Luminescence 2007; 22: 20– 28

20
Published ORIGINAL
online 26 JulyRESEARCH
2006 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/bio.922 N. Mochida et al.
ORIGINAL RESEARCH

The main neutrophil and neutrophil-related functions

may compensate for each other following exercise—
a finding from training in university judoists

Noriko Mochida,1,2 Takashi Umeda,1 Yousuke Yamamoto,1,2 Masaru Tanabe,2 Arata Kojima,1,2
Kazuo Sugawara1 and Shigeyuki Nakaji1*
1
Department of Social Medicine, Hirosaki University School of Medicine, Zaifu-cho 5, Hirosaki, 036-8562 Aomori, Japan
2
Department of Physical Education, Nippon Sport Science University, 7-1-1 Fukasawa, Setagaya, Tokyo 158-8508, Japan

Received 3 October 2005; revised 4 April 2006; accepted 20 April 2006

ABSTRACT: In order to clarify the relationship between exercise and neutrophil function, we measured three major neutrophil
and neutrophil-related functions, viz. the reactive oxygen species (ROS) production capability and phagocytic activity (PA) of
neutrophils and serum opsonic activity (SOA), simultaneously before and after a unified loading exercise under three different
sets of conditions. Thirteen female collegiate judoists were examined with a unified exercise loading (2 h) immediately before and
after a 64 day training period. Immediately thereafter, the athletes took part in a 6 day intensified training camp, following which
the same exercise loading was repeated. Responses from circulating neutrophils were estimated by comparing the two sets of
values obtained before and after the two instances of exercise loading. The parameters assessed included neutrophil count, SOA,
PA and ROS production capability. ROS production increased after the exercise loading performed immediately before and after
the 64 day training period just before the camp, (p < 0.01) but decreased following the exercise loading performed after the camp
(p < 0.05). This suggested depressed bacteriocidal capability of the circulating neutrophils. PA decreased after the exercise load-
ing sessions imposed prior to and after the 64 day training period (p < 0.01) but did not change in the loading session after the
camp. No changes were seen in SOA produced with the loading exercise either before the 64 day exercise period or before the
camp, but increased significantly following the post-camp session (p < 0.05). In conclusion, athletic training-induced changes in
immune functional activities of neutrophils, such as ROS production and PA, and neutrophil-related factors, such as SOA, may
compensate for each other to maintain the overall integrity of the neutrophil immune function. Copyright © 2006 John Wiley &
Sons, Ltd.

KEYWORDS: reactive oxygen species; judo; female; serum opsonic activity; neutrophil phagocytic activity

INTRODUCTION oxygen species (ROS) (1). Serum opsonic activity

For many competitive sports, a special training session
(the so-called ‘camp’) is held to reinforce physical
strength and skill, where, in general, the physical load
level is settled at a very much higher intensity and/or a
longer duration compared with those during ordinary
quotidian training. The potential effects of exercise or
training on the immune system have recently attracted
attention. There has been evidence that exercise causes
changes in the distribution and function of immune
factors, either cellular or humoral. Neutrophils represent
one of the cellular factors and play an important role
in the first line of defence against foreign substances,
including microorganisms. Neutrophils engulf micro-
organisms (phagocytic activity, PA) and produce reactive
*Correspondence to: S. Nakaji, Department of Social Medicine,
Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki,
036-8562 Japan.
E-mail: nakaji@cc.hirosaki-u.ac.jp
Contract/grant sponsor: Ministry of Education, Science and Culture,
Japan; Contract/grant number: 11470092.
(SOA) contributes to this microbicidal activity through
opsonization of microorganisms, i.e. an acceleration
of adhesion of neutrophils to opsonized substances via
IgG, C3 and others (1). ROS from neutrophils can
destroy invading microorganisms (2, 3), but at the same
time excessive levels of ROS can damage normal body
tissues and organs (4, 5).
Prolonged and high-intensity training has been
reported to change the ROS production capability of
neutrophils (5–8). This function and related processes,
however, have attracted some controversy. ROS produc-
tion has been reported to increase (5, 7) or decrease
after acute exercise (5, 8). PA significantly decreased
after intense exercise (9) or, in other reports, increased
or did not change with exercise (4, 5, 10, 11). SOA in
long-distance runners after a 1 month camp did not
change (12) or increased after a 30 km run (13). The
reason for these contradictory findings is unknown,
although it has been suggested that it may be due to vari-
ations in the study designs, in which the degree of physi-
cal load and its duration applied to each athlete, their

