Role of TLR‑4 in anti‑β2‑glycoprotein I‑induced activation of peritoneal macrophages and vascular endothelial cells in mice

Wang,Meiyun; Kong,Xiangmin; Xie,Yachao; He,Chao; Wang,Ting; Zhou,Hong

doi:10.3892/mmr.2019.10084

Role of TLR‑4 in anti‑β2‑glycoprotein I‑induced activation of peritoneal macrophages and vascular endothelial cells in mice

Authors:
- Meiyun Wang
- Xiangmin Kong
- Yachao Xie
- Chao He
- Ting Wang
- Hong Zhou
View Affiliations

Affiliations: Department of Internal Medicine, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China, Department of Clinical Laboratory and Hematology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China
Published online on: March 26, 2019 https://doi.org/10.3892/mmr.2019.10084
Pages: 4353-4363
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Anti‑phospholipid syndrome (APS) is a systematic autoimmune disease that is associated with presence of antiphospholipid antibodies (aPL), recurrent thrombosis, and fetal morbidity in pregnancy. Toll‑like receptor‑4 (TLR‑4), a member of TLR family, is known to have a fundamental role in pathogen recognition and activation of innate immunity. The β2‑glycoprotein I (β2GPI), a protein circulating in the blood at a high concentration, is able of scavenging lipopolysaccharide (LPS) and clear unwanted anionic cellular remnants, such as microparticles, from the circulation. Our previous study demonstrated that TLR‑4 and its signaling pathways contribute to the upregulation of pro‑coagulant factors and pro‑inflammatory cytokines in monocytes induced by anti‑β2GPI in vitro. The present study aimed to define the roles of TLR‑4 in vivo. C3H/HeN mice (TLR‑4 intact) and C3H/HeJ mice (TLR‑4 defective) were stimulated with an intraperitoneal injection with anti‑β2GPI‑immunoglobulin G(IgG), then peritoneal macrophages and vascular endothelial cells (VECs) were extracted from treated mice, and analyses were conducted on the expression profiles of pro‑inflammatory cytokines and adhesion molecules. The results demonstrated that the expression of pro‑inflammatory cytokines, including tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑1β and IL‑6, in the peritoneal macrophages, and adhesion molecules, including intercellular cell adhesion molecule‑1 (ICAM‑1), vascular cell adhesion molecule‑1 (VCAM‑1) and E‑selectin, in VECs of C3H/HeN mice (TLR‑4 intact) were significantly higher than those of C3H/HeJ mice (TLR‑4 defective). The phosphorylation levels of p38 mitogen‑activated protein kinase (MAPK) and nuclear factor‑κB (NF‑κB) p65 in peritoneal macrophages and VECs from C3H/HeN mice stimulated with anti‑β2GPI‑IgG were significantly increased compared with those from C3H/HeJ mice (TLR‑4 defective). The isotype control antibody (NR‑IgG) had no such effects on peritoneal macrophages and VECs. Furthermore, the inhibitors of TLR‑4, p38 MAPK and NF‑κB may significantly reduce the anti‑β2GPI‑IgG‑induced TNF‑α, IL‑1β and IL‑6 mRNAs expression in the peritoneal macrophages from TLR‑4 intact mice. The results indicated that a TLR‑4 signal transduction pathway is involved in anti‑β2GPI‑IgG‑induced activation of peritoneal macrophages and VECs. This study has provided a basis for subsequent investigations to elucidate the pathological mechanisms underlying anti‑phospholipid syndrome.

