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J Clin Invest. Dec 1995; 96(6): 2569–2577.
PMCID: PMC185960

Autoantibodies against heat shock protein 60 mediate endothelial cytotoxicity.

Abstract

Stress or heat shock proteins (hsp) are a family of approximately two dozen proteins with a high degree of amino acid sequence homology between different species, ranging from prokaryotes to humans, and are representative of a generalized response to environmental and metabolic stressors. Our previous studies showed increased expression of human hsp60 on endothelial cells of arterial intima with atherosclerotic lesions, and elevated levels of serum antibodies (Ab) against hsp65/60 in subjects with carotid atherosclerosis. To investigate the possible involvement of anti-hsp65/60 Ab in endothelial injury, specific hsp-Ab were isolated from human high titer sera by affinity chromatography and probed on heat-shock human umbilical vein endothelial cells. Purified human anti-hsp65/60 Ab reacted specifically with mycobacterial hsp65, human hsp60, and a 60-kD protein band of heat-shocked endothelial cells. High levels of hsp60 mRNA expression in endothelial cells were found between 4 and 12 h after 30 min treatment at 42 degrees C. In immunofluorescence tests, positive staining of heat-stressed endothelial cells was observed not only in the cytoplasm but also on the cell surface. Furthermore, only heat-stressed, but not untreated, Cr-labeled endothelial cells were lysed by anti-hsp65/60 Ab in the presence of complement (complement-mediated cytotoxicity) or peripheral blood mononuclear cells (antibody-dependent cellular cytotoxicity). Control Abs, including human anti-hsp65/60 low titer antiserum, human Ig fraction deprived of hsp65/60 Ab, and mAbs to Factor VIII, alpha-actin, hsp70, and CD3 showed no cytotoxic effect. In conclusion, human serum anti-hsp65 antibodies act as autoantibodies reacting with hsp60 on stressed endothelial cells and are able to mediate endothelial cytotoxicity. Thus, a humoral immune reaction to hsp60 may play an important role in the pathogenesis of atherosclerosis.

