Download Immunomodulation of Atherosclerosis

AEHA-AHA-Nov 12-2005, Dallas
“Immunomodulation of Atherosclerosis”
P.K.Shah, MD
Director, Division of Cardiology and
Atherosclerosis Research Center
Cedars Sinai Medical Center, Los Angeles
Vaccine for Atherosclerosis
“Vaccines for infectious diseases
are likely to be the most important
medical contribution to public health
during the last 100 years -------------”
Nilsson J , Hansson G K , Shah PK: ATVB 2004; 25: 1-11
CSMC--PKS
Yin and the Yang of Immune System in Atherosclerosis
Innate Immunity
Adaptive Immunity
Macrophages
Dendritic Cells
CRP
Toll like receptors
(TLR)
CSMC--PKS
Natural
Antibody
Scavenger Receptors
(SR-A, CD 36)
T-cells
B-cells
Immune Activation in Atherosclerosis
Both innate and adaptive immune responses modulate atherosclerosis
Auto-antigens
Hsp-60:
2GP1 :
ox-LDL:
Consequences of
Immune Response
Pro-atherogenic
Pro-atherogenic
???
Immune Response to Oxidized /MDA-LDL
Apo B100
Apo B100
Cholesterol
Cholesterol
Phospholipid
Phospholipid
Plaque
Formation
LDL cholesterol
Oxidized LDL
Phospholipid
Neoantigens
Apo B 100
Neoantigens
Immune Recognition
Macrophage
CSMC--PKS
B-cells
T-cells
(antibodies)
(cytokines)
Immunization of Cholesterol-fed Rabbits with Homologous LDL Substantially
Reduces Aortic Atherosclerosis Despite Hypercholesterolemia
Ameli, Shah, Nillson et al :ATVB 1996
Nilsson , Ameli, Shah et al: JACC 1997
Apo B100
Apo B100
Cholesterol
Cholesterol
Phospholipid
LDL Cholesterol
Control
N=7
Cholesterol
CSMC--PKS
1259mg/dl
Immunized
N=9
1181mg/dl
Phospholipid
Oxidized LDL
-Antigen: 280 mcg LDL
-Adjuvant: 700 mcg AdjuPrime
-Primary SC Vaccination followed by 1 booster
-Animals euthanzied 16 weeks after vaccination
Immunization of LDL-Receptor Deficient (Watanabe Rabbits) with
Homologous Malondialdehyde (MDA) Modified LDL Reduces Atherogenesis
Palinski , Witztum et al : PNAS 1995
% of Aortic Surface with Plaque
70
P<0.005
60
50
40
30
20
10
0
CSMC--PKS
Control
Rabbits
(N=11)
MDA-LDL
Immunized Rabbits
(N=14)
Apo B100
302 Peptides, 20 amino acids long with 5 amino acid
overlap simulating the entire amino acid sequence of
human Apo B 100 were synthesized.
Cholesterol
Phospholipid
Using an ELISA with peptides sequences as antigens,
antibodies to 101 of these peptide sequences were
identified in pooled human sera
Several peptide sequences were then used to create vaccines
for Immunization in apo E null mice fed a high
cholesterol diet
LDL Cholesterol
( Collaborative Research Program between
Cedars Sinai Medical Center (P.K.Shah) and
University of Lund (Jan Nilsson) )
Hypothesis: Specific antigenic epitopes on Apo B 100 component of
LDL provoke athero-protective immune response
CSMC--PKS
ATVB 2003
Peptide 143 + Peptide 210 Mixture
Immunization of Apo E Null Mice with Apo B-100 related
Peptide Sequence Reduces Atherosclerosis
Alum used as adjuvant
Mouse Apo B 100
Homology
EEEMLENVSLVCPKDATRFK
Peptide 1
ATRFKHLRKYTYNYQAQSSS
Peptide 2
6-7 wks
Ist vaccination
CSMC--PKS
8-9 wks
Booster
25 wks
Sacrifice
75%
85%
Immunization of Apo E Null Mice with Apo B-100 related Peptide Sequence :
Effect on Cholesterol Levels and Aortic Atherosclerosis
Serum cholesterol mg/dl
% of Aortic Surface Covered by Plaque
1600
3.5
1400
3
1200
2.5
1000
2
800
P<0.01
1.5
600
1
400
0.5
200
N=9
N=10
Alum
(Control)
Peptide 1
N=10
0
Peptide 2
Immunization Group
CSMC--PKS
0
N=9
N=10
N=10
Alum
(Control)
Peptide 1
Peptide 2
Immunization Group
Immunization of Apo E Null Mice with Apo B-100 related
Peptide Sequence Reduces Atherosclerosis
CSMC--PKS
Immunization of Apo E Null Mice with Apo B-100 related
Peptide Sequence Reduces Plaque Inflammation and Increases Collagen Content
% Collagen content (Trichrome)
% Macrophage immunoreactivity
14
p<0.05
45
40
12
35
10
30
8
25
20
6
p<0.05
4
2
N=9
0
Alum
(Control)
10
5
N=10
Peptide 1
Immunization Group
CSMC--PKS
15
N=10
Peptide 2
0
N=9
Alum
(Control)
N=10
Peptide 1
N=10
Peptide 2
Immunization Group
Late Immunization of Apo E Null Mice with Apo B-100 related
Peptide Sequence Attenuates Progression of Atherosclerosis
% Aortic Surface with Plaque
14
P<0.05
12
10
8
6
4
2
0
Immunization Group:
16 wk
30 wk
Alum Ctl
Cholesterol (mg/dl): 1274
930
16 wk
30 wk
Peptide 2
1274
989
Adoptive Transfer of Splenocytes from Peptide 2 Immunized Mice Reduces
Atherosclerosis in Recipient Unimmunized Apo E Null Mice
% of Aortic Surface Covered by Plaque
3.5
P<0.01
3
2.5
2
1.5
1
0.5
N=9
N=9
N=9
0
Mice receiving
Splenocytes
From Alum
Immunized mice
CSMC--PKS
Mice receiving
Splenocytes
From Peptide 1
Immunized mice
Mice receiving
Splenocytes
From Peptide 2
Immunized mice
Multiple Apo B-100 Related Peptide Antigens Have
Athero-protective Effects in Apo E Null mice
Fredrickson, Shah, Nilsson et al : ATVB 2003
% Aortic Atherosclerosis
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
Control
Peptide 45 Peptide 74
Peptide
210
Peptide
240
Peptides
11, 25,74
Peptides
30-34
Peptides
143,210
Conclusions
-Immune system plays a complex role in atherosclerosis with pro-atherogenic
and athero-protective effects
-Immunization using LDL/ox-LDL and specific Apo B-100 related peptide
sequences reduces atherosclerosis and favorably modifies plaque composition
- Immunotherapy of atherosclerosis warrants further investigation
Acknowledgements
Kuang-Yuh Chyu , MD, PhD (Cedars Sinai)
Xiaoning Li, PhD(Cedars Sinai)
Juliana Yano,BS (Cedars-Sinai)
Gunilla Nordin-Fredrickson, MD, PhD (Sweden)
Jan Nilsson, MD, PhD (Sweden)
Michael and Jane Eisner Foundation
CSMC--PKS