View Figures

View References

1	Chaturvedi S and McCrae KR: Diagnosis and management of the antiphospholipid syndrome. Blood Rev. 31:406–417. 2017. View Article : Google Scholar : PubMed/NCBI
2	McNeil HP, Chesterman CN and Krilis SA: Immunology and clinical importance of antiphospholipid antibodies. Adv Immunol. 49:193–280. 1991. View Article : Google Scholar : PubMed/NCBI
3	Willis R, Harris EN and Pierangeli SS: Pathogenesis of the antiphospholipid syndrome. Semin Thromb Hemost. 38:305–321. 2012. View Article : Google Scholar : PubMed/NCBI
4	Tanne D, Katzav A, Beilin O, Grigoriadis NC, Blank M, Pick CG, Landenberg Pv, Shoenfeld Y and Chapman J: Interaction of inflammation, thrombosis, aspirin and enoxaparin in CNS experimental antiphospholipid syndrome. Neurobiol Dis. 30:56–64. 2008. View Article : Google Scholar : PubMed/NCBI
5	McNeil HP, Simpson RJ, Chesterman CN and Krilis SA: Anti-phospholipid antibodies are directed against a complex antigen that includes a lipid-binding inhibitor of coagulation: Beta 2-glycoprotein I (apolipoprotein H). Proc Natl Acad Sci USA. 87:4120–4124. 1990. View Article : Google Scholar : PubMed/NCBI
6	Galli M, Comfurius P, Maassen C, Hemker HC, de Baets MH, van Breda-Vriesman PJ, Barbui T, Zwaal RF and Bevers EM: Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet. 335:1544–1547. 1990. View Article : Google Scholar : PubMed/NCBI
7	de Laat B, Derksen RH, Urbanus RT and de Groot PG: 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. 105:1540–1545. 2005. View Article : Google Scholar : PubMed/NCBI
8	Forastiero RR, Martinuzzo ME and de Larrañaga GF: Circulating levels of tissue factor and proinflammatory cytokines in patients with primary antiphospholipid syndrome or leprosy related antiphospholipid antibodies. Lupus. 14:129–136. 2005. View Article : Google Scholar : PubMed/NCBI
9	Clemens N, Frauenknecht K, Katzav A, Sommer C and von Landenberg P: In vitro effects of antiphospholipid syndrome-IgG fractions and human monoclonal antiphospholipid IgG antibody on human umbilical vein endothelial cells and monocytes. Ann NY Acad Sci. 1173:805–813. 2009. View Article : Google Scholar : PubMed/NCBI
10	Levi M and van der Poll T: Two-way interactions between inflammation and coagulation. Trends Cardiovasc Med. 15:254–259. 2005. View Article : Google Scholar : PubMed/NCBI
11	ten Cate JW, van der Poll T, Levi M, ten Cate H and van Deventer SJ: Cytokines: Triggers of clinical thrombotic disease. Thromb Haemost. 78:415–419. 1997. View Article : Google Scholar : PubMed/NCBI
12	Meroni PL, Raschi E, Testoni C, Tincani A, Balestrieri G, Molteni R, Khamashta MA, Tremoli E and Camera M: Statins prevent endothelial cell activation induced by antiphospholipid (anti-beta2-glycoprotein I) antibodies: Effect on the proadhesive and proinflammatory phenotype. Arthritis Rheum. 44:2870–2878. 2001. View Article : Google Scholar : PubMed/NCBI
13	Pierangeli SS, Colden-Stanfield M, Liu X, Barker JH, Anderson GL and Harris EN: Antiphospholipid antibodies from antiphospholipid syndrome patients activate endothelial cells in vitro and in vivo. Circulation. 99:1997–2002. 1999. View Article : Google Scholar : PubMed/NCBI
14	Zhou H, Sheng L, Wang H, Xie H, Mu Y, Wang T and Yan J: Anti-β2GPI/β2GPI stimulates activation of THP-1 cells through TLR4/MD-2/MyD88 and NF-κB signaling pathways. Thromb Res. 132:742–749. 2013. View Article : Google Scholar : PubMed/NCBI
15	Vega-Ostertag ME, Ferrara DE, Romay-Penabad Z, Liu X, Taylor WR, Colden-Stanfield M and Pierangeli SS: Role of p38 mitogen-activated protein kinase in antiphospholipid antibody-mediated thrombosis and endothelial cell activation. J Thromb Haemost. 5:1828–1834. 2007. View Article : Google Scholar : PubMed/NCBI
16	Medzhitov R, Preston-Hurlburt P and Janeway CA Jr: A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature. 388:394–397. 1997. View Article : Google Scholar : PubMed/NCBI
