Advertisement

Cell Stress and Chaperones

, Volume 18, Issue 4, pp 483–493 | Cite as

Evidence of a role for both anti-Hsp70 antibody and endothelial surface membrane Hsp70 in atherosclerosis

  • Xue Leng
  • Xinxing Wang
  • Wei Pang
  • Rui Zhan
  • Zhiqing Zhang
  • Liqun Wang
  • Xiujie Gao
  • Lingjia QianEmail author
Original Paper

Abstract

Although previous studies have shown that autoantigens such as Hsps have been implicated by induction of an autoimmune process in the development of atherosclerosis, the exact role of anti-Hsp70 antibody in atherosclerosis is unknown. In the present study, the levels of anti-Hsp70 autoantibodies and oxidized low density lipoprotein (OxLDL) were all significantly increased, and they were strongly correlated in an atherosclerosis model. After the endothelial cells were incubated with 20 μg/mL OxLDL for 12 h at 37 °C and followed by 90 min recovery, Hsp70 positive staining of OxLDL-treated endothelial cells was observed on the cell surface in immunostaining and flow cytometric analysis. This membrane Hsp70 was not from culture supernatant Hsp70 and binding of extracellular Hsp70 but was defined as endothelial surface membrane Hsp70. Furthermore, only in the OxLDL-treated group, but not in the untreated group, 51Cr-labeled endothelial cells were lysed by anti-Hsp70 antibody (BD091, IgAS) in the presence of either complement or peripheral blood mononuclear cells. Control antibodies, including IgNor, mAb to Hsp70 (SPA-810), and mAbs to Factor VIII, α-actin, and CD3 showed no cytotoxic effects. In conclusion, anti-Hsp70 antibodies could be reacting with the endothelial surface membrane Hsp70 induced by OxLDL and were able to mediate endothelial cytotoxicity. There is a possibility that a humoral immune reaction to endothelial surface membrane Hsp70 may play an important role in the pathogenesis of atherosclerosis.

Keywords

Endothelial cell Surface membrane Hsp70 Anti-Hsp70 antibody OxLDL Atherosclerosis 

Abbreviations

Hsps

Heat shock proteins

OxLDL

Oxidized low density lipoprotein

AS

Atherosclerosis

IgAS

Atherosclerotic rat plasma Hsp70 autoantibodies

IgNor

Control rat plasma Hsp70 autoantibodies

mAb

Monoclonal antibody

BD091

Monoclonal antibody to Hsp70 (which recognized Hsp70 C-terminal)

SPA-810

Monoclonal antibody to Hsp70 (binding site, aa437-504)

PBS

Phosphate-buffered saline

HRP

Horseradish peroxidase

TMB

Tetramethyl benzidine

ELISA

Enzyme-linked immunosorbent assay

Notes

Acknowledgments

This research was supported by Important National Science & Technology Specific Projects (grant no.2009ZX09103-629) and by the National Natural Science Foundation of China (grant nos. 30800456 and 81000584).

Supplementary material

12192_2013_404_MOESM1_ESM.ppt (153 kb)
ESM 1 (PPT 153 kb)

