Cytokine 66 (2014) 95100

Contents lists available at ScienceDirect

Cytokine
journal homepage: www.journals.elsevier.com/cytokine

Cytokines and hs-CRP levels in individuals treated with low-dose aspirin

for cardiovascular prevention: A population-based study (CoLaus Study)
Julien Vaucher a,, Pedro Marques-Vidal b, Grard Waeber a, Peter Vollenweider a
a
b

Department of Medicine, Internal Medicine, CHUV and Faculty of Biology and Medicine, Lausanne, Switzerland
Institute of Social and Preventive Medicine (IUMSP), CHUV and Faculty of Biology and Medicine, Lausanne, Switzerland

a r t i c l e

i n f o

Article history:
Received 12 March 2013
Received in revised form 8 October 2013
Accepted 13 January 2014

Keywords:
Cytokines
hs-CRP
Low-dose aspirin
Cardiovascular prevention
Population-based sample

a b s t r a c t
Pro-inammatory cytokines and high-sensitive C-reactive protein (hs-CRP) are associated with increased
risk for cardiovascular disease. Low-dose aspirin for CV prevention is reported to have anti-inammatory
effects. The aim of this study was to determine the association between pro-inammatory cytokines and
hs-CRP levels and low-dose aspirin use for cardiovascular prevention in a population-based cohort
(CoLaus Study). We assessed blood samples in 6085 participants (3201 women) aged 3575 years.
Medications use and indications were recorded. Among aspirin users (n = 1034; 17%), overall low-dose
users (351; 5.8%) and low-dose for cardiovascular prevention users (324; 5.3%) were selected for analysis.
Pro-inammatory cytokines (IL-1b, IL-6 and TNF-a were assessed by a multiplex particle-based ow
cytometric assay and hs-CRP by an immunometric assay. Cytokines and hs-CRP were presented in
quartiles. Multivariate analysis adjusting for sex, age, smoking status, body mass index, diabetes mellitus
and immunomodulatory drugs showed no association between cytokines and hs-CRP levels and low-dose
aspirin use for cardiovascular prevention, either comparing the topmost vs. the three other quartiles (OR
95% CI, 0.84 (0.591.18), 1.03 (0.781.32), 1.10 (0.831.46), 1.00 (0.671.69) for IL-1b, IL-6, TNF-a and
hs-CRP, respectively), or comparing the topmost quartile vs. the rst one (OR 95% CI, 0.87 (0.601.26),
1.19 (0.791.79), 1.26 (0.861.84), 1.06 (0.671.69)). Low-dose aspirin use for cardiovascular prevention
does not impact plasma pro-inammatory cytokine and hs-CRP levels in a population-based cohort.
2014 Elsevier Ltd. All rights reserved.

1. Introduction
During the past two decades, inammation has been recognized
as an independent risk factor for cardiovascular (CV) diseases
[14]. In this context, pro-inammatory cytokines and C-reactive
protein (CRP) levels have been shown to be associated with
atherosclerosis [4] as well as heart failure [5], diabetes [6] and obesity [7].
The main mechanism of action of low-dose aspirin for primary
or secondary CV prevention is attributed to its anti-platelet aggregation activity through the inhibition of thromboxane A2 (TXA2)
via acetylation of cyclooxygenase (COX)-1 [8,9]. Besides inhibition
of platelets activity, low-dose aspirin has been postulated to present an anti-inammatory effect in patients with ischemic heart
disease or with type 2 diabetes without CV disease by reducing
the levels of the pro-inammatory cytokines [1014]. Although
the impact of statins on inammation has been extensively studied
[3,4,15], the effect of low-dose aspirin remains controversial and
insufciently investigated [4,16].
Corresponding author. Address: Etude CoLaus, Btiment des Instituts, 19, rue du
Bugnon, 1005 Lausanne, Switzerland. Tel.: +41 21 314 03 46; fax: +41 21 314 80 37.
E-mail address: julien.vaucher@chuv.ch (J. Vaucher).
1043-4666/$ - see front matter 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.cyto.2014.01.003

In a previous publication using data from the CoLaus Study, we

established population-based levels for several inammatory
markers such as interleukin (IL)-1b, IL-6 and tumor necrosis factor
(TNF)-a and for high-sensitive CRP (hs-CRP) [7]. The aim of the
present study was to determine the association between low-dose
aspirin for primary or secondary CV prevention and circulating levels of these pro-inammatory cytokines and hs-CRP, using data
from the same study.
2. Materials and methods
2.1. Participants
The study was approved by the Institutional Ethics Committee
of the University of Lausanne. The design of the CoLaus Study
has already been described [7,17]. Concisely, Lausanne (Canton
de Vaud, Switzerland) inhabitants were invited to participate to
the study. The following inclusion criteria were applied: (a) age
3575 years; (b) written informed consent; (c) willingness to
participate in the examination and to donate blood sample; and
(d) Caucasian origin. The CoLaus Study included only Caucasians
to reduce heterogeneity for genetic analyses. Caucasians extraction
was dened as having both Caucasian parents and grandparents.