17	Medzhitov R and Janeway CA Jr: Innate immunity: The virtues of a nonclonal system of recognition. Cell. 91:295–298. 1997. View Article : Google Scholar : PubMed/NCBI
18	Werling D and Jungi TW: TOLL-like receptors linking innate and adaptive immune response. Vet Immunol Immunopathol. 91:1–12. 2003. View Article : Google Scholar : PubMed/NCBI
19	Xia L, Zhou H, Hu L, Xie H, Wang T, Xu Y, Liu J, Zhang X and Yan J: Both NF-κB and c-Jun/AP-1 involved in anti-β2GPI/β2GPI-induced tissue factor expression in monocytes. Thromb Haemost. 109:643–651. 2013. View Article : Google Scholar : PubMed/NCBI
20	Zhou H, Chen D, Xie H, Xia L, Wang T, Yuan W and Yan J: Activation of MAPKs in the anti-β2GPI/β2GPI-induced tissue factor expression through TLR4/IRAKs pathway in THP-1 cells. Thromb Res. 130:e229–e235. 2012. View Article : Google Scholar : PubMed/NCBI
21	Xie HX, Zhou H, Wang HB, Chen DD, Wang T, Zhang XM, Xia LF and Mu Y: The activation of TRIF-dependent signaling pathway in THP-1 cells induced by β₂ GPI/anti-β₂ GPI antibodies complex. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 27:1280–1283, 1287. 2011.(In Chinese). PubMed/NCBI
22	Ethics and Animal Use, . Guide for the Care and Use of Laboratory Animals. 8th. National Academies Press (US); Washington (DC): 2011, PubMed/NCBI
23	Xie H, Kong X, Zhou H, Xie Y, Sheng L, Wang T, Xia L and Yan J: TLR4 is involved in the pathogenic effects observed in a murine model of antiphospholipid syndrome. Clin Immunol. 160:198–210. 2015. View Article : Google Scholar : PubMed/NCBI
24	He C, Zhang G, Zhou H, Cheng S and Farwa A: Effects of Toll-like receptor 4 on β2-glycoprotein I-induced splenic T cell subsets differentiation. Immunol Lett. 198:17–25. 2018. View Article : Google Scholar : PubMed/NCBI
25	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
26	Brandt KJ, Kruithof EK and de Moerloose P: Receptors involved in cell activation by antiphospholipid antibodies. Thromb Res. 132:408–413. 2013. View Article : Google Scholar : PubMed/NCBI
27	Pierangeli SS, Vega-Ostertag ME, Raschi E, Liu X, Romay-Penabad Z, De Micheli V, Galli M, Moia M, Tincani A, Borghi MO, et al: Toll-like receptor and antiphospholipid mediated thrombosis: In vivo studies. Ann Rheum Dis. 66:1327–1333. 2007. View Article : Google Scholar : PubMed/NCBI
28	Gao MY, Chen L, Yang L, Yu X, Kou JP and Yu BY: Berberine inhibits LPS-induced TF procoagulant activity and expression through NF-κB/p65, Akt and MAPK pathway in THP-1 cells. Pharmacol Rep. 66:480–484. 2014. View Article : Google Scholar : PubMed/NCBI
29	Li X, Zheng Z, Li X and Ma X: Unfractionated heparin inhibits lipopolysaccharide-induced inflammatory response through blocking p38 MAPK and NF-κB activation on endothelial cell. Cytokine. 60:114–121. 2012. View Article : Google Scholar : PubMed/NCBI
30	Willis R and Pierangeli SS: Anti-β2-glycoprotein I antibodies. Ann NY Acad Sci. 1285:44–58. 2013. View Article : Google Scholar : PubMed/NCBI
31	Zhou H, Wolberg AS and Roubey RA: Characterization of monocyte tissue factor activity induced by IgG antiphospholipid antibodies and inhibition by dilazep. Blood. 104:2353–2358. 2004. View Article : Google Scholar : PubMed/NCBI
32	Zhou H, Wang H, Li N, Yu Y, Huang H, Yan Y and Wang T: Annexin A2 mediates anti-beta 2 GPI/beta 2 GPI-induced tissue factor expression on monocytes. Int J Mol Med. 24:557–562. 2009. View Article : Google Scholar : PubMed/NCBI
33	Zhou H, Ling S, Yu Y, Wang T and Hu H: Involvement of Annexin A2 in anti-beta2GPI/beta2GPI-induced tissue factor expression on monocytes. Cell Res. 17:737–739. 2007. View Article : Google Scholar : PubMed/NCBI
34	Zhou H, Yan Y, Xu G, Zhou B, Wen H, Guo D, Zhou F and Wang H: Toll-like receptor (TLR)-4 mediates anti-β2GPI/β2GPI-induced tissue factor expression in THP-1 cells. Clin Exp Immunol. 163:189–198. 2011. View Article : Google Scholar : PubMed/NCBI