References

  1. Arispe N, Doh M, De Maio A (2002) Lipid interaction differentiates the constitutive and stress-induced heat shock proteins Hsc70 and Hsp70. Cell Stress Chaperones 7:330–338PubMedCrossRefGoogle Scholar
  2. Arispe N, Doh M, Simakova O, Kurganov B, De Maio A (2004) Hsc70 and Hsp70 interact with phosphatidylserine on the surface of PC12 cells resulting in a decrease of viability. FASEB J 18:1636–1645PubMedCrossRefGoogle Scholar
  3. Bauer HC, Hurtenbach U (1986) Murine cortical brain cells are autoantigenic from a distinct developmental stage onwards. J Neuroimmunol 12:1–13PubMedCrossRefGoogle Scholar
  4. Cai GJ, Miao CY, Xie HH, Lu LH, Su DF (2005) Arterial baroreflex dysfunction promotes atherosclerosis in rats. Atherosclerosis 183:41–47PubMedCrossRefGoogle Scholar
  5. Campisi J, Leem TH, Fleshner M (2003) Stress-induced extracellular Hsp72 is a functionally significant danger signal to the immune system. Cell Stress Chaperones 8:272–286PubMedCrossRefGoogle Scholar
  6. Chan YCSN, Abdus-Samee M et al (1999) Anti-heat-shock protein 70 kDa antibodies in vascular patients. Eur J Vasc Endovasc Surg 18:381–385PubMedCrossRefGoogle Scholar
  7. Dulin E, Garcia-Barreno P, Guisasola MC (2010) Extracellular heat shock protein 70 (HSPA1A) and classical vascular risk factors in a general population. Cell Stress Chaperones 15:929–937PubMedCrossRefGoogle Scholar
  8. Enomoto Y, Bharti A, Khaleque AA, Song B, Liu C, Apostolopoulos V, Xing PX, Calderwood SK, Gong J (2006) Enhanced immunogenicity of heat shock protein 70 peptide complexes from dendritic cell-tumor fusion cells. J Immunol 177:5946–5955PubMedGoogle Scholar
  9. Gehrmann M, Liebisch G, Schmitz G, Anderson R, Steinem C, De Maio A, Pockley G, Multhoff G (2008) Tumor-specific Hsp70 plasma membrane localization is enabled by the glycosphingolipid Gb3. PLoS One 3:e1925PubMedCrossRefGoogle Scholar
  10. George J, Greenberg S, Barshack I, Singh M, Pri-Chen S, Laniado S, Keren G (2001) Accelerated intimal thickening in carotid arteries of balloon-injured rats after immunization against heat shock protein 70. J Am Coll Cardiol 38:1564–1569PubMedCrossRefGoogle Scholar
  11. Guzhova I, Kislyakova K, Moskaliova O, Fridlanskaya I, Tytell M, Cheetham M, Margulis B (2001) In vitro studies show that Hsp70 can be released by glia and that exogenous Hsp70 can enhance neuronal stress tolerance. Brain Res 914:66–73PubMedCrossRefGoogle Scholar
  12. Hightower LE, Guidon PT Jr (1989) Selective release from cultured mammalian cells of heat-shock (stress) proteins that resemble glia-axon transfer proteins. J Cell Physiol 138:257–266PubMedCrossRefGoogle Scholar
  13. Huber LA, Scheffler E, Poll T, Ziegler R, Dresel HA (1990) 17 beta-estradiol inhibits LDL oxidation and cholesteryl ester formation in cultured macrophages. Free Radic Res Commun 8:167–173PubMedCrossRefGoogle Scholar
  14. Jialal I, Devaraj S (1996) The role of oxidized low density lipoprotein in atherogenesis. J Nutr 126:1053S–1057SPubMedGoogle Scholar
  15. Jurgens G, Xu QB, Huber LA, Bock G, Howanietz H, Wick G, Traill KN (1989) Promotion of lymphocyte growth by high density lipoproteins (HDL). Physiological significance of the HDL binding site. J Biol Chem 264:8549–8556PubMedGoogle Scholar
  16. Kocsis J, Veres A, Vatay A, Duba J, Karadi I, Fust G, Prohaszka Z (2002) Antibodies against the human heat shock protein hsp70 in patients with severe coronary artery disease. Immunol Investig 31:219–231CrossRefGoogle Scholar
  17. Kreisel D, Krupnick AS, Szeto WY, Popma SH, Sankaran D, Krasinskas AM, Amin KM, Rosengard BR (2001) A simple method for culturing mouse vascular endothelium. J Immunol Methods 254:31–45PubMedCrossRefGoogle Scholar
  18. Leng X, Zhan R, Wang Y, Liu X, Gong J, Gao X, Wu L, Wang L, Zhao Y, Wang X, Zhang Z, Pang W, Qian LJ (2010) Anti-heat shock protein 70 autoantibody epitope changes and BD091 promotes atherosclerosis in rats. Cell Stress Chaperones 15:947–958PubMedCrossRefGoogle Scholar
  19. Mayr M, Metzler B, Kiechl S, Willeit J, Schett G, Xu Q, Wick G (1999) Endothelial cytotoxicity mediated by serum antibodies to heat shock proteins of Escherichia coli and Chlamydia pneumoniae: immune reactions to heat shock proteins as a possible link between infection and atherosclerosis. Circulation 99:1560–1566PubMedCrossRefGoogle Scholar
  20. Multhoff G, Hightower LE (1996) Cell surface expression of heat shock proteins and the immune response. Cell Stress Chaperones 1:167–176PubMedCrossRefGoogle Scholar
  21. Muscari A, Bozzoli C, Gerratana C, Zaca F, Rovinetti C, Zauli D, La Placa M, Puddu P (1988) Association of serum IgA and C4 with severe atherosclerosis. Atherosclerosis 74:179–186PubMedCrossRefGoogle Scholar
  22. Pockley A, De U, Kiessling R, Lemne C, Thulin T, Frostegård J (2002) Circulating heat shock protein and heat shock protein antibody levels in established hypertension. J Hypertens 20(9):1815–20PubMedCrossRefGoogle Scholar