Copyright © 2006 John Wiley & Sons, Ltd. Luminescence 2007; 22: 20–28
DOI: 10.1002/bio
Exercise and neutrophil function
current physical condition (fitness and fatigue) and
specimen sampling conditions (atmospheric tempera-
ture, timing of sampling, especially the lapse from the
completion of physical load) seem crucial.
In this study, in order to clarify the contradictory
findings in previous studies of the relationship between
exercise and neutrophil function, we measured the
major neutrophil functions, viz. their reactive oxygen
species (ROS) production capability and phagocytic
activity (PA), and serum opsonic activity (SOA) as a
neutrophil-related function, simultaneously, before and
after unified exercise loading under three different con-
ditions: before and after a normal 64 day training period
and after an intense 6 day training session (training
camp) held immediately after the 64 day period of
normal training.
SUBJECTS
Thirteen female collegiate judoists were enrolled in this
study. One student competed in the under-48 kg class,
four in the under-52 kg class, two in the under-57 kg
class, three in the under-63 kg class, two in the under-
78 kg class and one in the over-78 kg class. The average
age, height and weight of the athletes at the commence-
ment of the this investigation were 19.6 ± 1.0 years,
160.3 ± 6.7 cm and 62.1 ± 6.7 kg, respectively.
METHODS
Study protocol
All students completed a normal 64 day quotidian train-
ing session on 8 June–10 August 2004; 2 days after this
they attended a training camp on 12–17 August. Meas-
urement and blood sampling were performed on the day
before the beginning (7 June) and the day after the end
(11 August) of the 64 day normal training period, and
the day after the end (18 August) of the much more
intense training camp.
Early in the morning of each assessment day, the same
standardized exercise regimen (‘exercise loading’) was
imposed on all students under fasting conditions. The
exercise loading was a means to apply the same physi-
cal stressor to the athletes to see responses of circulat-
ing neutrophils to this stress, with the expectation of
differing modulations of neutrophil response compared
between the 64 day normal and 6 day intensive training
programmes.
Blood work-ups were performed before and after
the loading. The mean values of ambient temperature
and humidity were 24.8 ± 0.4°C and 67.4 ± 1.5% on
7 June, 28.2 ± 0.4°C and 70.2 ± 3.5% on 11 August, and
23.7 ± 0.5°C and 85.8 ± 4.8% on 18 August. None of
Copyright © 2006 John Wiley & Sons, Ltd.
ORIGINAL
ORIGINALRESEARCH
RESEARCH 21
the students was under a weight-reduction regimen for
competition.
Approval for the aims and study protocol was ob-
tained from the Ethical Review Committee of Hirosaki
University School of Medicine. Each student was
adequately informed of the aim, methods and potential
risks as well as the right to abstain from participation in
the study. Freely-given informed consent in writing was
obtained from all the students.
Physical load
Loading exercise. The formulated exercise loading
consisted of: warming-up for 15 min; ‘uchikomi’ (repeti-
tion of an art skill in the order, throw down, push down
and hook down) for 20 min; ‘randori’ (actual match
practice) for 70 min; and cooling-down for 15 min. Meas-
urement and blood sample collection were performed
prior to and after this physical load.
Normal daily training. Ordinary training during the
64 day period consisted of judo skills practice for 2.5 h
and running or weight training for 1 h each day. The
students repeated these for 6 days and then took a 1 day
rest (Table 1). This practice pattern was repeated nine
times during the 64 day training period.
Intensifying training. The intensifying training during
the camp consisted of judo skills practice for 3.5 h in the
morning and running (a combination of long-distance
and short sprints) for 2 h followed by weight training for
1 h in the afternoon. The students in the camp worked
for a total of 6.5 h each day, which was nearly double
that of an ordinary training day.
Measurements and blood work-ups
Body composition. The anthropometric parameters
measured were body weight (BW), relative body fat
(% fat) and fat-free mass (FFM), measured by the
impedance method (TBF-110, TANITA, Tokyo,
Japan).
Blood analysis. Blood samples were taken from the
forearm vein before and after the exercise loading
session at each of the three assessment points. The
serum was separated from the blood by centrifugation
for 10 min at 3000 r.p.m. and kept frozen at −30°C until
analysis. White blood cell (WBC) and neurophil counts,
