Changes of serum antibodies to heat-shock protein 65 in coronary heart disease and acute myocardial infarction

doi:10.1016/0021-9150(96)05931-X

Atherosclerosis

Volume 126, Issue 2, 25 October 1996, Pages 333-338

https://doi.org/10.1016/0021-9150(96)05931-X Get rights and content

Abstract

Accumulating evidence indicates the involvement of heat shock proteins (hsp), a family of stress-inducible proteins, in atherosclerosis. For carotid atherosclerosis an association with an increase in hsp65 antibodies has been demonstrated. To investigate whether such antibodies are also associated with coronary heart disease (CHD) and acute myocardial infarction (MI), an age- and sex-matched study with patients suffering from CHD (n = 114) and MI (n = 89) and healthy controls (n = 76) was performed. All study participants (n = 279) were consecutively recruited according to typical diagnostic criteria. Determination of antibody titres to hsp65 was performed by an enzyme-linked immunosorbent assay (ELISA). Hsp65 antibody titres in CHD showed a significant increase compared to the healthy control group (P = 0.029), however, hsp65 antibody titres were found to be significantly lower in acute MI, compared to CHD (P = 0.005). Alteration in hsp65 antibody titres showed no correlation to established cardiovascular risk factors, e.g. serum total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides, blood pressure, smoking, alcohol intake and body weight. In conclusion, serum concentrations of hsp65 antibodies were elevated independently in coronary heart diseases and declined in patients with acute myocardial infarction, indicating a possible involvement of the antibodies in the pathogenesis of this disease.

Introduction

Atherosclerosis is a disease with a multifactorial origin. Beside the well-established risk factors, smoking, dyslipidemia, hypertension and diabetes mellitus, immune and autoimmune mechanisms have been shown to influence the atherogenic process [1]. The involvement of heat shock proteins (hsp), a family of stress-inducible proteins, could be observed in the development of atheromatous lesions 2, 3, 4. Previous reports described that human hsp65 antibodies highly crossreact to mammalian hsp60 and specifically recognize a 60 kDa protein from homogenates of atherosclerotic lesions [5]. In atherosclerotic lesions, increased expression of hsp60 was observed in endothelial cells and macrophages [6]. Hsp expression in arterial cells can be induced by arterial injury [7], cytokines [8]and oxidized-LDL 9, 10. With respect to the clinical relevance of hsp, it has been demonstrated that serum antibody titres to hsp65 are increased in carotid atherosclerosis [3].

With respect to the development of atherosclerosis at various sites of the vascular system, different risk factors reveal a varying system, different risk factors reveal a varying impact, e.g. cholesterol on ischemic heart disease and stroke. Although we have previously shown an association of hsp antibodies and carotid atherosclerosis, no data were available about the impact of coronary heart disease (CHD) on hsp65 antibody titres. Therefore, it seemed to us of interest, whether an elevation of hsp65 antibodies is specific for carotid atherosclerosis or also occurs when other sites of the vascular system are affected. In order to study a possible correlation between hsp65 antibodies and CHD or myocardial infarction (MI), 203 patients with CHD and MI and 76 healthy probands were recruited and serum antibodies to hsp65 were determined. Furthermore, the hsp65 antibody titres were statistically evaluated for a correlation with other established cardiovascular risk factors.

Section snippets

Subjects

In this sex- and age-matched case-control study, 279 subjects from the area of Innsbruck, Tyrol, Austria, aged 60–80 years, were consecutively recruited by the Department of Internal Medicine, University of Innsbruck. Each subject attending our Department within a time period of 12 months (group I, II, III) and fulfilling the given criteria was included in this study. All healthy control probands, attending as out patients for a health check-up test, were without a history of angina, revealed

Results

Since many factors are believed to be involved in the development of atherosclerosis and in the induction of hsp65 antibodies, in the present study, several established risk factors for atherosclerosis were recorded from all subjects. Data summarized in Table 1 shows average levels of blood lipids, i.e. cholesterol, LDL, HDL, triglycerides, and blood pressure, body weight, smoking and alcohol intake. No significant difference of the blood lipid levels and other parameters was found among the

Discussion

The involvement of immunomechanisms in atherosclerosis 1, 2and the association of hsp antibodies with carotid atherosclerosis [3]support the hypothesis that atherosclerosis represents an immunologically influenced disease. Therefore immune and autoimmune processes targeting the strongly conserved and highly immunogenic hsp65 molecule, may be pivotal for the inflammatory process in atherogenesis.

Until now hsp65 has been shown to be involved in the pathogenesis of inflammatory and autoimmune

Acknowledgements

Supported by grants (projects No. P11615-MED and P10677) from the Austrian Science Foundation. Recombinant mycobacterial hsp65 was kindly provided by Dr. J.D.A. van Embden, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.