35	Xie H, Zhou H, Wang H, Chen D, Xia L, Wang T and Yan J: Anti-β(2)GPI/β(2)GPI induced TF and TNF-α expression in monocytes involving both TLR4/MyD88 and TLR4/TRIF signaling pathways. Mol Immunol. 53:246–254. 2013. View Article : Google Scholar : PubMed/NCBI
36	Hurst J, Lorenz M, Prinz N and von Landenberg P: The roll of Toll-like receptors in the antiphospholipid syndrome. Curr Rheumatol Rep. 12:58–63. 2010. View Article : Google Scholar : PubMed/NCBI
37	Zelaya H, Rothmeier AS and Ruf W: Tissue factor at the crossroad of coagulation and cell signaling. J Thromb Haemost. 16:1941–1952. 2018. View Article : Google Scholar : PubMed/NCBI
38	Pierangeli SS, Espinola RG, Liu X and Harris EN: Thrombogenic effects of antiphospholipid antibodies are mediated by intercellular cell adhesion molecule-1, vascular cell adhesion molecule-1, and P-selectin. Circ Res. 88:245–250. 2001. View Article : Google Scholar : PubMed/NCBI
39	Ghosh TK, Mickelson DJ, Solberg JC, Lipson KE, Inglefield JR and Alkan SS: TLR-TLR cross talk in human PBMC resulting in synergistic and antagonistic regulation of type-1 and 2 interferons, IL-12 and TNF-alpha. Int Immunopharmacol. 7:1111–1121. 2007. View Article : Google Scholar : PubMed/NCBI
40	Benhamou Y, Bellien J, Armengol G, Brakenhielm E, Adriouch S, Iacob M, Remy-Jouet I, Le Cam-Duchez V, Monteil C, Renet S, et al: Role of Toll-like receptors 2 and 4 in mediating endothelial dysfunction and arterial remodeling in primary arterial antiphospholipid syndrome. Arthritis Rheumatol. 66:3210–3220. 2014. View Article : Google Scholar : PubMed/NCBI
41	Yu Y, Zhou H, Xia L, Kong X, Xie Y, Xie H, He C and Cheng S: TLR2 blockade reduces TNF-α expression induced by β2GP1/anti-β2GP1 complex in mouse peritoneal macrophages. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 32:446–450, 456. 2016.(In Chinese). PubMed/NCBI
42	Sikara MP, Routsias JG, Samiotaki M, Panayotou G, Moutsopoulos HM and Vlachoyiannopoulos PG: {beta}2 Glycoprotein I ({beta}2GPI) binds platelet factor 4 (PF4): Implications for the pathogenesis of antiphospholipid syndrome. Blood. 115:713–723. 2010. View Article : Google Scholar : PubMed/NCBI
43	Kawai T and Akira S: Signaling to NF-kappaB by Toll-like receptors. Trends Mol Med. 13:460–469. 2007. View Article : Google Scholar : PubMed/NCBI
44	Olson CM, Hedrick MN, Izadi H, Bates TC, Olivera ER and Anguita J: p38 mitogen-activated protein kinase controls NF-kappaB transcriptional activation and tumor necrosis factor alpha production through RelA phosphorylation mediated by mitogen- and stress-activated protein kinase 1 in response to Borrelia burgdorferi antigens. Infect Immun. 75:270–277. 2007. View Article : Google Scholar : PubMed/NCBI
45	Guo RM, Xu WM, Lin JC, Mo LQ, Hua XX, Chen PX, Wu K, Zheng DD and Feng JQ: Activation of the p38 MAPK/NF-κB pathway contributes to doxorubicin-induced inflammation and cytotoxicity in H9c2 cardiac cells. Mol Med Rep. 8:603–608. 2013. View Article : Google Scholar : PubMed/NCBI
46	Ruiz-Irastorza G and Khamashta MA: Lupus and pregnancy: Integrating clues from the bench and bedside. Eur J Clin Invest. 41:672–678. 2011. View Article : Google Scholar : PubMed/NCBI
47	Matsunaga N, Tsuchimori N, Matsumoto T and Ii M: TAK-242 (resatorvid), a small-molecule inhibitor of Toll-like receptor (TLR) 4 signaling, binds selectively to TLR4 and interferes with interactions between TLR4 and its adaptor molecules. Mol Pharmacol. 79:34–41. 2011. View Article : Google Scholar : PubMed/NCBI
48	Li J, Csakai A, Jin J, Zhang F and Yin H: Therapeutic developments targeting Toll-like receptor-4-mediated neuroinflammation. ChemMedChem. 11:154–165. 2016. View Article : Google Scholar : PubMed/NCBI
49	Comarmond C and Cacoub P: Antiphospholipid syndrome: From pathogenesis to novel immunomodulatory therapies. Autoimmun Rev. 12:752–757. 2013. View Article : Google Scholar : PubMed/NCBI