haematocrit (Hct) and haemoglobin (Hb) were deter-
mined using a blood cell autoanalyser (Micro Biff-II,
Coulter Co. Ltd, CA, USA).
The following were assessed: levels of serum enzymes
related to physical load or stress; immunoglobulins and
complements; ROS production; PA; SOA; CD11b (com-
plement receptor type 3: CR3) and CD16 (Fc gamma
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22 ORIGINAL RESEARCH
Table 1. Ordinary training and 1 week exercise programme (1 unit)
6:30 a.m.–7:30 a.m.
Monday Training A
Tuesday Training B
Wednesday Training C
Thursday Training A
Friday Training B
Saturday Training C
Sunday Rest
Training A, interval training consisted of sprint running (800 m × 1, 400 m × 3, 200 m × 3, 100 m × 4) and
jogging.
Training B, weight training.
Training C, distance run for 30 min and short sprint running (repeated 30–50 m sprint running during
training time).
Training D, judo skills practice.
Rest, resting or attending lectures.
receptor type 3: Fcγ R3) expressed on the surface of
neutrophils:
1. Serum enzymes and immunoglobulins. Serum
creatine kinase (CK), lactate dehydrogenase (LDH),
aspartate aminotransferase (ASAT) and alanine
aminotransferase (ALAT) were measured using a
biochemical assay kit (OLYMPUS AU-5232, Tokyo,
Japan). Immunoglobulins (IgG, IgA and IgM) and
complements (C3 and C4) were assayed using a
turbidimetric immunoassay kit (Nittobo Medical Co.
Ltd, Tokyo, Japan).
2. ROS production capability, PA and CD11b and CD16
expression. These neutrophil activities were deter-
mined using a FACScan system (Becton Dickinson,
San Jose, CA) using two-colour flow cytometry.
Hydroethidine (HE: 44.4 µmol/L; Polysience Inc.,
Warrington, PA) was used as the indicator for
oxidative burst activity (ROS production capability),
and opsonized zymosan (OZ) particles labelled with
fluorescein isothiocyanate (FITC, Sigma) for PA (14,
15). Zymozan was purchased from Sigma Chemical
Co. (St. Louis, MO, USA).
a) Preparation for neutrophil oxidative burst and PA.
Whole blood (100 µL) was mixed with 22 µL HE
(8 µmol/L), and incubated at room temperature.
After the addition of 25 µL FITC-labelled
opsonized zymosan [FITC–OZ: final concentra-
tion (f.c.) 5 mg/mL], the sample was incubated
at room temperature. The same amount of
whole blood labelled only with HE was prepared
to measure the basal oxidative burst activity.
Extracellular fluorescence was quenched by add-
ing 30 µL Trypan blue (0.25 mg/mL, pH 4.5) just
before the assay to differentiate between attached
and ingested FITC–OZ in the neutrophils (14, 15).
b) Preparation for CD11b and CD16 expression on
the neutrophil surface. Monoclonal antibodies to
CD 11b and CD16 (IMMUNOTECH) were used
Copyright © 2006 John Wiley & Sons, Ltd.
N. Mochida et al.
9:00 a.m.–11:30 a.m. 5:30 p.m.–8:00 p.m.
Rest Training D
Rest Training D
Rest Training D
Rest Training D
Rest Training D
Training D Rest
Rest Rest
to measure the expression on neutrophils. A
100 µL sample of heparinized whole blood was
mixed with the monoclonal antibodies and incu-
bated at room temperature.
3. Measurement of SOA.
a) Chemiluminogenic probe. The chemiluminogenic
probe, lucigenin (Lg) was prepared by dissolving
bis-N-methylacridinium nitrate (Sigma, USA) in
HBSS to give a final concentration of 0.5 mmol/L
(pH 7.4).
b) Measurement of chemiluminescence. Zymosan A
suspension (5 mg/mL) was used to opsonize each
serum sample at a final concentration of 20%.
OZ was washed twice with HBSS to eliminate
any remaining serum. Standard neutrophils were
obtained from the peripheral blood of a healthy
volunteer using Histopaque 1077 and 1119 (Sigma,
St. Louis, CA, USA). Neutrophil solution (50 mL)
was added to each opsonized zymosan sample
together with the Lg, and the level of lumines-
cence was measured on an Auto Luminescence
Analyzer, Alfa system (Tokken, Funabashi,
Japan) (16). The area under the curve (AUC) of
the chemiluminescence response was used in this
study as described previously (17, 18.)
Statistical analysis
Data were expressed as means ± standard deviations
(SD). Differences between means were examined by a
two-way analysis of variance (ANOVA) with post hoc
tests. The relationship between change rates among PA
per cell, ROS production per cell, the AUC of SOA,
CD11b/CD16 expression per cell, immunoglobulins and
complements were studied using Spearman’s rank order
correlation test. Differences were considered statistically
significant at p < 0.05.
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Exercise and neutrophil function ORIGINAL
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RESEARCH 23

