SHAPE Scientific Symposium, Dallas, TX 2013
Concept of a Vaccine for Vascular Disease
Modulating Autoimmunity in Atherosclerosis
Using a Vaccine
Kuang-Yuh Chyu, MD, PhD
Associate Director, CICU
Cedars Sinai Heart Institute
Cedars-Sinai Medical Center
Disclosures: Co-Inventor of ApoB-100 Peptide Vaccines for Atherosclerosis.
Patent rights are assigned to Cedars-Sinai Medical Center.
1
Atherosclerosis is an Auto-Immune Mediated Inflammatory Disease
Pro-atherogenic Adaptive Immunity
APC
Inflammatory
Cytokines
IFN- γ and TNF-α
Th1
Auto
antigen
NKT
MHC/CD1
Naïve
T cell
Th 17
Inflammatory
Cytokine
IL 17
Atherosclerosis
Activated CD4+ T-cell
2
Athero-protective Adaptive Immune Response
Antigen
Treg
Suppression of
Autoimmune Response
APC
Naive
T cell
Tolerogenic
Dendritic Cell
Regulatory T cell
CD25+, Foxp3+
Secrete anti-inflammatory
cytokines
IL-10/TGF-β
Th1
Athero-protective
Effect
3
Oxidized LDL in Human Coronary Plaques
Colocalizing with Macrophages
Auto-antigens
in
Atherosclerosis
LDL/Ox-LDL
Hsp-60
2GP1
OxLDL
Macrophages
4
Would Immunization with
LDL/oxLDL
Worsen or Reduce
Atherosclerosis?
5
Immunization of Cholesterol-fed Rabbits with Homologous LDL
Reduces Aortic Atherosclerosis Despite Hypercholesterolemia
5
4.5
4
3.5
3
2.5
P<0.05
2
1.5
1
0.5
0
Cholesterol
Control
N=7
Immunized
N=9
1259 mg/dl
1181 mg/dl
Ameli, Shah, Nillson et al: ATVB 1996
Nilsson, Ameli, Shah et al: JACC 1997
6
Effect of Immunization with Native and Oxidized
Homologous LDL On Experimental
Atherosclerosis in Hypercholesterolemic Mice
Immunization of LDL-R null mice with homologous native or
MDA-LDL reduces atherosclerosis (Freigang et al ATVB
1998)
Immunization of apo E null mice with homologous MDA-LDL
reduces atherosclerosis (George et al : Atherosclerosis
1998)
Timing affects the athero-protective effects of LDLimmunization (Chyu, Shah et al. Atherosclerosis 2004)
7
Apo B-100
550kD
4536 AA
302 Peptides, 20 amino acids long with 5 amino
acid overlap simulating the entire amino acid
sequence of human Apo B 100 were
synthesized.
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
Amino acid 3136-3155
KTTKQ SFDLS VKAQY KKNKH
p210
Johs, A. et al. JBC 2006;281:19732-19739
Apo B100 Related Peptide (p210) Vaccine
Reduces Atherosclerosis in Hyperlipidemic Mice
P < 0 .0 5
61%
10
5
p2
10
-c
B
cB
SA
SA
-A
/A
lu
lu
m
m
S
0
PB
% A o r tic lesion
15
P210: apo B-100 amino acids 3136-3155
Chyu KY, Shah PK et al: PLoS One 2012
9
Apo B100 Related Peptide (p210) Vaccine Reduces
Plaque Macrophages in ApoE-/- Mice
* p<0.05
PBS
Control
Adjuvant
Control
p210
Vaccine
Chyu KY, Shah PK et al: PLoS One 2012
Apo B100 Related Peptide (p210) Vaccine
Reduces Plaque Dendritic Cells in ApoE-/- Mice
p < 0.05
PBS
Control
Adjuvant
Control
p210
Vaccine
Chyu KY, Shah PK et al: PLoS One 2012
Adoptive Transfer of CD8 T-cells from p210
Immunized Mice Conveys Athero-protection
to Unimmunized Mice
p < 0.05
Chyu KY et al : PLoS One 2012
12
Adoptive Transfer of p210 loaded Dendritic
Cells reduces Atherosclerosis in Apo E-/- Mice
Nasal delivery of p210-CTB Fusion Construct
reduces Atherosclerosis in Apo E-/- Mice
Prolonged subcutaneous delivery of p210
reduces Atherosclerosis in Apo E-/- Mice
Abdominal aortic aneurysm formation after
AngII infusion in apoE-/- mice
Daugherty et al Br J Pharmacol. 2001 October; 134(4): 865–870.