96

J. Vaucher et al. / Cytokine 66 (2014) 95100

No other exclusion criteria were applied. Recruitment took place

between June 2003 and May 2006 and enrolled 6188 participants
(3251 women); participation rate was 41%. All participants attended the outpatient clinic of the University Hospital of Lausanne
in the morning after an overnight fast. Data were collected by
trained eld interviewers in a single visit lasting about 60 min.
Self-administered questionnaires, mailed with the appointment
letter, were brought by the participants and reviewed during the
visit.
2.2. Clinical data
Body weight and height were measured with participants
standing without shoes in light indoor clothes. Body weight was
measured in kilograms to the nearest 0.1 kg and height was measured to the nearest 5 mm. Body mass index (BMI) was dened
as weight/height2. Obesity was dened as BMI P 30 kg/m2 and
overweight as BMI P 25 kg/m2 and <30 kg/m2.
Smoking status was recorded in a self-administered questionnaire, collected data including the previous and current smoking
status as well as age of beginning and end (for former smokers).
Diabetes mellitus was dened as a fasting blood glucose P7 mmol/
l and/or oral antidiabetic and/or insulin treatment.
Personal history of CVD was dened as self-reported diagnosis
of coronary heart disease (myocardial infarction, angina pectoris,
and coronary revascularization), stroke or peripheral arterial
disease.
2.3. High-sensitivity CRP and cytokines measurements
Venous blood samples (50 mL) were drawn after an overnight
fast and allowed to clot. Hs-CRP was assessed by immunoassay
and latex HS (Immulite 1000-High, Diagnostic Products Corporation, LA, CA, USA) with maximum intra- and inter-batch coefcients
of variation of 1.3% and 4.6%, respectively. Serum samples were kept
at 80 C before assessment of IL-1b, IL-6, and TNF-a using a multiplex particle-based ow cytometric assay (Milliplex, Millipore, Zug,
Switzerland). The analysis was then conducted by a conventional
ow cytometer (FC500 MPL, BeckmanCoulter, Nyon, Switzerland).
This method is well correlated with standard procedures (ELISA)
[1820]. Lower limits of detection (LOD) for IL-1b, IL-6 and TNF-a
were 0.2 pg/ml. A good agreement between signal and cytokine
was found within the assay range (R2 P 0.99). Intra and inter-assay
coefcients of variation were respectively 15% and 16.7% for IL-1b,
16.9% and 16.1% for IL-6 and 12.5% and 13.5% for TNF-a. For quality
control, repeated measurements were conducted in 80 subjects,
randomly drawn from the initial sample, with a good reproducibility as described by Marques-Vidal et al. [7].
2.4. Medication use
All currently used medications were recorded, by asking the
participants to bring to the visit all those they were taking on a
regular basis during the 6 previous months or more. Their precise
dosages were not recorded but distinction was made between
over-the-counter drugs and those prescribed by a physician.
Moreover, daily and occasional use was also recorded. Finally,
medications indications were based on participants reports.
For the analysis, only drugs prescribed by a physician were
taken into account. The denition of low-dose aspirin users was
based on the association of 2 parameters: (1) daily use; (2)
indication for which low-dose aspirin is recommended according
to European guidelines in cardiology: primary or secondary CV
prevention, atrial brillation, prosthetic heart valves and peripheral arterial disease [2124]. Low-dose aspirin users for primary
or secondary CV prevention were further highlighted for analytical