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  • Morimoto RI. Cells in stress: transcriptional activation of heat shock genes. Science. 1993 Mar 5;259(5100):1409–1410. [PubMed]
  • Udelsman R, Blake MJ, Stagg CA, Li DG, Putney DJ, Holbrook NJ. Vascular heat shock protein expression in response to stress. Endocrine and autonomic regulation of this age-dependent response. J Clin Invest. 1993 Feb;91(2):465–473. [PMC free article] [PubMed]
  • Xu Q, Schett G, Seitz CS, Hu Y, Gupta RS, Wick G. Surface staining and cytotoxic activity of heat-shock protein 60 antibody in stressed aortic endothelial cells. Circ Res. 1994 Dec;75(6):1078–1085. [PubMed]
  • Xu Q, Kleindienst R, Waitz W, Dietrich H, Wick G. Increased expression of heat shock protein 65 coincides with a population of infiltrating T lymphocytes in atherosclerotic lesions of rabbits specifically responding to heat shock protein 65. J Clin Invest. 1993 Jun;91(6):2693–2702. [PMC free article] [PubMed]
  • Kleindienst R, Xu Q, Willeit J, Waldenberger FR, Weimann S, Wick G. Immunology of atherosclerosis. Demonstration of heat shock protein 60 expression and T lymphocytes bearing alpha/beta or gamma/delta receptor in human atherosclerotic lesions. Am J Pathol. 1993 Jun;142(6):1927–1937. [PMC free article] [PubMed]
  • Xu Q, Dietrich H, Steiner HJ, Gown AM, Schoel B, Mikuz G, Kaufmann SH, Wick G. Induction of arteriosclerosis in normocholesterolemic rabbits by immunization with heat shock protein 65. Arterioscler Thromb. 1992 Jul;12(7):789–799. [PubMed]
  • Xu Q, Willeit J, Marosi M, Kleindienst R, Oberhollenzer F, Kiechl S, Stulnig T, Luef G, Wick G. Association of serum antibodies to heat-shock protein 65 with carotid atherosclerosis. Lancet. 1993 Jan 30;341(8840):255–259. [PubMed]
  • Xu Q, Luef G, Weimann S, Gupta RS, Wolf H, Wick G. Staining of endothelial cells and macrophages in atherosclerotic lesions with human heat-shock protein-reactive antisera. Arterioscler Thromb. 1993 Dec;13(12):1763–1769. [PubMed]
  • Marder VJ, Mannucci PM, Firkin BG, Hoyer LW, Meyer D. Standard nomenclature for factor VIII and von Willebrand factor: a recommendation by the International Committee on Thrombosis and Haemostasis. Thromb Haemost. 1985 Dec 17;54(4):871–872. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Dugaiczyk A, Haron JA, Stone EM, Dennison OE, Rothblum KN, Schwartz RJ. Cloning and sequencing of a deoxyribonucleic acid copy of glyceraldehyde-3-phosphate dehydrogenase messenger ribonucleic acid isolated from chicken muscle. Biochemistry. 1983 Mar 29;22(7):1605–1613. [PubMed]
  • Singh B, Gupta RS. Expression of human 60-kD heat shock protein (HSP60 or P1) in Escherichia coli and the development and characterization of corresponding monoclonal antibodies. DNA Cell Biol. 1992 Jul-Aug;11(6):489–496. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Sgonc R, Boeck G, Dietrich H, Gruber J, Recheis H, Wick G. Simultaneous determination of cell surface antigens and apoptosis. Trends Genet. 1994 Feb;10(2):41–42. [PubMed]
  • Jürgens G, Xu QB, Huber LA, Böck G, Howanietz H, Wick G, Traill KN. Promotion of lymphocyte growth by high density lipoproteins (HDL). Physiological significance of the HDL binding site. J Biol Chem. 1989 May 25;264(15):8549–8556. [PubMed]
  • Salonen JT, Ylä-Herttuala S, Yamamoto R, Butler S, Korpela H, Salonen R, Nyyssönen K, Palinski W, Witztum JL. Autoantibody against oxidised LDL and progression of carotid atherosclerosis. Lancet. 1992 Apr 11;339(8798):883–887. [PubMed]
  • Seifert PS, Kazatchkine MD. The complement system in atherosclerosis. Atherosclerosis. 1988 Oct;73(2-3):91–104. [PubMed]
  • Libby P, Hansson GK. Involvement of the immune system in human atherogenesis: current knowledge and unanswered questions. Lab Invest. 1991 Jan;64(1):5–15. [PubMed]
  • Muscari A, Bozzoli C, Gerratana C, Zaca' F, Rovinetti C, Zauli D, La Placa M, Puddu P. Association of serum IgA and C4 with severe atherosclerosis. Atherosclerosis. 1988 Nov;74(1-2):179–186. [PubMed]
  • Hollander W, Colombo MA, Kirkpatrick B, Paddock J. Soluble proteins in the human atherosclerotic plaque. With spectral reference to immunoglobulins, C3-complement component, alpha 1-antitrypsin and alpha 2-macroglobulin. Atherosclerosis. 1979 Dec;34(4):391–405. [PubMed]
  • Vlaicu R, Niculescu F, Rus HG, Cristea A. Immunohistochemical localization of the terminal C5b-9 complement complex in human aortic fibrous plaque. Atherosclerosis. 1985 Nov;57(2-3):163–177. [PubMed]
  • Seifert PS, Hansson GK. Complement receptors and regulatory proteins in human atherosclerotic lesions. Arteriosclerosis. 1989 Nov-Dec;9(6):802–811. [PubMed]
  • Seifert PS, Hugo F, Hansson GK, Bhakdi S. Prelesional complement activation in experimental atherosclerosis. Terminal C5b-9 complement deposition coincides with cholesterol accumulation in the aortic intima of hypercholesterolemic rabbits. Lab Invest. 1989 Jun;60(6):747–754. [PubMed]
  • Seifert PS, Hugo F, Tranum-Jensen J, Zâhringer U, Muhly M, Bhakdi S. Isolation and characterization of a complement-activating lipid extracted from human atherosclerotic lesions. J Exp Med. 1990 Aug 1;172(2):547–557. [PMC free article] [PubMed]
