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Eur J Appl Physiol (2014) 114:2607–2616
DOI 10.1007/s00421-014-2979-6
ORIGINAL ARTICLE
Acute leukocyte, cytokine and adipocytokine responses
to maximal and hypertrophic resistance exercise bouts
Johanna Ihalainen · Simon Walker · Gøran Paulsen ·
Keijo Häkkinen · William J. Kraemer · Mari Hämäläinen ·
Katriina Vuolteenaho · Eeva Moilanen · Antti A Mero
Received: 24 March 2014 / Accepted: 5 August 2014 / Published online: 22 August 2014
© Springer-Verlag Berlin Heidelberg 2014
for adipocytokine profile. Immediate mechanical stress
seemed similar as no differences in myoglobin response
were observed. The higher magnitude of metabolic demand
reflected in higher lactate response in HYP could be the
reason for the significantly high responses in WBC, IL-1ra
and decrease in MCP-1.
Keywords Resistance exercise · White blood cells ·
Cytokines · Adipocytokines
A list of abbreviations
HYP Hypertrophic resistance exercise bout
IL-1ra Interleukin-1 receptor antagonist
IL-6 Interleukin-6
MAX Maximal resistance exercise bout
MCP-1 Monocyte chemoattractant protein-1
WBC White blood cell count
Introduction
Resistance training is associated with a reduced risk of
low-grade inflammation and improvement in metabolic
diseases such as cardiovascular disease and type 2 diabe-
tes (Kraemer et al. 2002; Gordon et al. 2009; Calle and
Fernandez 2010). Resistance training has been associated
with improvements in inflammation state in overweight
adults (Olson et al. 2007), elderly (Phillips et al. 2012) as
well as in specific patient groups (Conraads et al. 2002;
Moraes et al. 2014). A bout of heavy resistance exercise
triggers a transient inflammatory response comprising aug-
mented white blood cell count and stimulation of pro- and
anti-inflammatory cytokines production (Freidenreich and
Volek 2012). The physiological stress caused by heavy
resistance exercise acts as a major stimulus for muscle fiber
Abstract The purpose of this study was to examine
the acute immune response (circulating levels of leuko-
cytes, cytokines and adipocytokines) to maximal resist-
ance (MAX, 15 × 1RM) and hypertrophic resistance
(HYP, 5 × 10RM) exercise bouts. Twelve healthy men
(age = 28.2 ± 3.5 years, weight = 78.6 ± 10.4 kg, height
178.8 ± 5.0 cm, fat percentage = 16.5 ± 3.5 %) partici-
pated in the study. Blood was sampled before, immediately
after and 15 and 30 min after exercise. Leukocytes (WBC)
significantly increased immediately after HYP (p < 0.01),
whereas in MAX, increases in WBC became significant
after 30 min (p < 0.05). Lymphocytes increased only after
HYP (p < 0.001), while MAX induced lymphopenia during
recovery (p < 0.01). Monocyte chemoattractant protein-1
(MCP-1) decreased (p < 0.05) and interleukin-1 recep-
tor antagonist (IL-1ra) increased after HYP, which were
not observed after MAX. Adipsin and resistin decreased
after both exercise bouts (p < 0.05), which suggest that
heavy resistance exercise is at least transiently beneficial
Communicated by Fabio Fischetti.
J. Ihalainen (*) · S. Walker · K. Häkkinen · A. A. Mero
Department of Biology of Physical Activity, University
of Jyväskylä, P.O Box 35, 40014 Jyväskylä, Finland
e-mail: johanna.stenholm@jyu.fi
G. Paulsen
Norwegian School of Sport Sciences, Oslo, Norway
W. J. Kraemer
Human Performance Laboratory, Department of Kinesiology,
University of Connecticut, Storrs, USA
M. Hämäläinen · K. Vuolteenaho · E. Moilanen
The Immunopharmacology Research Group, University
of Tampere School of Medicine and Tampere University
Hospital, Tampere, Finland
2608 Eur J Appl Physiol (2014) 114:2607–2616
1 3
hypertrophy, and efficient repair of muscles requires a well-
coordinated and controlled inflammatory response (Peake
et al. 2010).