purpose. Finally, it has to be considered that the dosage of aspirin

for primary or secondary cardiovascular prevention in Switzerland
ranges between 100 and 300 mg/d only.
Occasional aspirin users and/or those taking aspirin for another
indication than those listed above, thus called mix-group users,
were excluded from the analysis. The justication was double:
(1) heterogeneity of this group, mainly in terms of dosages; (2)
to focus on the aim of this study to determine if low-dose aspirin
for primary or secondary CV prevention has an impact on inammatory markers.
Statins, non-steroidal anti-inammatory drugs (NSAIDs), antibiotics, steroids and immunosuppressants use, which can inuence
inammatory parameters, were also collected.
2.5. Statistical analysis
Statistical analysis was conducted using STATA v11.0 (Stata
Corp, College Station, Texas, USA). Results were expressed as number of participants (percentage), as mean standard deviation and
as median and interquartile range (IQR). Bivariate analyses were
conducted using chi-square, Students t-test or KruskallWallis
non-parametric test. As previously reported, a signicant proportion of cytokine measurements, namely IL-1b, were below the
LOD [7]. Hence, for the association analysis, it was decided to analyse cytokines and hs-CRP in quartiles, and to include undetectable
values (below LOD) in the rst quartile. Multivariate analysis was
performed using logistic regression adjusting for sex, age, smoking
status, BMI, diabetes mellitus, concomitant statin, NSAIDs, antibiotics, steroids or immunosuppressants use. Two models were
tested: (a) the rst one assessed the likelihood of being in the topmost vs. the three other quartiles of cytokine distribution and (b)
the second one assessed the likelihood of being in the highest vs.
the lowest quartile. The effect of aspirin use in overall aspirin users,
overall low-dose aspirin users and low-dose aspirin users for CV
prevention was also assessed. Results were expressed as odds ratio
(OR) and 95% condence interval (CI). Statistical signicance was
considered for p < 0.05.

3. Results
Of the 6188 initial participants, 6085 (98.3%) could be assessed
for inammatory biomarkers while for the remaining 103 subjects
no blood samples were available. The analysis comprised 3201
women and 2884 men.
3.1. Characteristics associated with aspirin use
Among the 6085 participants included in the analysis, there
were 1034 (17%) aspirin users, of which 581 (56.2%) were men.
Among aspirin users, 351 (33.9%) took a low-dose regimen
(<300 mg/d) for a clear cardiovascular indication; 324 (31.3%) took
aspirin specically for primary or secondary cardiovascular prevention. 683 (66.1%) took high-dose aspirin (>300 mg/d) and/or took
aspirin for another indication than cardiovascular. Participants
characteristics, e.g. CV risk factors, according to aspirin use are
summarized in Table 1. Participants characteristics for statin,
non-steroidal anti-inammatory drugs (NSAIDs), antibiotics and
immunosuppressants use according to aspirin use category are
available in an online supplementary table.
CV risk factors (age, high BMI, former or current smoker,
diabetes mellitus) were all associated with the likelihood of being
treated with low-dose aspirin (p < 0.001). Statin use was associated
with low-dose aspirin use (p < 0.001) but not NSAIDs, antibiotics,
steroids nor immunosuppressants.

97

J. Vaucher et al. / Cytokine 66 (2014) 95100

Table 1
Participants characteristics according to aspirin use category.
Characteristics

Non-aspirin
users

Overall aspirin users

Mix-group
users

Low-dose users (overall)

Low-dose users for CV prevention

Overall population
(n = 6.085)
Demographic factors
Age
Means (years, SD)
Groups (n)
3545 years
4555 years
5565 years
6575 years
Gender
Men
Women
Smoking status
Never
Former
Current
BMI (kg/m2)
Means (SD)
Classication (n)
<25
2530
>30
Diabetes
Yes
No

5051 (83%)

1034 (17%)

683 (11.2%)

351 (5.8%)

324 (5.3%)

52.7 (10.6)

55.1 (11.3)

51.2 (10.6)

62.7 (8.6)

62.9 (8.5)

1315 (26.0)
1509 (29.9)
1363 (27.0)
864 (17.1)

226
250
310
248

(21.9)
(24.2)
(30.0)
(23.9)

214 (31.3)
206 (30.2)
178 (26.1)
85 (12.4)

12 (3.4)
44 (12.5)
132 (37.6)
163 (46.5)

12 (3.7)
36 (11.1)
123 (38)
153 (47.2)

2303 (45.6)
2748 (54.4)

581 (56.2)
453 (43.8)

364(53.3)
319 (46.7)

217 (61.8)
134 (38.2

207 (63.9)
117 (36.1)

2093 (41.4)
1604 (31.8)
1354 (26.8)

352 (34.0)
405 (39.2)
277 (26.8)

256 (37.5)
238 (34.9)
189 (27.7)

96 (27.4)
167 (47.6)
88 (25)

84 (25.9)
157 (48.5)
83 (25.6)

25.7 (4.4)

26.6 (4.9)

25.8 (4.5)

28.3 (5.2)

28.5 (5.2)

2502 (49.5)
1804 (35.7)
745 (14.8)

423 (40.9)
418 (40.4)
193 (18.7)

331 (48.5)
264 (38.6)
88 (12.9)

92 (26.2)
154 (43.9)
105 (29.9)

79 (24.4)
142 (43.8)
103 (31.8)

267 (5.2)
4784 (94.8)

120 (11.6)
914 (88.4)

30 (4.4)
653 (96.6)

90 (25.6)
261 (74.4)

