Kein Folientitel

1
Vaccination and Immune Modulation against Heat
Shock Proteins
Qingbo Xu, MD, PhD St. Georges Hospital Medical
School University of London, UK
The 1st Atherosclerosis Vaccine Symposium, New
Orleans, 2004
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HSP family
HSPs Involved in atherosclerosis HSP27 HSP
45 HSP60 HSP70 HSP90 -?
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E. coli Hsp60 (GroEL)

• Assembled in a complex of
• 7 subunits together
• with GroES
• Implicated in macromolecular
• assembly and prevention of
• protein misfolding
• Translocates proteins
• across membranes
• Epitopes of bacterial HSP
• are present on the surface
• of certain bacteria,
• e.g. H. pylori

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HSP Expression
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HuHSP60 expression in the vessel wall
Cell marker anit-HSP Ab
EC
SMC
M?
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HSP expression in lesions of apoE-/- mice

Kanwar et al, ATVB 2001
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ApoE-/- mouse (8 months)
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Heat shock transcription factor (HSF) activation
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Rac1 dependent-activation of HSF1 in SMCs
stimulated by mechanical stress
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Mechanical stress
?
?
PKC
?
Rac
Oxidases
O2-?
Enzyme?
HSF1
Gene expression, e.g. HSP
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Can HSPs from cells and bacteria release into
circulation? Soluble HSPs
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Distribution of sHSP60 serum levels
300
200
Number of subjects
100
0
0
1-100
gt1000
101-200
201-300
301-400
401-500
501-600
601-700
701-800
801-900
901-1000
sHSP60 concentrations (ng/ml)
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Serum sHSP60 levels and odds ratio of incident
atherosclerosis
p0.004

2.0
2.0
odds ratio of incident atherosclerosis
1.3
1.0
1.1
1.0
0.0
I/II III IV V
n331
n162
n166
n167
sHSP60 concentrations (I-V quintile)
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Isolated Serum sHSP60
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What is sHSP doing? -activates vascular
endothelial and SMCs -activates
macrophages -activates immune system
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HSPs activate NF-kB in endothelial cells
Kol et al, JCI 1999
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Heat-shock protein 60 acting as cytokine
Activation of human vascular endothelial cells to
produce

• E-selectin

• ICAM-1

• VCAM-1

• IL-6

Verdegaal et. al., J. Immunol., 1996 Kol et. al.,
J. Clin. Invest., 1999
Stimulation of macrophages to produce

• TNF-?

• MMPs

• NO

Kol et. al., Circulation, 1998
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CD14
TLR4
MD2
p38MAPK
MyD88 IRAK TRAF6
Ras/Raf
Pro-ILs ILs
IKKs
MEK
I ?B
ERK1/2
NF-?B
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Autoantibodies against HSPs
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Xu et al. Association of serum antibodies to
heat-shock protein 65 with carotid
atherosclerosis. Lancet. 1993 Jan
30341(8840)255-9.
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Correlation of anti-HSP antibodies
n40, r20.87, plt0.001
n40, r20.56, plt0.001
Anti-mHSP65 antibodies
10 100 1000 10000
10 100 1000 10000
10 100 1000 10000
Anti-GroEL antibodies (titers)
Anti-cHSP60 antibodies (titers)
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Crossreactivity of purified anti-HSP60
antibodies in Western blot analysis
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Anti-HSP65 Antibody Shares Cross-reactive Epitope
with Mouse Monoclonal Antibody
II-13
ATVB 2003231060-1065
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Antibody-complement mediated endothelial damage
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Mouse Anti-HSP60 Antibody II-13 Induces
Endothelial Damage in ApoE-deficient Mice
Control
LPS
II-13
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Human Anti-HSP65 Antibodies induce
Atherosclerosis In ApoE-deficient Mice


Control
Ig-low
Total Ig-high
Specific Ig-high
Plt0.05
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Mouse Anti-HSP60 Antibody II-13 induces
Atherosclerosis In ApoE-deficient Mice
Mouse Ig
II-13 (x2)
II-13
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T cells reactive to HSPs Xu et al. 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 Jun91(6)2693-702.
Mosorin et al. Detection of Chlamydia
pneumoniae-reactive T lymphocytes in human
atherosclerotic plaques of carotid artery.
Arterioscler Thromb Vasc Biol. 2000
Apr20(4)1061-7. Curry et al. T lymphocyte
lines isolated from atheromatous plaque contain
cells capable of responding to Chlamydia
antigens.Clin Exp Immunol. 2000 Aug121(2)261-9.
Choi et al. Establishment of Porphyromonas
gingivalis heat-shock-protein-specific T-cell
lines from atherosclerosis patients. J Dent Res.
2002 May81(5)344-8. Chung et al. Immune
responses to heat shock protein in Porphyromonas
gingivalis-infected periodontitis and
atherosclerosis patients. J Periodontal Res.
200338388-93.
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HSP60-Reactive CD4CD28null T Cells in Patients
With Acute Coronary Syndromes
Zal et al. Circulation, 20041091230 - 1235.
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HSP expression in the arterial wall and/or other
tissues
sHSPs
Apoptosis
Protection
T and B cells
TLR4/CD14
Atherosclerosis
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Summary Wick G, Kloflach M and Xu Q. Autoimmune
and Inflammatory Mechanisms in Atherosclerosis.
Ann. Rev. Immunol. 22361-403, 2004. Impact for
therapy or prevention? Maron et al. Mucosal
Administration of Heat Shock Protein-65 Decreases
Atherosclerosis and Inflammation in Aortic Arch
of Low-Density Lipoprotein Receptor-Deficient
Mice Circulation 2002 106 1708-1716. Hansson
GK. Vaccination Against Atherosclerosis Science
or Fiction? Circulation 2002 106
1599-1601. Vaccination using a peptide of HSP60?
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Oak Foundation
Collaborators at other labs
S. Keichl J. Willeit Bruneck study, University
of Innsbruck, Austria J.C. Kaski SGHMS,
London H. Markus SGHMS, London
Vascular Biology Lab
M. Jahangiri SGHMS, London G. Schett University
of Vienna, Austria G. Wick University of
Innsbruck, Austria
G. Fontenos K. Mandal A. Afzal A. Shaw X. Yin E.
Torsney L. Ly D. Lo Z. Zhang
U. Mayr Q. Xiao A. Zampetaki I. Yusuf M. Mayr Y.
Hu L. Zeng S. Jacob Q. Xu