Well-known immunological responses after intensive
and prolonged endurance exercise include neutrophilia
(high neutrophil counts), lymphopenia (low lymphocyte
counts) and decreased cytotoxic activity in natural killer
cells (Nieman 1997). The inflammatory response to heavy
resistance exercise and especially after a traditional maxi-
mal resistance exercise, which is primarily designed to
maximize neural adaptation and places low metabolic
demands (few repetitions and long inter-set rest periods)
(Simonson and Jackson 2004; Hakkinen and Pakarinen
1993; Hulmi et al. 2012) is much less investigated (Paulsen
and Peake 2013). Resistance exercise may impact the mag-
nitude of circulating WBC in similar manner to endurance
exercise, but the exact mechanisms that induce responses
in the immune system during resistance exercise are not
known (Freidenreich and Volek 2012).
The cytokine response induced by a bout of heavy resist-
ance training involves enhanced production of pro-inflam-
matory cytokines, such as IL-1β and IL-1α and IL-6. In
addition to the pro-inflammatory properties, IL-6 has anti-
inflammatory effect as it stimulates the appearance of anti-
inflammatory cytokines, such as IL-1ra and IL-10. These
mediators play a crucial role in the containment and reso-
lution of inflammatory processes, and have been suggested
to link the acute effect of exercise to the decrease of car-
diovascular diseases associated with low-grade inflamma-
tion (Steensberg et al. 2003; Petersen and Pedersen 2005);
(Izquierdo et al. 2009) suggested that the magnitude of
metabolic demand and fatigue, which is experienced during
hypertrophic resistance exercise, affects both the cytokine
and hormone response patterns. Complementarily, Miles
et al. (2003) found evidence to support the hypothesis
that the increase in blood lactate levels may be part of the
mechanism to increase lymphocyte counts in the circula-
tion. Moreover, heavy resistance exercise also induces an
acute endocrine response, e.g., increased circulating levels
of cortisol and epinephrine (Kraemer et al. 1996; Nieman
et al. 1995b) which are known to be potential effectors for
the immune system (Pedersen and Hoffman-Goetz 2000).
One of the physiological changes responsible for the
positive effects of exercise on cardiovascular and metabolic
health has been suggested to be changes in adipocytokines
(Robinson and Graham 2004). Adipocytokines (leptin, adi-
ponectin, resistin and adipsin) are hormones that were first
discovered to be secreted by adipose tissue and to regulate
energy metabolism and appetite. More recent findings on
the ubiquitous expression of their receptors and on their
cellular effects have revealed that they are also involved in
the regulation of a variety of biological functions related to
immune responses and inflammatory diseases (Ouchi et al.
2011). A single exercise bout has been demonstrated to
exert specific acute effects on adipocytokine levels and the
response appears dependent on the duration of the exercise
and on energy expenditure (Bouassida et al. 2008).
Heavy resistance exercises for improving maximal
strength and muscle growth are important parts of a pro-
gressive resistance training program of athletes and could
be beneficial for the general population as well as for
elderly people with consistent resistance training experi-
ence. Therefore, it is important to characterize the imme-
diate effects of different types of resistance exercise on
blood WBC, cytokines and adipocytokines, and the pos-
sible mechanisms that link the type of resistance exer-
cise to the magnitude of the response. The purpose of the
present study was to examine the acute immune response
(changes in WBC, cytokines and adipocytokines) during
two heavy but specifically different resistance exercise
bouts: MAX (15 × 1RM) and HYP (5 × 10RM). Sec-
ondly, we investigated the relationship of muscle injury,
lactate accumulation and hormonal changes after the
exercise sessions. We hypothesized that, as the metabolic
demand is lower in maximal resistance type resistance
exercise, the acute inflammatory response (changes in
blood WBC counts and cytokine levels) would also be
lower after MAX than after HYP but beneficial changes
in adipocytokine profiles would be observed during
recovery after both loadings.
Methods
Participants
Twelve healthy, young males volunteered for this study
(age = 28.2 ± 3.5 years, weight = 78.6 ± 10.4 kg,
height 178.8 ± 5.0 cm, fat percentage = 16.5 ± 3.5 %;
mean ± SD). All participants reported taking part in sport
activities on a weekly basis, but none were competitive ath-
letes or had a background in systematic strength training.
Participants filled in a health questionnaire prior to par-
ticipation in the study. All subjects reported that they were
non-smokers, free from injury, and were not using any
medications. Each subject was informed of the potential
risks and discomforts associated with the measurements,
and all the subjects gave their written informed consent to
participate. The study was conducted according to the dec-
laration of Helsinki, and the ethics committee of the Uni-
versity of Jyväskylä, approved the study.
