Impairment of vascular function following BCG immunisation is associated with immune responses to HSP-60 in the cholesterol-fed rabbit


      An immune response to heat shock protein (HSP)-60/65 has recently been implicated in atherogenesis. The aim of this study was to determine whether this effect may be mediated by impairment of endothelial function. Rabbits were injected with bacillus Calmette-Guerin (BCG) vaccine (n=12) or saline (n=12). A further injection of BCG or saline was administered after 2 weeks. After a further 2 weeks, animals were fed either a 0.25–1% cholesterol diet or a chow diet for 16 weeks. Blood cholesterol levels were maintained at 10–12 mmol/l by altering the dietary cholesterol content. Plasma levels of anti-mycobacterial antibodies rose following BCG immunisation, but anti-HSP antibodies developed only in the BCG-immunised, cholesterol-fed rabbits. Aortic endothelium from cholesterol-fed, but not chow-fed, rabbits stained positively for HSP-60, independently of the immunisation protocol. Endothelial function was impaired in the BCG immunised, cholesterol-fed rabbits as measured by acetylcholine-mediated relaxation of isolated non-atherosclerotic carotid artery rings (P<0.05). This impairment was positively associated with the level of plasma anti-HSP-60 antibodies (P<0.01). These results suggest that BCG immunisation impairs endothelial responses, at least in part, by immune responses against mycobacterial and vascular HSP.


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        • Lamb D.J.
        • El-Sankary W.
        • Ferns G.A.A.
        Molecular mimicry in atherosclerosis: a role for heat shock proteins in immunisation.
        Atherosclerosis. 2002; 167: 177-185
        • Xu Q.
        Role of heat shock proteins in atherosclerosis.
        Arterioscler. Thromb. Vasc. Biol. 2002; 22: 1547-1559
        • Epstein S.E.
        • Zhu J.
        • Burnett M.S.
        • Zhou Y.F.
        • Vercellotti G.
        • Hajjar D.
        Infection and atherosclerosis: potential roles of pathogen burden and molecular mimicry.
        Arterioscler. Thromb. Vasc. Biol. 2000; 20: 1417-1420
        • Schett G.
        • Xu Q.
        • Amberger A.
        • van der Zee R.
        • Recheis H.
        • Willeit J.
        • et al.
        Autoantibodies against heat shock protein 60 mediate endothelial cytotoxicity.
        J. Clin. Invest. 1995; 96: 2569-2577
        • Metzler B.
        • Schett G.
        • Kleindienst R.
        • van der Zee R.
        • Ottenhoff T.
        • Hajeer A.
        • et al.
        Epitope specificity of anti-heat shock protein 60/65 serum antibodies in atherosclerosis.
        Arterioscler. Thromb. Vasc. Biol. 1997; 17: 536-541
        • Henderson A.H.
        Endothelium in control.
        Br. Heart J. 1991; 65: 116-125
        • Boger R.H.
        • Bode-Boger S.M.
        • Frolich J.C.
        The l-arginine-nitric oxide pathway: role in atherosclerosis and therapeutic implications.
        Atherosclerosis. 1996; 127: 1-11
        • Ludmer P.L.
        • Selwyn A.P.
        • Shook T.L.
        • Wayne R.R.
        • Mudge G.H.
        • Alexander R.W.
        • et al.
        Paradoxical vasoconstriction induced by acetylcholine in atherosclerotic coronary arteries.
        N. Eng. J. Med. 1986; 315: 1046-1051
        • Vita J.A.
        • Treasure C.B.
        • Nabel E.G.
        • McLenachon J.M.
        • Fish R.D.
        • Yeung A.C.
        • et al.
        Coronary vasomotor response to acetylcholine relates to risk factors for coronary artery disease.
        Circulation. 1990; 81: 491-497
        • Celermajer D.S.
        • Sorensen K.E.
        • Georgakopoulos D.
        • Bull C.
        • Thomas O.
        • Robinson J.
        • et al.
        Cigarette smoking is associated with dose-related and potentially reversible impairment of endothelium-dependent dilation in healthy young adults.
        Circulation. 1993; 88: 2149-2155
        • Panza J.A.
        • Quyyumi A.A.
        • Callahan T.S.
        • Epstein S.E.
        Effect of antihypertensive treatment on endothelium-dependent vascular relaxation in patients with essential hypertension.
        J. Am. Coll. Cardiol. 1993; 21: 1145-1151
        • Ladel C.H.
        • Hess J.
        • Daugelat S.
        • Mombaerts P.
        • Toneegawa S.
        • Kaufmann S.H.E.
        Contribution of α/β and γ/δ T-lymphocytes to immunity against Mycobacterium bovis bacillus Calmette-Guerin: studies with T-cell receptor-deficient mice.
        Eur. J. Immunol. 1995; 25: 838-846
        • Jackson A.M.
        • Lien E.
        • Alexandroff A.B.
        • Prescott S.
        • Espenik T.
        • James K.
        • et al.
        Soluble urinary CD14 after intravesical bacille Calmette-Guerin immunotherapy for carcinoma in situ.