  • Ylä-Herttuala S, Palinski W, Butler SW, Picard S, Steinberg D, Witztum JL. Rabbit and human atherosclerotic lesions contain IgG that recognizes epitopes of oxidized LDL. Arterioscler Thromb. 1994 Jan;14(1):32–40. [PubMed]
  • Jürgens G, Chen Q, Esterbauer H, Mair S, Ledinski G, Dinges HP. Immunostaining of human autopsy aortas with antibodies to modified apolipoprotein B and apoprotein(a). Arterioscler Thromb. 1993 Nov;13(11):1689–1699. [PubMed]
  • Stemme S, Faber B, Holm J, Wiklund O, Witztum JL, Hansson GK. T lymphocytes from human atherosclerotic plaques recognize oxidized low density lipoprotein. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3893–3897. [PMC free article] [PubMed]
  • Jindal S, Burke E, Bettencourt B, Khandekar S, Young RA, Dwyer DS. Human Hsp60: bacterial expression, purification, development of monoclonal antibodies and a sandwich ELISA for quantitation. Immunol Lett. 1994 Feb;39(2):127–135. [PubMed]
  • Evans DJ, Norton P, Ivanyi J. Distribution in tissue sections of the human groEL stress-protein homologue. APMIS. 1990 May;98(5):437–441. [PubMed]
  • Davies PF, Remuzzi A, Gordon EJ, Dewey CF, Jr, Gimbrone MA., Jr Turbulent fluid shear stress induces vascular endothelial cell turnover in vitro. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2114–2117. [PMC free article] [PubMed]
  • Kiessling R, Grönberg A, Ivanyi J, Söderström K, Ferm M, Kleinau S, Nilsson E, Klareskog L. Role of hsp60 during autoimmune and bacterial inflammation. Immunol Rev. 1991 Jun;121:91–111. [PubMed]
  • Wand-Württenberger A, Schoel B, Ivanyi J, Kaufmann SH. Surface expression by mononuclear phagocytes of an epitope shared with mycobacterial heat shock protein 60. Eur J Immunol. 1991 Apr;21(4):1089–1092. [PubMed]
  • Di Cesare S, Poccia F, Mastino A, Colizzi V. Surface expressed heat-shock proteins by stressed or human immunodeficiency virus (HIV)-infected lymphoid cells represent the target for antibody-dependent cellular cytotoxicity. Immunology. 1992 Jun;76(2):341–343. [PMC free article] [PubMed]
  • Palinski W, Miller E, Witztum JL. Immunization of low density lipoprotein (LDL) receptor-deficient rabbits with homologous malondialdehyde-modified LDL reduces atherogenesis. Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):821–825. [PMC free article] [PubMed]
  • Shimano H, Yamada N, Ishibashi S, Mokuno H, Mori N, Gotoda T, Harada K, Akanuma Y, Murase T, Yazaki Y, et al. Oxidation-labile subfraction of human plasma low density lipoprotein isolated by ion-exchange chromatography. J Lipid Res. 1991 May;32(5):763–773. [PubMed]
  • Hodis HN, Kramsch DM, Avogaro P, Bittolo-Bon G, Cazzolato G, Hwang J, Peterson H, Sevanian A. Biochemical and cytotoxic characteristics of an in vivo circulating oxidized low density lipoprotein (LDL-). J Lipid Res. 1994 Apr;35(4):669–677. [PubMed]
  • Cohen IR. A heat shock protein, molecular mimicry and autoimmunity. Isr J Med Sci. 1990 Dec;26(12):673–676. [PubMed]
  • Zinkernagel RM, Cooper S, Chambers J, Lazzarini RA, Hengartner H, Arnheiter H. Virus-induced autoantibody response to a transgenic viral antigen. Nature. 1990 May 3;345(6270):68–71. [PubMed]
  • Schattner A, Rager-Zisman B. Virus-induced autoimmunity. Rev Infect Dis. 1990 Mar-Apr;12(2):204–222. [PubMed]
  • Coulie PG, Van Snick J. Rheumatoid factor (RF) production during anamnestic immune responses in the mouse. III. Activation of RF precursor cells is induced by their interaction with immune complexes and carrier-specific helper T cells. J Exp Med. 1985 Jan 1;161(1):88–97. [PMC free article] [PubMed]
  • Emeson EE, Robertson AL., Jr T lymphocytes in aortic and coronary intimas. Their potential role in atherogenesis. Am J Pathol. 1988 Feb;130(2):369–376. [PMC free article] [PubMed]
  • Hansson GK, Jonasson L, Lojsthed B, Stemme S, Kocher O, Gabbiani G. Localization of T lymphocytes and macrophages in fibrous and complicated human atherosclerotic plaques. Atherosclerosis. 1988 Aug;72(2-3):135–141. [PubMed]
  • Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993 Apr 29;362(6423):801–809. [PubMed]
  • Xu QB, Oberhuber G, Gruschwitz M, Wick G. Immunology of atherosclerosis: cellular composition and major histocompatibility complex class II antigen expression in aortic intima, fatty streaks, and atherosclerotic plaques in young and aged human specimens. Clin Immunol Immunopathol. 1990 Sep;56(3):344–359. [PubMed]
  • Hansson GK, Holm J, Holm S, Fotev Z, Hedrich HJ, Fingerle J. T lymphocytes inhibit the vascular response to injury. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10530–10534. [PMC free article] [PubMed]
  • Fyfe AI, Qiao JH, Lusis AJ. Immune-deficient mice develop typical atherosclerotic fatty streaks when fed an atherogenic diet. J Clin Invest. 1994 Dec;94(6):2516–2520. [PMC free article] [PubMed]
  • Jones DB, Coulson AF, Duff GW. Sequence homologies between hsp60 and autoantigens. Immunol Today. 1993 Mar;14(3):115–118. [PubMed]
  • Wick G, Schett G, Amberger A, Kleindienst R, Xu Q. Is atherosclerosis an immunologically mediated disease? Immunol Today. 1995 Jan;16(1):27–33. [PubMed]

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