90 (27.8)
234 (72.2)

pValue

<0.001
<0.001

<0.001

<0.001

<0.001
<0.001

<0.001

Characteristics of the study population (n = 6.058) according to the use of aspirin. Individuals who were on a low-dose regimen were selected: low-dose aspirin (max. 300 mg/
d) denition was based on the association of 2 parameters: (1) daily basis use and (2) indication for which low-dose aspirin is recommended according to international
guidelines in cardiology (primary or secondary CV prevention, atrial brillation, biological prosthetic heart valves and peripheral arterial disease). Low-dose aspirin users for
primary or secondary CV prevention were further highlighted for analytical purpose. Occasional aspirin users and/or those taking aspirin for another indication than those
listed above are presented in the mix-group users column. Results are expressed as number of participants (percentage) and as mean standard deviation. CV: cardiovascular
(primary or secondary). SD: standard deviation.

3.2. Association between aspirin use, plasma cytokine and hs-CRP

levels
On bivariate analyses, low-dose aspirin users for CV prevention
presented higher circulating IL-6 and TNF-a levels than non-users
(Table 2); no signicant differences were found for IL-1b and hsCRP.
Multivariate analysis of the association between aspirin use and
cytokine and hs-CRP quartiles was conducted using logistic regression adjusting for sex, age, smoking status, BMI, diabetes mellitus,
concomitant statin, NSAIDs, antibiotics, steroids or immunosuppressants use. The results are summarized in Table 2. The rst
analysis (top) assessed the likelihood of being in the topmost vs.
the three other quartiles of cytokine distribution. No signicant
association was found comparing overall aspirin users to
non-users; no further association was found for overall low-dose
aspirin users and low-dose aspirin users for CV prevention. Similar
results were obtained in the second analysis (top2), which assessed
the likelihood of being in the topmost vs. the lowest quartile. A
further adjustment with alternative exclusion of selected variables
(statins, NSAIDs, antibiotics, steroids and immunosuppressants)
led to similar results (not shown).

4. Discussion
Association between inammation and CV risks factors and diseases represents nowadays a fertile eld of investigations [4]. Concerning the role of low-dose aspirin on inammatory markers, and
consequently on CV disease evolution, a denite statement cannot
be made on actual evidence. Indeed, several studies dealing with
this topic have presented contradictory results [1014,2527]. In
this context, this large population-based study failed to show any
impact of low-dose aspirin for primary or secondary cardiovascular

prevention on circulating levels of IL-1b, IL-6, TNF-a and hs-CRP;

conrming previous results of smaller studies [2527].
Besides platelets COX-1 inhibition, aspirin exerts anti-inammatory effects by blocking COX-2, which promotes the conversion
of arachidonic acid to 15R-hydroxyeicosatetraenoic acid. The latter
is further transformed into various bioactive lipid mediators, called
aspirin-triggered lipoxins (ATLs), which act as local anti-inammatory mediators [28]. ATLs have been recognized as inhibitors of TNFa and IL-1b release by human polymorphonuclear leukocytes
[29,30]. Moreover, aspirin and its metabolite salicylate have been
shown to inhibit intracellular signaling pathways, of which nuclear
factor (NF)-jB and activator protein 1 (AP-1) [13,31,32]. These transcription factors play a central role in cellular responses to inammatory stimuli: activation of NF-jB results, for example, in the
production of chemokines which in turn attract immune cells. The
latter secrete pro-inammatory cytokines, such as IL-1, IL-6 and
TNF-a [33]. AP-1 is also implicated in the regulation of the expression of many cytokine genes [34,35]. Conversely, TNF-a activates
NF-jB and AP-1 pathways [36]. Still, most of the in vitro studies designed to investigate anti-inammatory effects of aspirin on these
pathways used higher dosages than those applied in clinical practice as demonstrated by Morris et al. [37]. Moreover, Bhagat et al.
have demonstrated that only high-dose aspirin (1 g) has an impact
on cytokines activity [38]. Thus, although the association between
atherosclerosis and inammation is clearly established, the impact
of anti-inammatory treatments to prevent the occurrence or the
progression of atherosclerosis remains doubtful [32,39]. Our results
seem to conrm the lack of effect of low-dose aspirin on circulating
levels of pro-inammatory cytokines and hs-CRP, which are implicated in the pathophysiology of atherosclerosis.
Nevertheless, various studies have concluded that, besides its
anti-aggregative activity, low-dose aspirin could have an antiinammatory effect which may contribute to the diminution of
cardiovascular risk [16,26,40]. However, although well designed,

98

J. Vaucher et al. / Cytokine 66 (2014) 95100

Table 2
Median and interquartile ranges of IL-1b, IL-6, TNF-a and hs-CRP. Results of multivariate analysis assessing association between aspirin use category and IL-1b, IL-6, TNF-a and
hs-CRP levels.
IL-1b (pg/ml)