14
% Total Aneurysmal Segment
number of aneurysmal segment/number of total segment (%)
(Aneurysmal segment = diameter>1.35mm)
1
2
3
p<0.05
4
6
5
8
7
Number of aneurysmal segment (severity of disease) was
significantly reduced in p210 immunized mice.
Honjo T: Circulation (abst) 2011
p210 Vaccine Reduces Mortality in AngiotensinII
Induced Aneurysm Model in ApoE-/- Mice
Survival Curve
Mortality
P210 = 6.2%
PBS = 32.1%
p<0.05 vs. PBS
p<0.05 vs. cBSA
cBSA = 29.7%
by Logrank test
Honjo T: Circulation (abst) 2011
16
p210 Vaccine Reduces Pro-inflammatory IL-17 Positive CD4 T cells
CD4+IL-17A+
*
*
p < 0.05
N: PBS=8, cBSA=6, p210=6
Honjo T et al : unpublished data 2013
Systolic BP
180
mmHg
160
P<0.05
P210
140
cBSA
120
PBS
AngII
100
7w
8w
9w
10w
11w
12w
13w
week
Mean BP
170
mmHg
150
P210
130
cBSA
110
P<0.05
PBS
AngII
90
7w
8w
9w
10w 11w
week
12w
13w
Honjo T: Circulation (abst) 2011
18
P210 Vaccine Reduces AT1R Expression in Aorta
IB; AT1R
IB; GAPDH
N=6, P<0.05
PBS
cBSA
p210
p210 vaccine reduced the expression of AT1R in whole aorta.
Honjo T: Circulation (abst) 2011
19
Conclusions
• Immunization with selected ApoB-100 peptides
reduces atherosclerosis and favorably modifies plaque
phenotype in hypercholesterolemic mice.
• Immunization with selected ApoB-100 peptides also
reduced AngII induced hypertension and aneurysm
formation and mortality due to rupture.
• These experimental observations provide proof of
concept regarding the feasibility and efficacy of apoB100 peptide based vaccine for vascular diseases.
20
Lund University, Sweden
Oppenheimer Atherosclerosis Research Center
P.K.Shah, MD
Kuang-Yuh Chyu, MD, PhD
Paul Dimayuga,PhD
Tomoyuki Honjo, MD,PhD
Xiaoning Zhao,PhD
Jianchang Zhou, PhD
Nicole Lio
Bojan Cercek, MD, PhD
Cheryl Chan
Juliana Yano
Cardiovax
Jan Nilsson, MD, PhD
GN Fredriksson,PhD
Funding
NIH-NHLBI
Eisner Foundation
Cardiovax
The HeartFoundation
Spielberg Foundation
Corday Foundation
21
Potential Mechanisms of Action of p210 Vaccine
Increased Lysis of
APC
p210
Suppression
of
Antigen Specific
adverse
Autoimmune Response
APC
CD8+T cell
CD8+ Regulatory T cell
CD25+, IL-10+
22
IL-17 Deletion Protects Mice from AngiotensinII
Induced Aortic Aneurysm Formation
Survival
Incidence
46.7%
0%
Honjo T et al : unpublished data 2013
Abdominal Aortic Aneurysm
Pathophysiology
Inflammation
Immune activation
Oxidative stress
Matrix proteolysis
SMC death