SHAPE Society

1. Editorial Slides
VP Watch, February 19, 2003, Volume 3, Issue 6
What Is The Link Between Smoking
And Plaque Inflammation?

2. –Furie et al Habitual use of smokeless
tobacco leads to accumulation of
inflammatory leukocytes at the site of
placement, which may contribute to
tissue damage. 2
–Their observations suggested that
smokeless tobacco may induce
inflammatory changes in vivo by
activating endothelium in a manner that
promotes recruitment of leukocytes.2

3. –Tottie et al showed that
nicotine is chemotactic for
neutrophil and enhances
neutrophil responsiveness to
chemotactic peptides. 3
–Klapproth et al showed nicotine
and ligands of nicotine
receptors release GM-CSF from
epithelial cells.4

4. – Bobryshev and Lord have
reported “Vascular Dendritic
Cells” in areas prone to
atherosclerotic plaque
formation. 5
- These dendritic cells may
play a key role in the
initiation of atherosclerosis. 6

5. –As featured in VP Watch of the
Week, Aicher et al1
investigated the
effect of nicotine on the dendritic
cell- mediated adaptive immunity.
–In the following slides, the
investigators showed various
effectd of nicotine on expression of
nicotinic acetylcholine receptor in
human dendritic cells.

6. αα7-nicotinic acetylcholine receptor (nAChR)7-nicotinic acetylcholine receptor (nAChR)
expression on DCs is up-regulated afterexpression on DCs is up-regulated after
nicotine stimulationnicotine stimulation
Control
αα7-nAChR expression7-nAChR expression
NAChR expression in DC cells was increased by nicotine in aNAChR expression in DC cells was increased by nicotine in a
dose-dependent manner.dose-dependent manner.
Nicotine: 10-8
M
Nicotine: 10-7
M
Provided by Dr. Stefanie Dimmeler

7. Nicotine induces a dose-dependent increase in the
expression of the co-stimulatory molecule CD86
PBS 10-8
10-7
10-6
10-5
10-4
10-7
10-4
M
ChangeinCD86expression(%)
Mecamylamine
(MEC; 10-7
M)
*
*
*
* *
* P<0.01 versus PBS
Nicotine
250
200
150
100
50
CD86
PBS
nicotine
isotype
MECMEC
Ca++Na+
K+
AChACh
NicotineNicotine
--
nAChRnAChR++
Provided by Dr. Stefanie Dimmeler

8. Dose-dependent cytotoxicity of nicotine
PercentageofdeadDCs
* P<0.01 versus PBS *
*
*
0
20
40
60
80
100
120
PBS 10-7
10-6
10-5
10-4
10-3
10-2
Nicotine
M
Provided by Dr. Stefanie Dimmeler

9. Nicotine enhances expression of surface
molecules involved in inflammation
Changeinexpression(%ofcontrol)
isotype
PBS
nicotine
PBS
nicotineisotype
PBS
nicotineisotype
PBS
nicotine
isotype
50
100
150
200
250
CD11aCD11a
CD18CD18
CD54CD54
CD40CD40
CD83CD83
CD86CD86
HLA-DRHLA-DR
CD86 CD54
MHC class II CD40
Provided by Dr. Stefanie Dimmeler

10. Nicotine-induced IL-12 p40 production in DCs is
mediated through nACh receptors
* P<0.01 versus PBS
PBS
IL-12(pg/ml)
200
100
0
Nicotine Nicotine
+
BTX
Nicotine
+
MEC
*
BTX: α-Bungarotoxin
(α7-nAChR antagonist)
MEC: Mecamylamine
(unselective nAChR
antagonist)
MECMEC
Ca++Na+
K+
AChACh
NicotineNicotine
--
nAChRnAChR
++BTX
Both nicotine antagonists
prevented nicotine-induced IL-12 P40 production Provided by Dr. Stefanie Dimmeler