IL-6 (pg/ml)

TNF-a (pg/ml)

hs-CRP (ng/ml)

0.40 (0.101.74)
0.38 (0.101.76)
0.744

1.28 (0.583.18)
1.54 (0.613.46)
0.028

2.85 (1.774.48)
3.03 (1.944.76)
0.006

1.3 (0.62.7)
1.4 (0.72.9)
0.099

0.40 (0.101.74)
0.27 (0.12.5)
0.23 (0.11.23)
0.744

1.28 (0.583.18)
1.28 (0.692.96)
2.08 (0.973.79)
0.028

2.85 (1.774.48)
3.5 (2.116.43)
3.39 (2.295.05)
0.006

1.3 (0.62.7)
1.0 (0.81.6)
1.7 (0.94)
0.099

1293 (21.3%)

1524 (25.1%)

1513 (24.9%)

1571 (25.8%)

1070 (82.8)
223 (17.2)
58 (4.5)
51 (3.9)

1246 (82.4)
278 (18.2)
100 (6.5)
95 (6.2)

1235 (81.6)
278 (18.4)
107 (7.1)
98 (6.5)

1290 (82.1)
281 (17.9)
112 (7.1)
109 (6.9)

OR for top
Aspirin
Non-users
Overall users
Low-dose (overall)
Low-dose for CV prevention prevention

1 (Ref.)
1.10 (0.931.3)
0.89 (0.641.24)
0.84 (0.591.18)

1 (Ref.)
1.04 (0.891.23)
1.00 (0.761.32)
1.03 (0.781.38)

1 (Ref.)
1.09 (0.931.28)
1.14 (0.871.49)
1.10 (0.831.46)

1 (Ref.)
0.99 (0.841.17)
0.93 (0.71.23)
1.00 (0.751.33)

OR for top2
Aspirin
Non-users
Overall users
Low-dose (overall)
Low-dose for CV prevention

1 (Ref.)
1.19 (0.981.44)
0.91 (0.641.29)
0.87 (0.601.26)

1 (Ref.)
0.98 (0.81.2)
1.10 (0.751.62)
1.19 (0.791.79)

1 (Ref.)
1.17 (0.961.43)
1.32 (0.911.9)
1.26 (0.861.84)

1 (Ref.)
1.02 (0.81.29)
0.96 (0.611.51)
1.06 (0.671.69)

Median and IQR

Aspirin
Non-users
Overall users
p-Value
Aspirin
Non-users
Low-dose (overall)
Low-dose for CV prevention
p-Value
Participants in the topmost quartile (%)
Overall (n = 6085)
Aspirin
Non-users
Overall users
Low-dose (overall)
Low-dose for CV prevention prevention

Results are expressed as median and interquartile range (IQR) of measured values, percentage (%) of participants in the topmost quartile and odds ratios of the two
multivariate analyses for IL-1b,IL-6, TNF-a and hs-CRP. The rst multivariate analysis (top) compares the topmost vs. the three other quartiles of cytokine distribution
adjusting for sex, age, smoking status, BMI, diabetes mellitus, concomitant statin, NSAIDs, antibiotics, steroids or immunosuppressants use. The second analysis (top2)
assessed the likelihood of being in the highest vs. the lowest quartile adjusting for the same variables. IL-1b: Interleukin-1b .IL-6: Interleukin-6. TNF-a: Tumor necrosis factora. hs-CRP: high-sensitive C-reactive protein. CV: cardiovascular.

these studies lacked statistical power. On the contrary, the present

study is population-based and included more than 6000 participants, with more than 300 subjects taking aspirin for CV prevention. Furthermore, our results are in agreement with other
studies where low-dose aspirin had no impact on CRP levels in
healthy individuals [2527].
We performed a post hoc calculation based on the results obtained and using a 5% signicance level and an 80% power showed
that a total number of 38,630 participants would be required to
achieve a statistically signicant odds ratio for aspirin users
relative to non-users for IL-1b; the corresponding values for IL-6
and TNFa are 168,080 and 35,250 respectively. Overall, these results suggest that the effect of aspirin on circulating cytokines levels, if any, has probably only marginal clinical and public health
impact.
Higher levels of cytokines have been previously demonstrated
in patients with stable angina or following acute myocardial infarction compared to control subjects [41]. On bivariate analyses, we
also found higher circulating IL-6 and TNF-a levels in low-dose
aspirin users compared to non-users. This can reect the inammatory state accompanying CV risk factors and diseases as previously published [7]. At least, this nding reinforces the
hypothesis that low-dose aspirin plays probably no signicant role
on inammatory mediators.
This study has some limitations. Participation rate was low
(41%), which might limit the external validity of the results; however, this participation rate is similar to other epidemiological studies undertaken in Europe [42]. The precise dosage of aspirin was not
collected, but only daily and occasional use as well as the indication