Table 2. Serum myogenic enzymes

Before the 64-day

training period Before the camp After the camp
ASAT (IU/l)
Pre 20.2 ± 3.6 18.4 ± 4.7 21.9 ± 4.7
Post 23.4 ± 3.5** 19.9 ± 4.8** 25.6 ± 5.6**
Change-ratio (%) 16.1 ± 6.3 8.6 ± 5.3†† 17.0 ± 5.6
ALAT (IU/l)
Pre 15.0 ± 3.5 17.5 ± 11.8 17.1 ± 6.4
Post 16.5 ± 3.5** 18.1 ± 12.9* 19.4 ± 7.2**
% Change 10.8 ± 8.6 2.1 ± 8.1† 13.4 ± 5.9‡‡
CK (IU/l)
Pre 225.0 ± 70.8 133.1 ± 50.2 300.5 ± 94.2
Post 273.4 ± 69.1** 168.6 ± 54.9** 365.7 ± 112.9**
% Change 23.4 ± 7.4 28.6 ± 11.5 21.9 ± 6.8
LDH (IU/l)
Pre 221.3 ± 37.4 188.1 ± 37.5 237.6 ± 25.5
Post 266.4 ± 40.5** 217.1 ± 24.5* 276.1 ± 24.4**
% Change 20.9 ± 8.1 17.3 ± 12.5 16.5 ± 6.1

*Significant at p < 0.05 and **at p < 0.01 between pre- and post-values.
†
Significant at p < 0.05 and ††at p < 0.01 when compared with the rate of change before the normal
64-day training period.
‡
Significant at p < 0.05 and ‡‡at p < 0.01 when compared with the rate of change before the camp.