Pretesting
All subjects in this investigation participated in a familiari-
zation session, which included anthropometrics and body
2609Eur J Appl Physiol (2014) 114:2607–2616
1 3
composition measurement, as well as the one repetition
maximum (1RM) test performed in the leg press exercise
(David D210 leg press device, David Health Solutions
Ltd., Helsinki, Finland). This bilateral 1RM test was used
to determine the loads used in each acute exercise proto-
col. The starting position (flexed) was approximately 60°
for knee angle and 70° for hip angle, whereas the finishing
position, at full extension was 180° for the knee angle. As
part of their warm-up, the subjects performed 4 progressive
submaximal sets (1 set of 10 repetitions with 70 % of esti-
mated 1RM, 1 × 7 × 75 % estimated 1RM, 1 × 5 × 80 %
estimated 1RM, and 1 × 1 × 90 % estimated 1RM). After
the estimated submaximal sets, single repetitions were per-
formed with increasing loads (2.5–5 kg increments) until
the subject could not lift the load to the finishing position
of 180° knee angle. The rest period between attempts was
3 min. The last successful repetition (rep) was considered
to be the subject’s 1RM.
Experimental protocol
The first exercise session was performed 1 week after the
familiarization session. The order of the protocols was ran-
domized and counter-balanced. The MAX protocol was 15
sets of 1 repetition (reps) at 100 % of 1RM and the HYP
protocol was 5 sets of 10 reps at 80 % of 1RM for the leg
press exercise. The loads used during the first set were
determined from the 1RM load of the familiarization ses-
sion. The loads were adjusted during the sessions to enable
completion of the required reps. If the subject was not able
to complete the required reps, assistance was provided and
the load is reduced for the next set. The inter-set rest period
was 3 min for MAX and 2 min for HYP. The exercise bouts
were separated by 1 week.
Body composition
After an overnight fast, body composition body mass (BM),
total body muscle mass (MM), fat mass (FM) and per-
centage of body fat measurements were performed using
an eight-point bioelectrical impedance device (Inbody
720 body composition analyser, Biospace Co. Ltd, South
Korea.) The subjects were barefoot and wore shorts. Body
height was measured to the nearest 0.5 cm using a wall-
mounted scale.
Blood samples and analyses
Blood lactate was measured to determine the metabolic
effect of work performed in exercises. Blood samples were
obtained from the fingertip and collected into capillary
tubes (20 µL), which were placed in a 1-mL hemolyzing
solution and analyzed automatically after the completion of
testing according to the manufacturer’s instructions (EKF
diagnostic, C-line system, Biosen, Germany). The lactate
data has already been published previously (Walker et al.
2012).
Venous blood samples were drawn by repeated
venepunctures from an antecubital vein using standard
procedures. To assess the acute impact and short-term
recovery from exercise protocols, blood samples were col-
lected pre-exercise, immediately after (POST0), 15 min
after (POST15), and 30 min after (POST30) the exer-
cises (4 blood samples in total during each exercise bout
×10 mL/sample = 40-mL blood). Total (WBC) and dif-
ferential white blood cells, platelets, as well as hemoglobin
and hematocrit were determined from EDTA-treated blood
(Venosafe, Terumo, Belgium) with Sysmex KX-21 N
(TOA Medical Electronics Co., Ltd., Kobe, Japan). Of
WBC, neutrophils, lymphocytes and mixed cells (mono-
cytes, eosinophils, basophils and immature precursor cells)
were analyzed. In addition, venous blood was collected
into serum separator tubes (Venosafe, Terumo, Belgium).