for which the drug was taken. However, dosages of low-dose aspirin
preparations for cardiovascular prevention in Switzerland range between 100 and 300 mg/d, which is in agreement with international
guidelines [2124]. Adherence to aspirin treatment was not evaluated and a low compliance could have led to a lack of impact on
cytokines and hs-CRP levels. Nevertheless, as this study has a population-based design, our results represent the reality encountered in
clinical practice. Moreover our results are similar to other studies
[16]. Other drugs might have inuenced inammatory state, such
as antibiotics, steroids or immunosuppressants. Few subjects were
on these regimens and alternative exclusion of them in the analysis
showed no impact on the results. Moreover, aspirin users could have
had higher baseline values of inammatory markers, which were
not recorded in this cross-sectional study. Then, aspirin might have
reduced cytokines and hs-CRP circulating levels but not at the level
as that of non-users. Finally, studied cytokines (IL-1b, IL-6, TNF-a
are not the sole mediators of inammation implicated in the pathogenesis of atherosclerosis: among others, VCAM-1, soluble CD40 ligand, adiponectin, IL-18, matrix metalloproteinase 9 and NO are
also implicated [37,41]. However, IL-1b, IL-6 and TNF-a are recognized as central mediators of inammation in atherosclerosis via
the activation of various pathways, which are modulated by aspirin
[28,43,44].
In summary, no association between low-dose aspirin use and
circulating levels of pro-inammatory cytokines and hs-CRP was
found using a large, population-based study. The effects of lowdose aspirin appear to be related mainly to its well-known antithrombotic activity rather than to its putative anti-inammatory
effects.

J. Vaucher et al. / Cytokine 66 (2014) 95100

Contributorship
 Pedro-Marques Vidal and Peter Vollenweider have contributed
to the analysis and interpretation of data; they have also revised
critically the article.
 Grard Waeber has revised critically the article.
 Julien Vaucher has contributed to the conception and design of
the study; to the analysis and interpretation of data; and to
draft the article.
Funding

[15]

[16]

[17]

[18]
[19]

This work was supported by grants from the Swiss National Science Foundation [Grant No.: 33CSCO-122661]; GlaxoSmithKline;
the Faculty of Biology and Medicine of Lausanne, Switzerland;
and the Swiss Society of Internal Medicine.

[20]

[21]

Acknowledgments
The authors also express their gratitude to the participants in
the Lausanne CoLaus study and to the investigators who have contributed to the recruitment, in particular Yolande Barreau, AnneLise Bastian, Binasa Ramic, Martine Moranville, Martine Baumer,
Marcy Sagette, Jeanne Ecoffey and Sylvie Mermoud for data
collection.
Appendix A. Supplementary material

[22]

[23]

[24]