11. Nicotine-prestimulated dendritic cells (DCs) and
monocytes (MCs) induce allogeneic T cell activation
IL-2(pg/ml)
nicotine
DC / T cell
ratio (1:10)
LPS
DC / T cell
ratio (1:50)
200
100
0
300
PBS
*
*
* *
* P<0.01 versus PBS
0
200
100
300
*
*
*
*
MC / T cell
ratio (1:10)
MC / T cell
ratio (1:50)
* P<0.01 versus PBS
IL-2(pg/ml)
nicotine
LPS
PBS
nicotine
LPS
PBS
nicotine
LPS
PBS
Provided by Dr. Stefanie Dimmeler

12. CSFE
CD4
MCs
control
nicotine
DCs
Proliferation of allogeneic CSFE-labeled CD4+ T cells in
mixed lymphocyte reactions with nicotine-stimulated
DCs or MCs as stimulator cells
CD4-allophycocyanin
7.2%
24.1%
5.2%
10.7%
Loss of incorporated CSFE labeling was increased by
nicotine, indicating its enhancing effect on proliferation
of T-lymphocytes. Provided by Dr. Stefanie Dimmeler

13. IL-2(pg/ml)
0
20
40
60
80
100
120
140
PBS 0.1 1.0
PBS Nicotine
*
*
*
OVA (µg/ml)
* P<0.01 versus PBS
Nicotine-preactivated DCs stimulate OVA-TCR-transgenic
T cells in OVA-antigen specific assays
OVA: ovalbumine
peptide 323-339
OVA-TCR
transgenic mice
OVA
OVA-
TCR
Provided by Dr. Stefanie Dimmeler

14. Nicotine-preactivated DCs increase expression
of CD40 ligand (CD40L) on T cells
0
10
20
30
40
PBS nicotine
CD40L+
Tcells(%)
*
* P<0.01 versus PBS
Provided by Dr. Stefanie Dimmeler

15. Nicotine activates MAPK and Akt pathways
phospho p38
β-tubulin
A
phospho Erk1/2
β-tubulin
B
0 5 15 30 60 120 min
phospho Akt
β-tubulin
C
These studies demonstrate that the effects of nicotine are mediated, at least
in part, by phosphorylation of Akt and MAPK
Provided by Dr. Stefanie Dimmeler

16. isotype
PBS
nicotine + PD98059
nicotine
nicotine + SB203580
nicotine + LY294002
CD86
LY294002: PI3-K inhibitor
PD98059: MEK 1/2 inhibitor
(ERK-pathway)
SB203580: p38 MAPK inhibitor
Nicotine-induced up-regulation of CD86 is strongly dependent
on MAPK and phosphatidylinositoI-3 (PI3) kinase
These studies demonstrate that the effects of nicotine are mediated, at least
in part, by phosphorylation of Akt and MAPK Provided by Dr. Stefanie Dimmeler

17. control nicotine
B
Homing of CSFE-labeled DCs to atherosclerotic plaques
in nicotine-treated hypercholesterolemic mice
Nicotine induced recruitment of dendritic cells into the
atherosclerotic plaque in vivo Provided by Dr. Stefanie Dimmeler

18. Summary
Nicotine enhances adaptive immunity and
may contribute to plaque destabilization
NicotineNicotine
AdaptiveAdaptive
immunityimmunity
PlaquePlaque
destabilizationdestabilization
Provided by Dr. Stefanie Dimmeler

19. Conclusion:
Nicotine has a direct
proinflammatory effect
through enhancement
of cell-mediated
adaptive immunity.

20. Conclusion:
Nicotine enhances the
recruitment of dendritic
cells to atherosclerotic
plaques which can result in
increasing recruitment of
monocytes / macrophages
to plaque.

21. Questions:
The role of smoking in cardiovascular disease is
more related to the contribution of nicotine:
1- to endothelial injury and initiation
of atherosclerotic plaque
2- to progression of plaques that
already exist
3- to thrombotic complications of
plaques
continue next page

22. Questions:
4- to increased thrombogenesity of
blood
5- to increased arrhythmogenesity of
myocardium
6- to increased respiratory infections
and total infectious burden