RESULTS
Body composition
Body weight significantly decreased after the exercise
loading (p < 0.01 for all) on all of the three assessment
days. Body height, %fat and FFM did not change
throughout the investigation.
Serum enzymes
ASAT, LDH and CK significantly increased (p <
0.05−0.01) after the exercise loading on all of the three
assessment days (Table 2).
Leukocyte and neutrophil counts
Leukocyte counts significantly increased (p < 0.01 for all)
after the loading exercise at the beginning of the 64 day
normal training period training and after the 6 day train-
ing camp. Neutrophil counts significantly increased after
the exercise loading on all assessment days (p < 0.01 for
all). A significant increase in the neutrophil:leukocyte
ratio (p < 0.01 for all) was seen both before and after the
64 day training period (Table 3).
Immunoglobulins and complements
IgG, IgA and IgM significantly increased after the exer-
cise loading on all of the three assessment days (p < 0.01
for all) (Table 4), as did C3 and C4 (p < 0.05−0.01)
(Table 4).
ROS production capability, PA and expression
of CDs
ROS production significantly increased after the exercise
loading both before and after the 64 day training session
(p < 0.01), whereas it significantly decreased (p < 0.05)
after the camp (Table 5). PA significantly decreased
(p < 0.01) after the exercise loading, both before and
after the 64 day training session, and remained
unchanged after the camp (Table 5). No apparent
change was seen in the expressions of CD11b and CD16
(Table 6).
SOA
AUC of the light emission showed no change before and
after the 64 day training session, but significantly
increased (p < 0.05) after the camp (Table 7).
Correlation of major factors of neutrophil
function
Table 8 shows Spearman’s correlation coefficients
among the change rates in ROS production, PA, the
expression of CD11b or CD16, and SOA. Negative
correlations were seen between ROS production and
PA (r = −0.322; p < 0.05) and ROS production and SOA
(r = −0.368; p < 0.05) (Fig. 1). A significant correlation
was also seen in PA vs. CD11b expression (r = −0.302;
p < 0.05). On the other hand, the rates of change in
immunoglobulins and complements had no correlation
with those in SOA.

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DOI: 10.1002/bio
24 ORIGINAL RESEARCH N. Mochida et al.

Table 3. Leukocyte and neutrophil counts

Before the 64-day

training period Before the camp After the camp
Leukocyte counts (/µL)
Pre 5264 ± 1299 6193 ± 1383 5321 ± 892
Post 6407 ± 1431** 6957 ± 1913 6593 ± 1375**
% Change 24.4 ± 21.8 12.5 ± 20.7 24.8 ± 21.9
Neutrophil counts (/µL)
Pre 2450 ± 978 3248 ± 1107 3169 ± 726
Post 3684 ± 1100** 4645 ± 1546** 4126 ± 826**
% Change 66.7 ± 67.8 51.0 ± 50.1 34.4 ± 34.6†
Neutrophil counts/leukocyte counts × 100 (%)
Pre 45.3 ± 10.9 51.5 ± 11.0 59.3 ± 6.9
Post 57.5 ± 10.7** 65.7 ± 7.4** 63.0 ± 7.3
% Change 31.7 ± 35.5 32.2 ± 28.4 7.1 ± 14.3†‡

*Significant at p < 0.05 and **at p < 0.01 between pre- and post-values.
†
Significant at p < 0.05 and ††at p < 0.01 when compared with the rate of change before the 64-day
training period.
‡
Significant at p < 0.05 when compared with the rate of change before the camp.

Table 4. Serum immunoglobulins and complements

Before the 64-day

training period Before the camp After the camp
IgG (mg/dL)
Pre 1170 ± 261 1109 ± 235 1066 ± 241
Post 1239 ± 270** 1162 ± 265** 1149 ± 270**
% Change 6.0 ± 3.8 4.4 ± 5.6 7.6 ± 4.5
IgA (mg/dL)
Pre 220.4 ± 110.0 206.7 ± 90.2 199.4 ± 101.1
Post 230.1 ± 119.9** 212.5 ± 93.8** 211.1 ± 103.4**
% Change 3.7 ± 3.7 2.6 ± 3.8 6.7 ± 3.7
IgM (mg/dL)
Pre 151.1 ± 57.0 146.0 ± 55.7 134.9 ± 47.5
Post 157.2 ± 61.1** 153.6 ± 61.8** 143.1 ± 53.0**
% Change 3.8 ± 4.0 4.3 ± 4.7 5.5 ± 3.4
C3 (mg/dL)
Pre 92.9 ± 13.4 98.2 ± 15.3 92.1 ± 16.3
Post 95.2 ± 14.7** 102.8 ± 14.5* 97.0 ± 17.5**
% Change 2.4 ± 4.1 4.9 ± 4.4 5.2 ± 3.5
C4 (mg/dL)
Pre 19.6 ± 3.8 21.7 ± 4.5 21.8 ± 4.0
Post 19.9 ± 3.9* 22.8 ± 4.5** 22.7 ± 4.3*
% Change 1.9 ± 4.5 5.2 ± 3.5 4.2 ± 4.0

*Significant at p < 0.05 and **at p < 0.01 between pre- and post-values.