Supplementary data associated with this article can be found, in

the online version, at http://dx.doi.org/10.1016/j.cyto.2014.01.003.
References
[1] Libby P, Hansson GK. Involvement of the immune system in human
atherogenesis: current knowledge and unanswered questions. Lab Invest
1991;64:515.
[2] Ridker PM, Hennekens CH, Buring JE, Rifai N. C-reactive protein and other
markers of inammation in the prediction of cardiovascular disease in women.
N Engl J Med 2000;342:83643.
[3] Ridker PM, Danielson E, Fonseca FA, Genest J, Gotto Jr AM, Kastelein JJ, et al.
Rosuvastatin to prevent vascular events in men and women with elevated Creactive protein. N Engl J Med 2008;359:2195207.
[4] Libby P, Ridker PM, Hansson GK. Inammation in atherosclerosis: from
pathophysiology to practice. J Am Coll Cardiol 2009;54:212938.
[5] Vasan RS, Sullivan LM, Roubenoff R, Dinarello CA, Harris T, Benjamin EJ, et al.
Inammatory markers and risk of heart failure in elderly subjects without
prior myocardial infarction: the Framingham heart study. Circulation
2003;107:148691.
[6] Pickup JC. Inammation and activated innate immunity in the pathogenesis of
type 2 diabetes. Diabetes Care 2004;27:81323.
[7] Marques-Vidal P, Bochud M, Bastardot F, Luscher T, Ferrero F, Gaspoz JM, et al.
Levels and determinants of inammatory biomarkers in a Swiss populationbased sample (CoLaus study). PLoS One 2011;6:e21002.
[8] Collaborative meta-analysis of randomised trials of antiplatelet therapy for
prevention of death, myocardial infarction, and stroke in high risk patients.
BMJ. vol. 324; 2002. p. 7186.
[9] Baigent C, Blackwell L, Collins R, Emberson J, Godwin J, Peto R, et al. Aspirin in
the primary and secondary prevention of vascular disease: collaborative metaanalysis of individual participant data from randomised trials. Lancet
2009;373:184960.
[10] Ikonomidis I, Andreotti F, Economou E, Stefanadis C, Toutouzas P,
Nihoyannopoulos P. Increased proinammatory cytokines in patients with
chronic stable angina and their reduction by aspirin. Circulation
1999;100:7938.
[11] Chen YG, Xu F, Zhang Y, Ji QS, Sun Y, Lu RJ, et al. Effect of aspirin plus
clopidogrel on inammatory markers in patients with non-ST-segment
elevation acute coronary syndrome. Chin Med J (Engl) 2006;119:326.
[12] Hovens MM, Snoep JD, Groeneveld Y, Frolich M, Tamsma JT, Huisman MV.
Effects of aspirin on serum C-reactive protein and interleukin-6 levels in
patients with type 2 diabetes without cardiovascular disease: a randomized
placebo-controlled crossover trial. Diabetes Obes Metab 2008;10:66874.
[13] Muhlestein JB. Effect of antiplatelet therapy on inammatory markers in
atherothrombotic patients. Thromb Haemost 2010;103:7182.
[14] Gao XR, Adhikari CM, Peng LY, Guo XG, Zhai YS, He XY, et al. Efcacy of
different doses of aspirin in decreasing blood levels of inammatory markers

[25]

[26]

[27]

[28]
[29]

[30]

[31]
[32]
[33]

[34]

[35]

[36]

[37]

[38]
[39]
[40]