The samples were centrifuged for 10 min at +4 °C with
2,000×g (Megafuge 1.0 R, Heraeus, Germany). Serum was
kept at −80 °C until analyzed for serum cortisol using the
Immulite 1,000 and hormone-specific immunoassay kits
(Immulite, Siemens, IL). Detection limit for cortisol was
5.5 nmoL/l and inter-assay coefficient of variation (CV %)
7.9 %. Concentrations of interleukin-6 (IL-6), monocyte
chemoattractant protein-1 (MCP-1), L-selectin (L-sel),
interleukin-1 receptor antagonist (IL-1ra), adipsin, adi-
ponectin, leptin, resistin and myoglobin in serum samples
were determined by enzyme-linked immunosorbent assay
(ELISA) with commercial reagents (IL-6: eBioscience,
San Diego, CA, USA; MCP-1, L-selectin, IL-1ra, adipsin,
adiponectin, leptin, and resistin: R&D Systems, Europe
Ltd, Abindgon, UK; myoglobin: USCN Life Science Inc.,
Wuhan, China). The detection limits and inter-assay coef-
ficients of variation, respectively, were 0.2 pg/mL and
3.4 % for IL-6, 3.9 pg/mL and 5.9 % for MCP-1, 19.5 pg/
mL and 2.9 % for L-selectin, 31.3 pg/mL and 7.6 % for IL-
1ra, and 0.78 ng/mL and 9.2 % for myoglobin, 15.6 pg/mL
and 4.5 % for adipsin, 31.3 pg/mL and 4.2 % for adiponec-
tin, 15.6 pg/mL and 5.8 % for resistin and 15.6 pg/mL and
3.8 % for leptin. Prior to the statistical analyses, because
of significant fluctuations in plasma in HYP, all data were
corrected for changes in plasma volume calculated using
hematocrit and hemoglobin described by Dill and Costill
(1974).
Statistical analysis
Data is presented as mean ± SE. Before applying further
statistical methods, the data were checked for sphericity and
normality. If a specific variable violated the assumptions of
2610 Eur J Appl Physiol (2014) 114:2607–2616
1 3
parametric tests then rank-transformation was used. Rank-
transformation was used for cortisol, IL-6, myoglobin, adi-
ponectin and leptin. Absolute and relative changes (e.g.,
WBC, acute cytokine and hormonal adaptations) were ana-
lyzed via two-way repeated analysis of variance for main
(type, time) and interaction (type × time) effects. This was
followed by one-way repeated measures ANOVA on each
MAX and HYP trials to examine a main effect of time. If
main or interaction was observed p ≤ 0.05 the change from
pre-values for POST0, POST15 and POST30 was compared
between type or time using paired t tests with Bonferroni
correction. Spearman’s rank correlation coefficient was
used to examine the relations. Data were analyzed using
PASW statistic 18.0 (SPSS, Chicago, IL, USA). The level of
statistical significance was set at p ≤ 0.05.
Results
Duration, total work, lactate, hormones and myoglobin
The duration of MAX was 50 min, whereas HYP
took 20 min to complete. Total work in MAX was
2,500 ± 380 kg and 7,240 ± 1,190 kg in HYP. There were
no significant differences between pre-exercise lactate, hor-
mone or myoglobin concentrations. Circulating levels of
lactate, cortisol and myoglobin are presented in Table 1.
There was a significant type × time (p < 0.01) and main
effect (p < 0.01) of time and type effect in lactate. The peak
lactate concentrations were observed at POST0 in MAX
(4.6 ± 0.6) and in HYP (12 ± 0.5) and the increase was
significant in both HYP (+670 %, p < 0.001) and in MAX
(+160 %, p < 0.01) at POST0 when compared to the pre-
exercise values, although the increase was significantly
higher (p < 0.001) in the HYP compared to the MAX pro-
tocol. Results for cortisol indicated significant type × time
interaction (p < 0.05) and a significant main effect for
type (p < 0.05) and time (p < 0.01). Post hoc compari-
son revealed that statistical differences between HYP and
MAX was significant at POST15 (p < 0.000) and POST30
(p < 0.01). Serum cortisol concentrations increased signifi-
cantly only after HYP (p < 0.01). There were no differences
in serum myoglobin levels between resistance exercise
bouts. Myoglobin concentrations increased significantly,
and at a similar magnitude in MAX (+182 %, p < 0.01)
and in HYP (+217 %, p < 0.001) protocols, and did not
significantly increase after POST0 but remained elevated
during the follow-up of 30 min.
White blood cells
There were no significant differences between pre-exercise
white blood cells or, platelet counts between exercise bouts.
White blood cell and subgroup counts and platelets are pre-
sented in Table 2. For the WBC a significant group × time
interaction (p < 0.05) was observed, and a significant main
effect for the type (p < 0.05) and time (p < 0.05) was found.
WBC (p < 0.001) significantly increased immediately
after HYP, whereas in MAX, increases in WBC counts
became significant only at POST30 (p < 0.05). A signifi-
cant group × time interaction (p < 0.05) was observed in
neutrophils and post hoc comparison revealed a signifi-
cant difference in acute increase on neutrophils (p < 0.05).
During recovery, neutrophil counts tended to decrease
from POST0 in HYP and were at basal levels at POST30.