References (20)

G Wick et al.
Is atherosclerosis an immunologically mediated disease?
Immunol Today
(1995)
Q Xu et al.
Association of serum antibodies to heat-shock protein 65 with carotid atherosclerosis
Lancet
(1993)
MK Heng et al.
Heat-shock protein 65 and activated γ/δ T cells in injured arteries
Lancet
(1994)
J Frostegård et al.
Induction of heat shock protein in monocytic cells by oxidized low density lipoprotein
Atherosclerosis
(1996)
PCM Res et al.
Synovial fluid T cell reactivity against the 65 kD heat shock protein of mycobacteria in early chronic arthritis
Lancet
(1988)
A Knowlton
The role of heat shock proteins in the heart
J Mol Cell Cardiol
(1995)
DH Birnie et al.
Anti-heat shock protein 65 titer in acute myocardial infarction
Lancet
(1994)
P Libby et al.
Involvement of the immune system in human atherogenesis. Current knowledge and unanswered questions
Lab Invest
(1991)
G Schett et al.
Autoantibodies against heat shock protein 60 mediate endothelial cytotoxity
J Clin Invest
(1995)
Q Xu et al.
Staining of endothelial cells and macrophages in atherosclerotic lesions with human heat shock protein reactive antisera
Arterioscler Thromb
(1993)

There are more references available in the full text version of this article.

Cited by (119)