DISCUSSION in these enzyme levels may be effective markers for

Exercise has been reported to elevate the levels of such
serum enzymes as CK, ASAT, ASLT and LDH through
the possible mechanisms of muscular inflammation,
increased permeability of muscle cell membranes (19,
20), anoxic or hypoxic damage to muscles (21, 22) or
production of toxic free radicals (23). Therefore, changes
monitoring overtraining in athletes (19). Evans et al.
reported that CK levels were raised more remarkably
in non-exercising subjects than in habitually exercising
subjects under the same exercise load, and that habitual
exercise might increase the capability of muscular
function and tissue structure to alloy the performance of
more intense exercise without any exercise-related

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Exercise and neutrophil function ORIGINAL
ORIGINALRESEARCH
RESEARCH 25

Table 5. ROS production and PA

Before the 64-day

training period Before the camp After the camp
ROS production per cell (FI)
Pre 269.2 ± 30.1 270.2 ± 31.0 318.6 ± 26.3
Post 299.3 ± 32.0** 295.8 ± 33.7** 278.8 ± 75.0*
% Change 11.7 ± 11.6 9.8 ± 9.2 −13.1 ± 20.7††‡‡
PA per cell (FI)
Pre 565.4 ± 109.8 524.8 ± 57.1 462.3 ± 44.7
Post 437.3 ± 27.3** 440.1 ± 66.2** 455.1 ± 75.5
% Change −20.8 ± 11.4 −16.0 ± 9.2 −0.8 ± 19.7††‡‡

FI, fluorescence intensity.

*Significant at p < 0.05 and **at p < 0.01 between pre- and post-values.
††
Significant at p < 0.01 when compared with the rate of change before the 64-day training period.
‡‡
Significant at p < 0.01 when compared with the rate of change before the camp.

Table 6. CD11b and CD16 expression

Before the 64-day

training period Before the camp After the camp
CD11b per cell (FI)
Pre 125.9 ± 17.3 128.0 ± 25.0 137.0 ± 19.4
Post 136.2 ± 32.8 126.2 ± 23.5 140.3 ± 30.5
% Change 7.4 ± 15.6 −0.5 ± 14.8 4.5 ± 31.7
Positive rate of CD11b (%)
Pre 93.2 ± 3.2 95.1 ± 1.4 96.3 ± 1.2
Post 95.3 ± 1.3 96.2 ± 1.5 96.4 ± 1.7
% Change 2.1 ± 2.8 1.1 ± 0.9 0.1 ± 1.5
CD16 per cell (FI)
Pre 1564 ± 494 1434 ± 426 1484 ± 504
Post 1482 ± 448 1385 ± 491 1407 ± 441
% Change −4.3 ± 12.0 −4.5 ± 8.1 −3.8 ± 14.5
Positive rate of CD16 (%)
Pre 82.5 ± 10.1 84.0 ± 10.5 88.5 ± 5.8
Post 89.8 ± 8.0* 90.0 ± 8.4 91.8 ± 5.3
% Change 7.4 ± 6.6 5.9 ± 4.0 3.2 ± 3.2

Table 7. SOA

Before the 64-day

training period Before the camp After the camp
AUC
Pre 2848 ± 535 570 ± 77 2176 ± 226
Post 2734 ± 904 590 ± 78† 2408 ± 305*†
% Change −13.9 ± 27.6 4.7 ± 15.7 11.8 ± 18.5

AUC, area under the curve for 45 min.

*Significant at p < 0.05 between pre- and post-values.
†
Significant at p < 0.05 when compared with the rate of change before the 64 day training period.

Table 8. Spearman’s correlation coefficients between two parameters

ROS production AUC of CD11b CD16

PA per cell per cell SOA expression/cell expression/cell
PA per cell – – – – –
ROS production per cell −0.322* – – – –
AUC of SOA ns −0.368* – – –
CD11b expression per cell −0.302* ns ns – –
CD16 expression per cell ns ns ns ns –

*Significant at p < 0.05.

ns, not significant.