Results for lymphocytes indicated a significant type × time
interaction (p < 0.05) and main effect of time (p < 0.01)
and type (p < 0.05). Lymphocyte responses were noticeably
different, as lymphocyte numbers increased immediately
after HYP and returned to pre-exercise levels during recov-
ery, whereas in MAX slight, but significant, decreases from
pre-exercise values were observed at POST15 (p < 0.05)
and POST30 (p < 0.05).
Cytokines, adipocytokines and L-selectin
There were no significant differences between pre-exer-
cise concentrations either in cytokines or adipocytokines
Table 1 Myoglobin, l-selectin, lactate and cortisol
Lactate was only measured at PRE, POST and POST15
MAX maximal, HYP hypertrophic
* significant difference to pre-exercise value
† significant difference between the exercise bouts. (mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, †p < 0.05, ††p < 0.01, †††p < 0.001)
PRE POST 0 POST 15 POST30
MAX HYP MAX HYP MAX HYP MAX HYP
Myoglobin (ng/mL) 23 (4.2) 25 (8.0) 44 (8.0)** 42 (6.5)*** 49 (8.0)** 50 (7.1)*** 51 (6.8)** 48 (4.4)***
L-selectin (ng/mL) 1,100 (16) 1,100 (14) 1,100 (16) 1,100 (18) 1,100 (17) 1,100 (20) 1,100 (15) 1,100 (19)
Lactate (mmol/L) 1.8 (0.2) 2.1 (0.1) 4.6 (0.6)** 12 (0.5)*** ††† 2.1 (0.2)* 11 (0.6)*** ††† – –
Cortisol (nmol/L) 290 (27) 300 (38) 300 (36) 330 (33)** †† 270 (33) 520 (24)** ††† 240 (30)* 600 (40)*** †††
2611Eur J Appl Physiol (2014) 114:2607–2616
1 3
between exercise bouts. For the IL-6 only significant main
effect of time (p < 0.05) was observed. IL-6 responses were
similar in both exercise protocols, as both showed a pro-
gressive increase after the exercise with the highest concen-
trations being observed at POST30 (+90 and +94 % for
MAX and HYP, respectively, Fig. 1a). A significant main
effect of time was observed in MCP-1. Interestingly, there
was a significant decrease (p < 0.01) in MCP-1 in HYP at
POST30 when compared to the PRE (Fig. 1b). A signifi-
cant type × time interaction (p < 0.05) was observed in
IL-1ra. In HYP, IL-1ra concentrations were significantly
increased at POST0 (p < 0.05, Fig. 1c) whereas in MAX
we did not observe significant differences. Significant
changes in serum L-selectin were not observed.
The effects of resistance exercises on adipocytokines are
presented in Fig. 2. For the adipsin response a significant
main effect for time (p < 0.01) was observed. Adipsin con-
centrations decreased immediately after MAX (p < 0.05)
and HYP (p < 0.01) and kept progressively decreasing
after both exercise protocols up to POST30. Adiponectin
and leptin levels were unaffected by exercises. A signifi-
cant main effect of time (p < 0.05) was observed in resis-
tin. Resistin decreased significantly (p < 0.01) in MAX and
HYP and progressively decreased during recovery.
Correlations
A negative correlation (r = −0.643, p < 0.05) was observed
between fat mass and adiponectin pre-exercise (Fig. 3a). In
HYP, a correlation was observed between myoglobin and
IL-1ra at POST0 (r = 0.701, p < 0.05, Fig. 3b) as well as
between myoglobin and neutrophils (r = 0.601, p < 0.05,
Fig. 3c). In MAX, there was a significant negative corre-
lation between the acute increase in WBC and the acute
increase in cortisol at POST0 (r = −0.622, p < 0.05,
Fig. 3d). Interestingly, the peak lactate (POST15) corre-
lated negatively with lymphocytes in HYP (r = −0.874,
p < 0.05, Fig. 3e) and in MAX (r = −0.697, p < 0.05,
Fig. 3f).