Heat shock proteins and cardiovascular disease
2014, Advances in Clinical Chemistry
Atherosclerosis is the leading global cause of mortality, morbidity, and disability. Heat shock proteins (HSPs) are a highly conserved family of proteins with diverse functions expressed by all cells exposed to environmental stress. Studies have reported that several HSPs may be potential risk markers of atherosclerosis and related cardiovascular diseases, or may be directly involved in the atherogenic process itself. HSPs are expressed by cells in atherosclerotic plaque and anti-HSP has been reported to be increased in patients with vascular disease. Autoimmune responses may be generated against antigens present within the atherosclerotic plaque, including HSP and may lead to a cycle of ongoing vascular injury. It has been suggested that by inducing a state of tolerance to these antigens, the atherogenic process may be limited and thus provide a potential therapeutic approach. It has been suggested that anti-HSPs are independent predictors of risk of vascular disease. In this review, we summarize the current understanding of HSP in cardiovascular disease and highlight their potential role as diagnostic agents and therapeutic targets.
The potential role of heat shock protein 27 in cardiovascular disease
2012, Clinica Chimica Acta
Citation Excerpt :
It has been reported that Hsp27 stimulates monocyte production of anti-inflammatory cytokines such as IL-10 [117] and also inhibits their expression of toll like receptor-4 (TLR-4) and their differentiation into dendritic cells. Wick et al. [108] have hypothesized that the autoimmune responses to Hsps could be crucial in the initiation of atherosclerosis [118,119] and this is supported by human studies [97,111,120–122]. In a rat model in which aortic atherosclerosis is induced by a high cholesterol diet, 46 proteins were differentially expressed in the aortic tissue; among these differences was a decreased level of phosphorylated Hsp-27 [123].
Heat shock proteins (Hsps) comprise several families of proteins expressed by a number of cell types following exposure to stressful environmental conditions that include heat, free radicals, toxins and ischemia, and are particularly involved in the recognition and renaturation of mis-folded proteins. Heat shock protein-27 (Hsp27) is a member of the small Hsp (sHsp) family with a molecular weight of approximately 27 KDa. In addition to its chaperoning functions, Hsp27 also appears to be involved in a diverse range of cellular functions, promoting cell survival through effects on the apoptotic pathway and plays important roles in cytoskeleton dynamics, cell differentiation and embryogenesis. Over the past two decades there has been an increasing interest in the relationship between Hsp27 and cardiovascular disease. Hsp27 is thought to exert an important role in the atherosclerotic process. Serum Hsp27 concentrations appear to be a biomarker of myocardial ischemia. In this review, we will focus on the possible protective and immuno-modulatory roles of Hsp27 in atherogenesis with special emphasis on their changes following acute coronary events and their potential as diagnostic and therapeutic targets.
A cross-sectional study of the association between heat shock protein 27 antibody titers, pro-oxidant-antioxidant balance and metabolic syndrome in patients with angiographically-defined coronary artery disease
2011, Clinical Biochemistry
To investigate the association between serum antibody titers to Hsp27 (anti-Hsp27) and pro-oxidant–antioxidant balance (PAB) in patients with angiographically-defined coronary artery disease (CAD) with or without the metabolic syndrome (MS).
Subjects (n = 243) were classified into MS+ (n = 161) and MS− (n = 82) subgroups, based on the AHA/NHBLI criteria.
Serum anti-Hsp27 titers were found to be significantly higher in the MS+ vs. MS− group. However, no significant difference was observed in serum PAB values. When assessed for individual components of MS, increased serum anti-Hsp27 was found to be higher in subgroups with elevated triglycerides, elevated blood pressure and reduced high-density lipoprotein cholesterol (HDL-C). Subgroups of patients with elevated triglycerides had higher PAB values. HDL-C was the only significant predictor of anti-Hsp27 in the population as a whole.
The evidence from this investigation indicates the presence of elevated anti-Hsp27 in patients with concurrent CAD and MS compared to those with CAD alone.
Hsp60 and heme oxygenase-1 (Hsp32) in acute myocardial infarction
2011, Translational Research
Heat shock proteins (Hsps) are produced in response to various stressors, including ischemia-reperfusion, and they can exit cells and reach the blood. In this pilot study, we determined serum levels of Hsp60 and heme-oxygenase-1 (HO-1; also named Hsp32) in subjects with acute myocardial infarction (AMI) to assess their clinical significance and potential prognostic value. We also performed a bioinformatics analysis of the 2 molecules in search of structural clues on the mechanism of their release from cells. We studied 40 patients consecutively admitted for AMI (male:female patient ratio = 20:20, mean age: 64 ± 13 years) and 40 matched controls. A blood sample was drawn for biochemical analyses within 24 h of symptoms onset, and Hsp60 and HO-1 concentrations were determined by enzyme-linked immunosorbent assay (ELISA). All patients were followed up for 6 months to register adverse post-AMI cardiovascular events. A multivariate analysis demonstrated that elevated Hsp60 (P = 0.0361), creatine phosphokinase-muscle brain (CK-MB) (P = 0.0446), and troponin (P = 0.0490) were predictive of post-AMI adverse events. In contrast, increased HO-1 showed a significant association with less severity of coronary artery diseases (P = 0.0223). These findings suggest that Hsp60 and HO-1 play distinct roles in the pathogenesis of AMI and subsequent AMI-related pathology. The possibility that these proteins differ in their roles and mechanisms of action in AMI and post-AMI pathology was supported also by the bioinformatics estimates of probability of their localization in various subcellular compartments. The results clear the way for subsequent investigation on the pathogenetic role and clinical significance of Hsp60 and HO-1 in AMI.
Heat-shock proteins in cardiovascular disease
2011, Advances in Clinical Chemistry
Heat-shock proteins (HSPs) belong to a group of highly conserved families of proteins expressed by all cells and organisms and their expression may be constitutive or inducible. They are generally considered as protective molecules against different types of stress and have numerous intracellular functions. Secretion or release of HSPs has also been described, and potential roles for extracellular HSPs reported. HSP expression is modulated by different stimuli involved in all steps of atherogenesis including oxidative stress, proteolytic aggression, or inflammation. Also, antibodies to HSPs may be used to monitor the response to different types of stress able to induce changes in HSP levels. In the present review, we will focus on the potential implication of HSPs in atherogenesis and discuss the limitations to the use of HSPs and anti-HSPs as biomarkers of atherothrombosis. HSPs could also be considered as potential therapeutic targets to reinforce vascular defenses and delay or avoid clinical complications associated with atherothrombosis.
Heat-shock protein 90: A novel autoantigen in human carotid atherosclerosis
2009, Atherosclerosis
Citation Excerpt :
The same investigators provided direct proof in LDL-receptor deficient mice that HSP65-specific lymphocytes and IgG can promote fatty-streak formation [41] that can be suppressed by oral tolerization with HSP65 [42]. Studies in humans described a humoral response to HSP65 in patients with carotid atherosclerosis or coronary artery disease [43,44]. Environmental stress was shown to upregulate HSP60 expression on membrane cell surfaces and trigger secretion of a soluble form [45].
The known role of heat-shock proteins (HSPs) in the pathogenesis of atherosclerosis prompted us to investigate whether HSP90 is a target autoantigen of immune responses in patients with carotid atherosclerosis.
The presence of HSP90 on 26 cryostat and 6 paraffin embedded sections of carotid atherosclerotic plaques was determined by immunohistochemistry and immunofluorescence. Plaque-infiltrating T lymphocytes from 9 patients and circulating PBMC from 26 patients and 21 healthy subjects were tested by cell proliferation assay and by flow cytometry and ELISA for cytokine production in response to HSP90. ELISA was used to detect soluble HSP90 and anti-HSP90 antibodies in serum samples. Strong HSP90 immunoreactivity was detected in the muscle and endothelial cell layer and in the inflammatory infiltrate of carotid plaques. Plaque-derived and circulating T lymphocytes from patients proliferated in response to HSP90 whereas cells from healthy subjects did not. HSP90-specific T lymphocytes expressed IFN-γ and IL-4 suggesting concomitant Th1 and Th2 activation. ELISA detected soluble HSP90 in 42% and anti-HSP90 antibodies in 46% of patients’ sera.
These new findings, showing that HSP90 is overexpressed in plaque and serum from patients with atherosclerosis and induces an immune response in these patients, implicate HSP90 as a possible target autoantigen in the pathogenesis of carotid atherosclerosis.

View all citing articles on Scopus

View full text

Changes of serum antibodies to heat-shock protein 65 in coronary heart disease and acute myocardial infarction

Abstract

Introduction

Section snippets

Subjects

Results

Discussion

Acknowledgements

Immunol Today

Lancet

Lancet

Atherosclerosis

Lancet

J Mol Cell Cardiol

Lancet

Involvement of the immune system in human atherogenesis. Current knowledge and unanswered questions

Lab Invest

Autoantibodies against heat shock protein 60 mediate endothelial cytotoxity

J Clin Invest

Staining of endothelial cells and macrophages in atherosclerotic lesions with human heat shock protein reactive antisera

Arterioscler Thromb