99

in patients with cardiovascular metabolic syndrome. J Pharm Pharmacol

2009;61:150510.
Glynn RJ, Koenig W, Nordestgaard BG, Shepherd J, Ridker PM. Rosuvastatin for
primary prevention in older persons with elevated C-reactive protein and low
to average low-density lipoprotein cholesterol levels: exploratory analysis of a
randomized trial. Ann Int Med 2010;152(48896):W174.
Solheim S, Pettersen AA, Arnesen H, Seljeot I. No difference in the effects of
clopidogrel and aspirin on inammatory markers in patients with coronary
heart disease. Thromb Haemost 2006;96:6604.
Firmann M, Mayor V, Vidal PM, Bochud M, Pecoud A, Hayoz D, et al. The
CoLaus study: a population-based study to investigate the epidemiology and
genetic determinants of cardiovascular risk factors and metabolic syndrome.
BMC Cardiovasc Disord 2008;8:6.
Vignali DA. Multiplexed particle-based ow cytometric assays. J Immunol
Methods 2000;243:24355.
Dupont NC, Wang K, Wadhwa PD, Culhane JF, Nelson EL. Validation and
comparison of luminex multiplex cytokine analysis kits with ELISA:
determinations of a panel of nine cytokines in clinical sample culture
supernatants. J Reprod Immunol 2005;66:17591.
Kellar KL, Mahmutovic AJ, Bandyopadhyay K. Multiplexed microsphere-based
ow cytometric immunoassays. Curr Protoc Cytom; 2006 [Chapter 13: Unit13
1].
Graham I, Atar D, Borch-Johnsen K, Boysen G, Burell G, Cifkova R, et al.
European guidelines on cardiovascular disease prevention in clinical practice:
full text. Fourth Joint Task Force of the European Society of Cardiology and
other societies on cardiovascular disease prevention in clinical practice
(constituted by representatives of nine societies and by invited experts). Eur
J Cardiovasc Prev Rehabil 2007;14 Suppl 2:S1113.
Camm AJ, Kirchhof P, Lip GY, Schotten U, Savelieva I, Ernst S, et al. Guidelines
for the management of atrial brillation: the Task Force for the Management
of Atrial Fibrillation of the European Society of Cardiology (ESC). Eur Heart J
2010;31:2369429.
Vahanian A, Baumgartner H, Bax J, Butchart E, Dion R, Filippatos G, et al.
Guidelines on the management of valvular heart disease: the task force on the
management of valvular heart disease of the European society of cardiology.
Eur Heart J 2007;28:23068.
Tendera M, Aboyans V, Bartelink ML, Baumgartner I, Clement D, Collet JP, et al.
ESC Guidelines on the diagnosis and treatment of peripheral artery diseases:
Document covering atherosclerotic disease of extracranial carotid and
vertebral, mesenteric, renal, upper and lower extremity arteries: the Task
Force on the Diagnosis and Treatment of Peripheral Artery Diseases of the
European Society of Cardiology (ESC). Eur Heart J 2011;32:2851906.
Feng D, Tracy RP, Lipinska I, Murillo J, McKenna C, Toer GH. Effect of shortterm aspirin use on C-reactive protein. J Thromb Thrombolysis
2000;9:3741.
Feldman M, Jialal I, Devaraj S, Cryer B. Effects of low-dose aspirin on serum Creactive protein and thromboxane B2 concentrations: a placebo-controlled
study using a highly sensitive C-reactive protein assay. J Am Coll Cardiol
2001;37:203641.
Kim MA, Kim CJ, Seo JB, Chung WY, Kim SH, Zo JH, et al. The effect of aspirin on
C-reactive protein in hypertensive patients. Clin Exp Hypertens
2011;33:4752.
Spite M, Serhan CN. Novel lipid mediators promote resolution of acute
inammation: impact of aspirin and statins. Circ Res 2010;107:117084.
Clish CB, OBrien JA, Gronert K, Stahl GL, Petasis NA, Serhan CN. Local and
systemic delivery of a stable aspirin-triggered lipoxin prevents neutrophil
recruitment in vivo. Proc Natl Acad Sci USA 1999;96:824752.
Hachicha M, Pouliot M, Petasis NA, Serhan CN. Lipoxin (LX)A4 and aspirintriggered 15-epi-LXA4 inhibit tumor necrosis factor 1alpha-initiated
neutrophil responses and trafcking: regulators of a cytokine-chemokine
axis. J Exp Med 1999;189:192330.
Kopp E, Ghosh S. Inhibition of NF-kappa B by sodium salicylate and aspirin.
Science 1994;265:9569.
Amann R, Peskar BA. Anti-inammatory effects of aspirin and sodium
salicylate. Eur J Pharmacol 2002;447:19.
Bollrath J, Greten FR. IKK/NF-kappaB and STAT3 pathways: central signalling
hubs in inammation-mediated tumour promotion and metastasis. EMBO Rep
2009;10:13149.
Zenz R, Eferl R, Scheinecker C, Redlich K, Smolen J, Schonthaler HB, et al.
Activator protein 1 (Fos/Jun) functions in inammatory bone and skin disease.
Arthritis Res Ther 2008;10:201.
Smolinska MJ, Page TH, Urbaniak AM, Mutch BE, Horwood NJ. Hck tyrosine
kinase regulates TLR4-induced TNF and IL-6 production via AP-1. J Immunol
2011;187:604351.
Chung F, Yegneswaran B, Liao P, Chung SA, Vairavanathan S, Islam S, et al. STOP
questionnaire: a tool to screen patients for obstructive sleep apnea.
Anesthesiology 2008;108:81221.
Morris T, Stables M, Hobbs A, de Souza P, Colville-Nash P, Warner T, et al.
Effects of low-dose aspirin on acute inammatory responses in humans. J
Immunol 2009;183:208996.
Bhagat K, Vallance P. Inammatory cytokines impair endothelium-dependent
dilatation in human veins in vivo. Circulation 1997;96:30427.
Libby P, Ridker PM, Hansson GK. Progress and challenges in translating the
biology of atherosclerosis. Nature 2011;473:31725.
Balbay Y, Tikiz H, Baptiste RJ, Ayaz S, Sasmaz H, Korkmaz S. Circulating
interleukin-1 beta, interleukin-6, tumor necrosis factor-alpha, and soluble

100

J. Vaucher et al. / Cytokine 66 (2014) 95100

ICAM-1 in patients with chronic stable angina and myocardial infarction.

Angiology 2001;52:10914.
[41] Solheim S, Arnesen H, Eikvar L, Hurlen M, Seljeot I. Inuence of aspirin on
inammatory markers in patients after acute myocardial infarction. Am J
Cardiol 2003;92:8435.
[42] Tolonen H, Koponen P, Aroma A, Conti S, Graff-Iversen S, Grtvedt L, Kanieff M,
Mindell J, Natunen S, Primatesta P, Verschuren M, Viet L, Kuulasmaa K. for the

Feasibility of a European Health Examination Survey (FEHES) Project. Review

of Health Examination Surveys in Europe. 2008;Helsinki, Finland: National
Public Health Institute (KTL). Report No.: B18/2008; 2008.
[43] Packard RR, Libby P. Inammation in atherosclerosis: from vascular biology to
biomarker discovery and risk prediction. Clin Chem 2008;54:2438.
[44] Frantz S, Ertl G, Bauersachs J. Mechanisms of disease: toll-like receptors in
cardiovascular disease. Nat Clin Pract Cardiovasc Med 2007;4:44454.