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DOI: 10.1002/bio
26 ORIGINAL RESEARCH
Figure 1. Correlation between the rates of change in ROS production, PA and SOA.
muscle injury (24). Koutedakis et al. also reported that
intense high-quality physical training in Olympic rowers
failed to produce any significant change in enzyme
levels, and speculated that an inevitable adaptation had
occurred in such elite competitors, i.e. a blunted incre-
mental response to exercise (20). The present study con-
firmed the previous findings (19, 20, 22, 23) that exercise
increased the levels of serum CK, ASAT, ASLT and
LDH. Although the athletes in the present study were
well-trained national top-class judoists, the increase
in the level of each serum enzyme after the loading
exercise was statistically significant on the three differ-
ent occasions of assessment. The 2 h exercise loading
was specifically formulated for the assessments, but was
apparently sufficiently intense to damage muscle tissue.
The increase in the number of circulating neutrophils
with exercise has been well established, and the increase
is in an intensity-dependent fashion (25). This increase
may be as a result of stress-related release of growth
hormone, adrenalin or noradrenalin (2) or a part of the
inflammatory response via cytokines to exercise-induced
tissue damage (26). In the present study, the increases
in neutrophil counts were statistically well correlated
with the elevation of the enzyme levels. However, it was
beyond the scope of this study to ascertain whether the
Copyright © 2006 John Wiley & Sons, Ltd.
N. Mochida et al.
increased number of neutrophils was a cause of muscle
damage through enhanced ROS production or a result
of compromised muscle cells being engulfed.
The immune system is extremely sensitive to stress,
whether physical or psychological (27). Therefore the
immune system can be used as a sensor of stress for
monitoring the impact of exercise and training on health.
In this study, the immune function of neutrophils, a
cellular component of the immune system, was assessed
by measuring ROS and PA in addition to SOA as a
neutrophil-related function.
ROS production significantly increased after the exer-
cise loading both at the beginning and end of the 64 day
normal training session (p < 0.01), whereas it decreased
(p < 0.05) after the camp. As already mentioned,
enhanced ROS production works beneficially for bacte-
ricidal action (2, 3) but can work adversely by causing
injury to normal tissue at higher concentrations (4, 5):
however, which of these two scenarios was dominant
at these two stages of the training protocol remains
unelucidated. PA significantly decreased after the exer-
cise loading both at the beginning and end of the 64 day
training session (p < 0.01) but it did not change after the
camp, while SOA showed no change following exercise
loading before and after the 64 day training session, but
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Exercise and neutrophil function
Figure 2. Compensatory relationships among ROS, PA and SOA. ROS production changed in the reverse
direction to SOA or PA. These functions may compensate for each other to maintain the overall integrity of
neutrophil function (immune homeostasis).
showed a significant increase after the post-camp exer-
cise loading (p < 0.05). The relationships among the
change rates of ROS, SOA and PA are illustrated in
Fig. 2, from which it can be seen that ROS production
moved in the reverse direction to SOA or PA. In other
words, the ROS rate of change (% change) was inversely
correlated with those of SOA (r = −0.322; p < 0.05) and
PA (r = −0.368; p < 0.05). This finding could be inter-
preted speculatively to show that these three functions
may compensate for each other to maintain the overall
integrity of neutrophil function, i.e. a part of immune
homeostasis.
On the other hand, the rates of change in immuno-
globulins and complements, even though they are
known to affect SOA, had no correlation with the SOA
change rate. The rates of change in the CD receptors on
neutrophils, through which PA is efficiently executed,
had no correlation with the PA change rate. Therefore,
factors other than immunoglobulins and complements or
CD receptors may be involved in SOA and PA and may
have a larger influence on them.
Acknowledgements
The authors would like to thank Dr Daisuke Chinda, Dr
Ippei Takahashi and Dr Takao Oyama, Department
of Social Medicine, Hirosaki University School of
Medicine, for their suggestions while preparing this
Copyright © 2006 John Wiley & Sons, Ltd.
ORIGINAL
ORIGINALRESEARCH
RESEARCH 27
manuscript. This work was supported by a grant-in-aid
for scientific research from the Ministry of Education,
Science and Culture of Japan (No. 11470092).
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