  23. Pockley AG, Georgiades A, Thulin T et al (2003) Serum heat shock protein 70 levels predict the development of atherosclerosis in subjects with established hypertension. Hypertension 42:235–238PubMedCrossRefGoogle Scholar
  24. Prohaszka Z, Duba J, Horvath L, Csaszar A, Karadi I, Szebeni A, Singh M, Fekete B, Romics L, Fust G (2001) Comparative study on antibodies to human and bacterial 60 kDa heat shock proteins in a large cohort of patients with coronary heart disease and healthy subjects. Eur J Clin Investig 31:285–292CrossRefGoogle Scholar
  25. Salonen JT, Yla-Herttuala S, Yamamoto R, Butler S, Korpela H, Salonen R, Nyyssonen K, Palinski W, Witztum JL (1992) Autoantibody against oxidised LDL and progression of carotid atherosclerosis. Lancet 339:883–887PubMedCrossRefGoogle Scholar
  26. Sanson M, Auge N, Vindis C, Muller C, Bando Y, Thiers JC, Marachet MA, Zarkovic K, Sawa Y, Salvayre R, Negre-Salvayre A (2009) Oxidized low-density lipoproteins trigger endoplasmic reticulum stress in vascular cells: prevention by oxygen-regulated protein 150 expression. Circ Res 104:328–336PubMedCrossRefGoogle Scholar
  27. Schett G, Xu Q, Amberger A, Van der Zee R, Recheis H, Willeit J, Wick G (1995) Autoantibodies against heat shock protein 60 mediate endothelial cytotoxicity. J Clin Invest 96(6):2569–2577PubMedCrossRefGoogle Scholar
  28. Singh-Jasuja H, Hilf N, Arnold-Schild D, Schild H (2001) The role of heat shock proteins and their receptors in the activation of the immune system. Biol Chem 382:629–636PubMedCrossRefGoogle Scholar
  29. Srivastava P (2002) Roles of heat-shock proteins in innate and adaptive immunity. Nat Rev Immunol 2:185–194PubMedCrossRefGoogle Scholar
  30. Triantafilou M, Miyake K, Golenbock DT, Triantafilou K (2002) Mediators of innate immune recognition of bacteria concentrate in lipid rafts and facilitate lipopolysaccharide-induced cell activation. J Cell Sci 115:2603–2611PubMedGoogle Scholar
  31. Vasankari T, Ahotupa M, Toikka J, Mikkola J, Irjala K, Pasanen P, Neuvonen K, Raitakari O, Viikari J (2001) Oxidized LDL and thickness of carotid intima-media are associated with coronary atherosclerosis in middle-aged men: lower levels of oxidized LDL with statin therapy. Atherosclerosis 155:403–412PubMedCrossRefGoogle Scholar
  32. Wick G, Knoflach M, Xu Q (2004) Autoimmune and inflammatory mechanisms in atherosclerosis. Annu Rev Immunol 22:361–403PubMedCrossRefGoogle Scholar
  33. Xu Q, Schett G, Seitz CS, Hu Y, Gupta RS, Wick G (1994) Surface staining and cytotoxic activity of heat-shock protein 60 antibody in stressed aortic endothelial cells. Circ Res 75:1078–1085PubMedCrossRefGoogle Scholar
  34. Xu Q, Kiechl S, Mayr M, Metzler B, Egger G, Oberhollenzer F, Willeit J, Wick G (1999) Association of serum antibodies to heat-shock protein 65 with carotid atherosclerosis: clinical significance determined in a follow-up study. Circulation 100:1169–1174PubMedCrossRefGoogle Scholar
  35. Xu Q (2002) Role of heat shock proteins in atherosclerosis. Arterioscler Thromb Vasc Biol 22:1547–1559PubMedCrossRefGoogle Scholar
  36. Yuan J, Yang M, Yao H, Zheng J, Yang Q, Chen S, Wei Q, Tanguay RM, Wu T (2005) Plasma antibodies to heat shock protein 60 and heat shock protein 70 are associated with increased risk of electrocardiograph abnormalities in automobile workers exposed to noise. Cell Stress Chaperones 10:126–135PubMedCrossRefGoogle Scholar
  37. Zhang X, Tanguay RM, He M, Deng Q, Miao X, Zhou L, Wu T (2011) Variants of HSPA1A in combination with plasma Hsp70 and anti-Hsp70 antibody levels associated with higher risk of acute coronary syndrome. Cardiology 119:57–64PubMedCrossRefGoogle Scholar
  38. Zhu J, Quyyumi AA, Wu H, Csako G, Rott D, Zalles-Ganley A, Ogunmakinwa J, Halcox J, Epstein SE (2003) Increased serum levels of heat shock protein 70 are associated with low risk of coronary artery disease. Arterioscler Thromb Vasc Biol 23:1055–1059PubMedCrossRefGoogle Scholar
  39. Zhu J, Katz RJ, Quyyumi AA, Canos DA, Rott D, Csako G, Zalles-Ganley A, Ogunmakinwa J, Wasserman AG, Epstein SE (2004) Association of serum antibodies to heat-shock protein 65 with coronary calcification levels: suggestion of pathogen-triggered autoimmunity in early atherosclerosis. Circulation 109:36–41PubMedCrossRefGoogle Scholar
  40. Zhu W, Roma E, Pellegatta F, Catapano AL (1994) Biochem Biophys Res Commun 200:389–394PubMedCrossRefGoogle Scholar

Copyright information

© Cell Stress Society International 2013

Authors and Affiliations

  • Xue Leng
    • 2
  • Xinxing Wang
    • 1
  • Wei Pang
    • 3
  • Rui Zhan
    • 1
  • Zhiqing Zhang
    • 3
  • Liqun Wang
    • 3
  • Xiujie Gao
    • 3
  • Lingjia Qian
    • 1
    Email author
  1. 1.Institute of Basic Medical SciencesBeijingPeople’s Republic of China
  2. 2.Institute of Medical EquipmentTianjinPeople’s Republic of China
  3. 3.Institute of Health & Environmental MedicineTianjinPeople’s Republic of China

Personalised recommendations