        Br. J. Urol. 1997; 80: 766-771
        • Hanano R.
        • Kaufmann S.H.
        Nitric oxide production and mycobacterial growth inhibition by murine alveolar macrophages: the sequence of rIFN-gamma stimulation and Mycobacterium bovis BCG infection determines macrophage activation.
        Immunol. Lett. 1995; 45: 23-27
        • Tokunaga T.
        • Yano O.
        • Kuramoto E.
        • Kimura Y.
        • Yamamoto T.
        • Kataoka T.
        • et al.
        Synthetic oligonucleotides with particular base sequences from the cDNA encoding proteins of Mycobacterium bovis BCG induce interferons and activate natural killer cells.
        Microbiol. Immunol. 1992; 36: 55-66
        • Sander B.
        • Skansen-Saphir U.
        • Damm O.
        • Hakansson L.
        • Andersson U.
        Sequential production of Th1 and Th2 cytokines in response to live bacillus Calmette-Guerin.
        Immunology. 1995; 86: 512-518
        • Mustafa A.S.
        • Lundin K.E.
        • Oftung F.
        Human T cells recognize mycobacterial heat shock proteins in the context of multiple HLA-DR molecules: studies with healthy subjects vaccinated with Mycobacterium bovis BCG and Mycobacterium leprae.
        Infect. Immun. 1993; 61: 5294-5301
        • Lamb D.J.
        • Eales L.J.
        • Ferns G.A.A.
        Immunisation with bacillus Calmette-Guerin vaccine increases aortic atherosclerosis in the cholesterol-fed rabbit.
        Atherosclerosis. 1999; 143: 105-113
        • Lamb D.J.
        • Ferns G.A.A.
        The magnitude of the immune response to heat shock protein-65 following BCG immunisation is associated with the extent of experimental atherosclerosis.
        Atherosclerosis. 2002; 165: 231-240
        • Liao F.
        • Andalibi A.
        • de Beer F.C.
        • Fogelman A.M.
        • Lusis A.J.
        Genetic control of inflammatory gene induction and NF-κB-like transcription factor activation in response to an atherogenic diet in mice.
        J. Clin. Invest. 1993; 91: 2572-2579
        • Paris D.
        • Town T.
        • Humphrey J.
        • Yokota K.
        • Mullan M.
        Cholesterol modulates vascular reactivity to endothelin-1 by stimulating a pro-inflammatory pathway.
        Biochem. Biophys. Res. Commun. 2000; 274: 553-558
        • Hughes D.A.
        • Townsend P.J.
        • Haslam P.L.
        Enhancement of the antigen-presenting function of monocytes by cholesterol: possible relevance to inflammatory mechanisms in extrinsic allergic alveolitis and atherosclerosis.
        Clin. Exp. Immunol. 1992; 87: 279-286
        • Alderman C.J.
        • Bunyard P.R.
        • Chain B.M.
        • Foreman J.C.
        • Leake D.S.
        • Katz D.R.
        Effects of oxidised low density lipoprotein on dendritic cells: a possible immunoregulatory component of the atherogenic micro-environment.
        Cardiovasc. Res. 2002; 55: 806-819
        • Khan I.U.
        • Wallin R.
        • Gupta R.S.
        • Kammer G.M.
        Protein kinase A-catalyzed phosphorylation of heat shock protein 60 chaperone regulates its attachment to histone 2B in the T lymphocyte plasma membrane.
        Proc. Natl. Acad. Sci. U.S.A. 1998; 95: 10425-10430
        • Kol A.
        • Bourcier T.
        • Lichtman A.H.
        • Libby P.
        Chlamydial and human heat shock protein 60s activate human vascular endothelium, smooth muscle cells, and macrophages.
        J. Clin. Invest. 1999; 103: 571-577
        • Schett G.
        • Metzler B.
        • Kleindienst R.
        • Amberger A.
        • Recheis H.
        • Xu Q.
        • et al.
        Myocardial injury leads to a release of heat shock protein (HSP)-60 and a suppression of the anti-hsp65 immune response.
        Cardiovasc. Res. 1999; 42: 685-695
        • Kleindienst R.
        • Xu Q.
        • Willeit J.
        • Waldenberger F.R.
        • Weimann S.
        • Wick G.
        Immunology of atherosclerosis: demonstration of heat shock protein-60 expression and T lymphocytes bearing α/β or γ/δ receptor in human atherosclerotic lesions.
        Am. J. Pathol. 1993; 142: 1927-1937
        • Matz J.
        • Andersson T.L.
        • Ferns G.A.A.
        • Anggard E.E.
        Dietary vitamin E increases the resistance to lipoprotein oxidation and attenuates endothelial dysfunction in the cholesterol-fed rabbit.
        Atherosclerosis. 1994; 110: 241-249
        • Xu Q.
        • Dietrich H.
        • Steiner H.J.
        • Gown A.M.
        • Schoel B.
        • Mikuz G.
        • et al.
        Induction of atherosclerosis in normocholesterolaemic rabbits by immunisation with heat shock protein 65.
        Arterioscler. Thromb. Vasc. Biol. 1992; 12: 789-799