Discussion
This study demonstrated that both hypertrophic and maxi-
mal resistance exercise bouts alter immediate responses in
WBC and cytokine concentrations. Interestingly, the acute
increase in white blood cells was significantly higher after
HYP. The increase in total WBC was dominated by neutro-
phils in MAX, whereas in the HYP both neutrophils and
lymphocytes increased significantly. The higher metabolic
demand in HYP demonstrated by significantly higher lac-
tate response might explain the difference in lymphocyte
response. Both exercise bouts induced similar signs of
muscle injury, demonstrated by increased myoglobin con-
centrations. There was a significant decrease in lympho-
cytes during recovery at POST15 in MAX. IL-6 increased
significantly after both exercise sessions. Interestingly,
both exercise protocols decreased adipsin and resistin lev-
els immediately after and continuously during the 30 min
follow-up after the exercise. In addition, there was a sig-
nificant increase immediately after exercise in IL-1ra and
decrease at 30 min in MCP-1 in HYP. These observations
support the hypothesis that resistance exercise has benefi-
cial anti-inflammatory effects, which might be more pro-
nounced in HYP because the metabolic demand is higher
than during MAX. The findings of this study show the
immediate immune responses to MAX and HYP, however,
the evaluation is limited by the difference in duration of the
exercise bouts.
White blood cells
An increased mobilization of white blood cells into the cir-
culation was observed after both bouts. An acute increase
in blood WBC counts has been shown to occur after resist-
ance exercise (RE) in several studies (Nieman et al. 1995a;
Kraemer et al. 1996; Simonson 2001; Natale et al. 2003;
Ramel et al. 2003, 2004; Simonson and Jackson 2004;
Mayhew et al. 2005; Risoy et al. 2003). The type of resist-
ance exercise seems to affect the amplitude and time course
Table 2 White blood cells, subgroups, and platelets
MAX maximal, HYP hypertrophic, Mixed cells monocytes, eosinophils, basophils and immature precursor cells
* Significant difference to pre-exercise value
† Significant difference between the exercise bouts. (mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001, †p < 0.05, ††p < 0.01, †††p < 0.001)
PRE POST 0 POST 15 POST30
MAX HYP MAX HYP MAX HYP MAX HYP
Total WBC (×109/l) 6.4 (0.4) 7.0 (0.6) 7.1 (0.6) 9.9 (0.6)*** ††† 7.2(0.7) 9.2 (0.6)*** †† 7.6 (0.6)* 8.0 (0.5)*
Lymphocytes (×109/l) 2.2 (0.2) 2.3 (0.5) 2.2 (0.6) 3.8 (0.7)*** ††† 2.0 (0.5)* 3.4 (0.2)*** ††† 1.9 (0.5)* 2.4 (0.6)††
Neutrophils (×109/l) 3.6 (0.3) 4.0 (0.4) 4.1 (0.4) 5.1 (0.5)* † 4.6 (0.5)* 5.0 (0.4)* 4.9 (0.5)* 4.7 (0.4)
Mixed cells (×109/l) 0.6 (0.1) 0.7 (0.1) 0.7 (0.1) 0.9 (0.1)* † 0.7 (0.1) 0.8 (0.1)* 0.7 (0.1)* 0.7 (0.1)
Platelets (×109/l) 220 (11) 220 (12) 230 (12) 240 (12)** 230 (11)* 240 (11)* 230 (11)* 230 (11)
2612 Eur J Appl Physiol (2014) 114:2607–2616
1 3
of the leucocytosis. In this study, the increase in total WBC
was significantly higher after HYP than MAX. Risoy et al.
(2003) demonstrated that white blood cells infiltrate into
exercised muscles and that might be related to impaired
recovery. In the present study, neutrophil concentrations
peaked immediately after HYP, whereas in MAX the acute
response was slower and became significant 15 min after
the end of the exercise session. In accordance with previous
study (Mayhew et al. 2005), the peak values might have
been observed later than our 30 min time point.
Previous studies (Suzuki et al. 1999; Peake et al. 2005b;
Paulsen and Peake 2013) have reported that post-exercise
myoglobin concentrations correlate with neutrophil counts
especially in the delayed phase of leukocytosis. In the pre-
sent study, during the recovery window of 30 min, a sig-
nificant correlation was observed only in HYP, even though
both MAX and HYP induced similar acute increase in
serum myoglobin concentrations. To avoid repeated bout
effect on inflammatory response that has been shown in
muscle damaging, especially eccentric exercise bouts, the
participants took part into familiarization session in which
1RM was measured and counter-balanced design was used
(McHugh 2003). The main limitation in the present study
was the narrow recovery window which might have hidden
the peak myoglobin levels. On the other hand significant
increase in myoglobin was not observed between POST15
and POST30. The present study observed significant
increases in lymphocytes only after HYP. Interestingly, a
slight, but significant decrease in lymphocyte counts was
observed in MAX at 15 and 30 min. This might indicate
that a high-intensity maximal resistance exercise bout
might induce lymphopenia, which has been associated with
so called “open window theory” of higher risk for infec-
tions after exercise (Nieman and Pedersen 1999). Again,
the last recovery sample was collected 30 min after the
exercise bouts and lymphopenia might be observed later
than that in HYP. We cannot ignore the fact that a decrease
in lymphocytes below pre-exercise values might have been
also observed in HYP, but due to our recovery window and
the different durations of the exercise protocols, we were
unable to observe it. Shear stress and hormonal signals
have been suggested to induce the release of WBC from
marginated pool (Freidenreich and Volek 2012). Elevations
in cortisol are thought to lead to reductions in circulating
lymphocyte counts during post-exercise recovery (Shinkai
et al. 1996). We did not observe a significant correlation
between cortisol response and increase in WBC in HYP but
interestingly, cortisol levels measured immediately after
exercise correlated negatively with WBC in MAX. The
acute WBC response after exercise is most likely a result
of several mechanisms, especially in HYP, which might
blunt the effects of cortisol. Whereas in MAX, even though
the cortisol concentrations decreased, they correlated
negatively with WBC and lymphocyte counts. Miles et al.
(2003) observed an association between lactate accumula-
tion and increased number of lymphocytes after squat exer-
cise. In the present study, as expected, peak lactate levels
Fig. 1 The effect of exercise bouts on serum IL-6, IL-1ra and MCP-
1. MAX maximal, HYP hypertrophic. *significant difference to pre
value, #significant difference between the exercises. (mean ± SEM
*p < 0.05, **p < 0.01, #p < 0.05)
2613Eur J Appl Physiol (2014) 114:2607–2616
1 3
correlated positively with lymphocyte counts after exercise
bout in HYP whereas a negative correlation between lym-
phocyte counts and lactate concentrations was observed at
POST0 and POST15.
Cytokines
The results of this study demonstrated that IL-6 increased
progressively after both HYP and MAX. Contracting skel-
etal muscle synthesizes and releases IL-6 into the inter-
stitium as well as into systemic circulation during exercise.
Local factors seem to be necessary in IL-6 release and syn-
thesis from contracting muscle but systemic factors may
modulate the response. Muscle contraction has been linked
to IL-6 release in several mechanisms, including change
in calcium homeostasis, impaired glucose availability
and formation of reactive oxygen species (Fischer 2006).
HYP lead to a significantly higher blood lactate concen-
tration, which demonstrates the higher metabolic demand.
Whereas, mechanical stress, due to the higher loads, was
expected to be higher in MAX. It is important to note,
however, that the increase in IL-6 levels was relatively
modest, which is in-line with previous studies (Izquierdo
et al. 2009). Izquierdo et al. (2009) reported significant
increases in IL-6 45 min after 5 × 10RM leg press exercise
in untrained men while IL-1ra increase in a statistically sig-
nificant manner only immediately after a training period of
7 weeks. Secreted IL-6 has shown to have an anti-inflam-
matory effect as it stimulates IL-10 and IL-1ra (Steensberg
et al. 2003). Interestingly, even if the exercise bouts were
very different significant differences in IL-6 response were
not observed, we observed a significant increase in IL-1ra
after HYP, but not after MAX, which might indicate that
higher stimulation of the cardiovascular system might be
needed to elicit a response in IL-1ra. In addition, there
was a trend for MCP-1 concentration to decrease below
pre-exercise levels in both loadings during recovery, but
this reached statistical significance only in HYP. MCP-1
has been reported to increase immediately after muscle
damaging exercise (Peake 2005a; Crystal 2013); how-
ever, the immediate increase in MCP-1 was not observed.
MCP-1 and its receptors are required for successful muscle
regeneration, but at the same time MCP-1 has a negative
role in chronic diseases that include low-grade inflamma-
tion (Paulsen et al. 2012; Kim et al. 2006). In addition to
acute increase in IL-1ra in the present study, the significant
decrease in MCP-1 after the HYP could be considered as
an acute anti-inflammatory response to HYP.
Fig. 2 The effect of exercise bouts on serum adipsin, adiponectin, leptin and resistin. MAX maximal, HYP hypertrophic. *significant difference
to pre value. There were no significant differences between loadings. (mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001)
2614 Eur J Appl Physiol (2014) 114:2607–2616
1 3
Adipocytokines
Altered adipocytokine levels (especially low adiponectin)
have been proposed to be a link between obesity and dia-
betes (Kanaya et al. 2004). In the present study, in-line with
previous reports (Ronti et al. 2006), resting adiponectin
levels negatively correlated with body fat. Bouassida et al.
(2008) suggested that to induce significant acute decreases
in leptin, a background in training might be needed and
thus significant adipocytokine response would be observed
only when performed by trained athletes. In the present
study, no significant changes in leptin or adiponectin levels
were observed. In contrast, resistin and adipsin decreased
significantly immediately after both resistance exercise
bouts. (Varady et al. 2010) observed beneficial adipocy-
tokine modulation after resistance exercise only in habitual
weight-trainers, but the present study indicates that HYP,
as well as MAX, may have a beneficial effect on adipo-
cytokine profile also in people who do not have weight
training background when the exercise is performed with
maximum effort. As in the study of (Varady et al. 2010),
the results from this study might only be limited to heavy
resistance exercise bout and for young lean men, and more
research is needed in other populations.
Fig. 3 Significant correlations were observed between fat mass and
adiponectin (r = −0.643, p < 0.05, (a), in HYP, between myoglobin
and IL-1ra at POST0 (r = 0.701, p < 0.05, (b), in MAX, between the
acute increase in WBC and the acute increase in cortisol at POST0
(r = −0.622, p < 0.05, (c). In addition, the peak lactate (POST15)
correlated negatively with lymphocytes in HYP (r = −0.874,
p < 0.05, (d) and in MAX (r = −0.697, p < 0.05, (e)
2615Eur J Appl Physiol (2014) 114:2607–2616
1 3
Exercise protocols
Many types of resistance exercise sessions can be effec-
tively used to improve muscular fitness. The ACSM posi-
tion stand on Progression Models in Resistance Training
for Healthy Adults (2009) recommends loads correspond-
ing to a repetition range 8–12RM to be used for novice
training and 1–12RM in a periodized fashion for intermedi-
ate (individuals with approximately 6 months of consistent
resistance training experience) to advanced training. The
number of sets per exercise recommended is initially one to
three for novice individuals and for progression into inter-
mediate to advanced status it is recommended to use mul-
tiple sets with systematic variation of volume and intensity
over time (American College of Sports Medicine 2009). To
ensure optimal health and fitness gains, resistance training
should be undertaken with proper preparation, guidance
and surveillance (Williams et al. 2007). The exercise pro-
tocols in this study were heavy, which must be taken into
account when evaluating the present results. Studies with a
lower number of sets (Buford et al. 2009) and smaller mus-
cle groups (Uchida et al. 2009) did not observe significant
immediate response in circulating cytokines after a resist-
ance exercise bout. The reported cytokine responses to
resistance exercise vary considerably; however, this study
suggests that heavy exercise protocols may be needed
to elicit a significant change in inflammation markers in
healthy subjects.
Conclusion
The findings of this study suggest that a resistance exercise
bout, in general, produces changes in WBC, cytokines and
adipocytokines. Due to the greater metabolic demand and
stress with shorter rest periods, the WBC and concentra-
tions of cortisol and cytokines increased to a greater extent
after HYP than after MAX. There was notable individual
variability in the cytokine responses, especially in MCP-
1, whereas WBC and adipocytokine responses were more
coherent. Efficient recovery, especially from resistance
exercise, requires a well-coordinated and controlled inflam-
matory response, which includes a rise in pro-inflamma-
tory, as well as anti-inflammatory cytokines. One heavy
resistance exercise bout could lead to an anti-inflammatory
environment, which might reduce inflammation markers
that are related to low-grade inflammation while repeated
in regular training (Gleeson et al. 2011). Future well-con-
trolled studies are needed to determine the mechanisms that
regulate the observed responses and the long-term effects
of repeated heavy resistance exercise bouts on cytokine
and, especially, adipocytokine profiles.
Acknowledgments This project was partly funded by Juho Vainion
Foundation, Ellen and Artturi Nyyssönen Foundation and the Depart-
ment of Biology of Physical Activity, University of Jyväskylä, and
the competitive research funding of Pirkanmaa Hospital District. The
authors would like to thank the subjects and research assistants.
Conflict of interest The authors report no conflicts of interest.
The authors alone are responsible for the content and writing of the
manuscript.
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