem Biophys Res Commun 1999; 266(1): 24–27
doi: 10.1006/bbrc.1999.1761 pmid: 10581159
19 Furman MI, Nurden P, Berndt MC, Nurden AT, Benoit SE, Barnard MR, Ofosu FA, Michelson AD. The cleaved peptide of PAR1 results in a redistribution of the platelet surface GP Ib-IX-V complex to the surface-connected canalicular system. Thromb Haemost 2000; 84(5): 897–903
pmid: 11127874
20 Pennings MT, Derksen RH, van Lummel M, Adelmeijer J, VanHoorelbeke K, Urbanus RT, Lisman T, de Groot PG. Platelet adhesion to dimeric b-glycoprotein I under conditions of flow is mediated by at least two receptors: glycoprotein Ibα and apolipoprotein E receptor 2′. J Thromb Haemost 2007; 5(2): 369–377
doi: 10.1111/j.1538-7836.2007.02310.x pmid: 17096706
21 Lambrianides A, Carroll CJ, Pierangeli SS, Pericleous C, Branch W, Rice J, Latchman DS, Townsend P, Isenberg DA, Rahman A, Giles IP. Effects of polyclonal IgG derived from patients with different clinical types of the antiphospholipid syndrome on monocyte signaling pathways. J Immunol 2010; 184(12): 6622–6628
doi: 10.4049/jimmunol.0902765 pmid: 20483743
[1] Guocheng LIU MD, Shouhua YANG MD, Zehua WANG MD, . The relationship between platelet-derived growth factor expression and angiogenesis/lymphangiogenesis in cervical cancer[J]. Front. Med., 2009, 3(4): 447-451.
[2] Rui ZHU, Lin SHEN. Expression of protease-activated receptors on platelets in healthy individuals[J]. Front Med Chin, 2009, 3(2): 236-239.
[3] XIA Dasheng, SONG Yanqiu, LI Chao, ZHANG Feng, WEI Minxin. The change of serum leptin and its relationship with platelet membrane glycoprotein Ib in patients with coronary heart disease[J]. Front. Med., 2007, 1(4): 352-355.
[4] GONG Xiaowei, WEI Jie, LI Yusheng, CHENG Weiwei, DENG Peng, JIANG Yong. Involvement of p38 mitogen-activated protein kinase in the regulation of platelet-derived growth factor -induced cell migration[J]. Front. Med., 2007, 1(3): 248-252.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed