Effects of concurrent training on inflammatory markers and expression of CD4, CD8, and HLA-DR in...

Available online at www.sciencedirect.com

+ MODEL

ScienceDirect

Journal of Exercise Science & Fitness xx (2014) 1e7www.elsevier.com/locate/jesf

Original article

Effects of concurrent training on inflammatory markers and expression ofCD4, CD8, and HLA-DR in overweight and obese adults

Alana Colato a, Fabiana Abreu a, Niara Medeiros a, Leandro Lemos a, Gilson Dorneles a,Thiago Ramis a, Priscila Vianna b, Jos�e Artur Chies b, Alessandra Peres a,c,*

a Centro Universit�ario Metodista do IPA, Laboratory of Immunology and Exercise Physiology, Porto Alegre, Brazilb Universidade Federal do Rio Grande do Sul, Department of Immunogenetics, Porto Alegre, Brazil

c Universidade Federal de Ciencias da Saude de Porto Alegre, Laboratory of Immunology, Porto Alegre, Brazil

Received 10 October 2013; revised 27 March 2014; accepted 3 June 2014

Abstract

The number of people who are overweight or obese is increasing worldwide and the quality of life of these people can be affected by theircondition. Physical training has been studied in obese patients and is correlated with low-grade inflammation and alterations in the immunesystem. This study investigated the effect of concurrent training on anthropometric, inflammatory, and immunological parameters in overweightand obese adults. Fourteen sedentary volunteers (men and women) with a body mass index between 25 kg/m2 and 39.9 kg/m2 from Porto Alegre,Brazil attended a 12-week course of concurrent training. We analyzed: prior to and after training, anthropometric parameters, cytokine serumlevels (interleukin 6, tumor necrosis factor a, interferon g, interleukin 17A and interleukin 10; measured by enzyme-linked immunosorbentassay), high-sensitive C reactive protein (measured by turbidimetry), and the frequency of T lymphocytes and monocytes (CD3þCD4þ,CD3þCD8þ, and HLA-DRþ) in peripheral blood (measured by flow cytometry). The sample consisted of ten women and four men with amean ± SD age of 47.58 ± 3.01 years. After 12 weeks of training we observed a reduction in body weight, body mass index, waist, abdomen,and hip circumferences, the percentage mass of fat, and an increase in the time taken to reach exhaustion (p < 0.05). The participants hadincreased frequencies of CD3þCD4þ and CD3þCD8þ T lymphocytes and a reduction in the frequencies of HLA-DRþ monocytes (p < 0.05).Interestingly, the levels of tumor necrosis factor a and high-sensitive C reactive protein increased (p < 0.05). Our data suggest that concurrenttraining can improve body composition as well as increasing T cell proliferation in overweight and obese patients. However, the progression ofthe exercises can be physiologically stressful to these patients, as demonstrated by the inflammatory markers.Copyright © 2014, The Society of Chinese Scholars on Exercise Physiology and Fitness. Published by Elsevier (Singapore) Pte Ltd. All rightsreserved.

Keywords: Chronic exercise; Cytokines; Inflammation; Monocytes; T lymphocytes

Introduction

Obesity is a chronic and multifactorial disease that isincreasing worldwide and is associated with the increasingincidence of cardiovascular disease, cancer, metabolic

* Corresponding author. Rua Joaquim Pedro Salgado, 165/201 Bairro Rio

Branco, Porto Alegre, RS, Brazil.

E-mail address: [email protected] (A. Peres).

Please cite this article in press as: Colato A, et al., Effects of concurrent trainin

overweight and obese adults, Journal of Exercise Science & Fitness (2014), http:

http://dx.doi.org/10.1016/j.jesf.2014.06.002

1728-869X/Copyright © 2014, The Society of Chinese Scholars on Exercise Physiology and

syndrome, and type 2 diabetes mellitus.1,2 Epidemiologicalstudies indicate a strong link between a high calorie intake/sedentary lifestyle and a dramatic increase in the incidence ofobesity.3 In Brazil, about 50% of the population is overweight,which makes obesity a serious public health problem.3 It iswell known that obesity is associated with a state of chronic,low-grade inflammation, which is involved in the pathogenesisof the related comorbidities. Previous studies have shownincreased levels of inflammatory markers, including inter-leukin 6 (IL-6), tumor necrosis factor a (TNF-a), C reactive

g on inflammatory markers and expression of CD4, CD8, and HLA-DR in

//dx.doi.org/10.1016/j.jesf.2014.06.002

Fitness. Published by Elsevier (Singapore) Pte Ltd. All rights reserved.

mailto:[email protected]

www.sciencedirect.com/science/journal/1728869X




http://www.elsevier.com/locate/jesf


2 A. Colato et al. / Journal of Exercise Science & Fitness xx (2014) 1e7

+ MODEL

protein (CRP), and complement factors, in obese patients.4e6

Moreover, adipose tissue, especially visceral adipose tissue,is an important source of cytokines and patients with excessadiposity show high levels of these inflammatory mediators.7,8

The large release of inflammatory markers in adipose tissuemay also be due to infiltration by macrophages and otherimmune cells, which show increased volumes and activity.9 Inaddition, obesity has also been associated with low levels ofIL-10 and adiponectin, which have anti-inflammatory andanti-atherogenic properties.10

Weight loss and exercise training are associated with adecrease in inflammation and insulin resistance in obesepatients.11e13 However, the relationship between physicalexercise and the anti-inflammatory effects is complex anddepends on gender, the degree of obesity, and the generalhealth of the patient. The physical training may paradoxicallytrigger an acute inflammatory response over a short period, butreduced inflammation is related to a long period of exercise.12

The level of cytokines such as TNF-a, IL-6, and CRP seems tobe favorably modulated by continued physical activity andphysical training in obese young adults, increasing insulinsensitivity.10,14e16 Training programs involving aerobic andstrength exercises of moderate intensity can induce benefits inobese patients and patients with type 2 diabetes, such as im-provements in the action of insulin, endothelial function, andlean body mass, which are associated with improved glycemiccontrol.17e19 However, the type, intensity, and duration ofexercise can influence the inflammatory response in somepopulations.20

As the benefits of physical exercise on inflammatory pa-rameters are still controversial, the aim of this study was toevaluate the effect of 12 weeks of concurrent training on theserum levels of IL-6, TNF-a, interferon g (IFN-g), IL-17A,IL-10, and high-sensitive CRP (hs-CRP), as well as theexpression of CD3þCD4þ, CD3þCD8þ, and HLA-DRþ Tlymphocytes and monocytes in sedentary overweight andobese patients.

Materials and methods

Participants and study design

This study was characterized as a quantitative experi-mental study of 14 volunteers recruited by convenience fromthe city of Porto Alegre, Brazil. The inclusion criteria for thepatients were: (1) sedentary; (2) body mass index (BMI)between 25 kg/m2 and 39.9 kg/m2; (3) older than 18 yearsold; (4) both genders; and (5) a medical certificate approvingthe practice of physical training. We excluded patients whohad some disease hindering the practice of physical exercise,such as cancer, chronic infection, or autoimmune disease.Smokers and patients with a different BMI to that specifiedwere also excluded. All patients participating in this studygave written informed consent. This study was approved bythe Research Ethics Committee at the Centro Universit�arioMetodista do IPA (Porto Alegre, Brazil), under protocolnumber 48/12.



Blood samples and anthropometric measurements

We collected 15 mL of peripheral blood by venepuncture(between 7:00 AM and 10:00 AM) after an overnight fast of 12hours. The blood samples were divided into tubes withoutanticoagulant to obtain the serum and into heparinized tubesfor whole blood (BD Vacutainer heparin tubes). Serum sam-ples were separated by centrifugation for 10 minutes at2500 rpm 1048 g, divided into aliquots, and frozen at �20�Cfor further analysis. The heparinized whole blood wasanalyzed within 3 hours of collection. Forty-eight hours afterthe last concurrent training session, blood samples werecollected again. Body weight and height were determined by asemi-analytical scale (Welmy, Santa B�arbara do Oeste, S~aoPaulo, Brazil) with a capacity for 200 kg and an attachedstadiometer, with accuracies of 0.1 kg and 0.005 cm, respec-tively. Body composition was assessed by the bioelectricalimpedance method (Byodinamics, BIA 310e, Seattle, Wash-ington, USA), following all guidelines pre-test, and consid-ering the values of mass percentage fat and free fat mass (kg).We also measured the waist, abdomen, and hip circumfer-ences. The BMI of each participant was calculated by dividingthe weight (kg) by the height squared (m). All assessmentswere performed by a trained exercise physiologist.

Peak oxygen uptake

Peak oxygen uptake (VO2PEAK) was assessed through an in-cremental test until exhaustion on a motorized treadmill(Inbramed Millennium ATL, Porto Alegre, Brazil). The gaseswere collected using an ergoespirometer (V02000, Med-graphics, St Paul, MN, USA). Heart rate (HR) was monitoredduring the test using a frequency counter (POLAR FT7,Tempere, Finland) and blood pressure was checked with asphygmomanometer (Solidor, Hebei Medicinis and HealthProd., China). Throughout the test, the participants used amask (Medgraphics, St. Paul, Minnesota, USA) connected to agas analyzer, which allowed the collection and storage of datafor each breath (breath by breath) using Aerograph (St. Paul,Minnesota, USA) software. The test was conducted accordingto the modified Bruce21 protocol, which consists of stages ofincrement of intensity by increasing the speed or incline of thetreadmill. After each stage the participant reported theirperception of exertion based on the Borg22 scale so that theintensity of the stage could be reproduced during the period ofphysical training. The test was interrupted in the presence ofany of the following events: (1) HR with values above 10 bpmof the maximum predicted for age; (2) signs and symptoms ofangina; (3) decrease or absence of significant increase insystolic blood pressure with increased intensity of training; (4)respiratory quotient above 1.1; or (5) voluntary exhaustion.The VO2PEAK was defined as the highest value obtained after thetreatment of data by three researchers blinded (or independent)to the study. In addition, the time to exhaustion was recordedas a variable of endurance performance. Verbal encourage-ment was provided to ensure that the maximum values werereached.


//dx.doi.org/10.1016/j.jesf.2014.06.002

3A. Colato et al. / Journal of Exercise Science & Fitness xx (2014) 1e7

+ MODEL

Maximum dynamic strength test

The maximum dynamic strength was measured by testingone maximum repetition (1-RM) for the prescription of thestrength exercises.23 The patients performed a warm-up withdynamic movements, followed by a set of 10 maximum rep-etitions at approximately 50% of the estimated 1-RM. After 3minutes, tests were performed with progressive loads for 1-RM and were determined within five attempts applied withan interval of 3e5 minutes between each trial. The 1-RM testwas performed for the lower body by a squat exercise and forthe upper body by a supine exercise with free weights prior tostarting the training, during Week 6, and at the end of thetraining period (Week 12). The test consisted of obtaining themaximum amount of weight that an individual could bear in acomplete cycle (concentriceeccentric) of exercises in a singlerepetition. Strong verbal encouragement was given throughoutthe test. Prior to starting the training, the patients were sub-mitted to familiarization sessions consisting of two series ofdynamic movements without workload to reduce the effects ofmotor learning.

Concurrent training

Concurrent training is described as the execution, in thesame training session, of both aerobic and strength exercises.The concurrent training used in this study is described in Table1, in addition to the linear periodization adopted based on theguidelines of the American College of Sports Medicine.24 Itwas performed during three sessions per week with a minimuminterval of 48 hours over a 12 week period. The pre-trainingweek was used for familiarization and testing of maximummuscle strength (1-RM). The training sessions were dividedinto three parts: (1) warm-up exercises; (2) concurrent training;and (3) stretching. The duration of each training session was 60minutes. Strength training was based on the method of alter-nating segments with the following exercises: bench press withdumbbells plan; squats; shoulder abduction; plantar flexion;horizontal pulley; and abdominal crunch. During training, weapplied a progression of intensity (50e75% 1-RM) as well as adecreased number of repetitions in all exercises, except forabdominal and plantar flexion. The abdominal and plantarflexion were performed with no load and fixed at 15 repetitions.The aerobic training (oriented walking) had a fixed duration of

Table 1

Description of 12-week concurrent training period.

Week 1e3 4e6 7e9 10e11 12

Strength training

Series 3 3 3 3 3

Repetitions 15 12 10 08 15

Intensity (% 1-RM) 50 60 70 75 50

Time of recovery (s) 60 90 90 90 60

Aerobic training

Volume/time (min) 30 30 30 30 30

Intensity (% VO2PEAK) 50 60 70 75 50

Borg scale 12 13 14 15 12

1-RM ¼ one repetition maximum test; VO2PEAK ¼ peak oxygen uptake.



30 minutes per session with a progressive intensity (50e75%VO2PEAK) based on the Borg22 scale and VO2peak values foundduring the cardiorespiratory test.

High-sensitive C reactive protein and cytokine levels

The serum levels of hs-CRP were evaluated by turbidimetry(C-Reactive Protein Reagent, Beckman Coulter Inc, Brea,California, USA) in an automated Advia 1800-Siemens in-strument (Laborclin, Pinhais, Paran�a, Brazil). The cytokinesIL-6, TNF-a, IL-10, INF-g, and IL-17A were determined byenzyme-linked immunosorbent assay (ELISA) using serumsamples (Mini ELISA Development Kit, 900-M21, PeproTechInc, New Jersey, USA).

Flow cytometry

Immunophenotyping was carried out using a direct immu-nofluorescence technique. Monoclonal antibodies specific forCD3, CD4, CD8, and HLA-DR, as well as the appropriate iso-type controls, were purchased fromBectonDickinson (San Jose,CA, USA). We used a two-color staining method using mono-clonal antibodies labeled with fluorescein isothiocyanate andantibodies labeled with phycoerythrin added to 12 � 75 mmtest-tubes. One hundred microliters of heparinized whole bloodwere added to 5 mL of appropriate antibodies in each test-tube.The mixture was incubated in the dark at 4�C for 20 minutes.Phosphate-buffered saline (PBS 1�) (2 mL) was added to themixture, which was then centrifuged for 5 minutes 167.7 g andthe supernatant discarded. Red blood cells were lysed by adding1 mL of lysing buffer (Cell-Lyse, Becton Dickinson, San Jose,CA, USA) for 15 minutes in the dark at room temperature. PBS(2 mL) was added to the samples, centrifuged for 5 minutes167.7 g and the supernatant discarded. The pellets were resus-pended in 1 mL of PBS/formaldehyde 1% and stored at 4�C inthe dark until analysis. Flow cytometry was performed using aFACSCalibur instrument and CellQuest software (BectonDickinson, San Jose, CA, USA). Compensation parameters formultiparametric studies were adjusted at the start of analysis and10,000 events per sample were acquired with a live gate appliedto the lymphocytes using CD45. For the analysis of monocytes,we used CD14 antibodies to create the monocyte gate (BectonDickinson).

Statistical analysis

All variables were tested for normality of distribution by theShapiroeWilk test. For those that showed normality, we usedthe paired t test for comparison of baseline and post-training(mean ± standard error). For the variables that deviated fromnormality, we used the nonparametric Wilcoxon test (medianand interquartile range). The correlations between normal var-iables were performed by the Pearson correlation test and thenonparametric variables by the Spearman correlation test. Allanalyses were performed by SPSS version 19.0 (SPSS Inc.,Chicago, IL, USA). All statistical tests were two-tailed andwereperformed using a significance level of a ¼ 0.05.


//dx.doi.org/10.1016/j.jesf.2014.06.002

Table 3

Serum cytokines of participants at baseline and after 12 weeks of concurrent

training periods.

Period Baseline Post-training

hs-CRP (mg/dl) 0.14 (0.08, 0,26) 0.28 (0.23, 0.37)*

IL-6 (ng/mL) 0.36 ± 0.11 0.43 ± 0.10

TNF-a (ng/mL) 0.47 ± 0.08 1.09 ± 0.18*

INF-g (ng/mL) 0.42 ± 0.16 0.43 ± 0.17

IL-17A (ng/mL) 0.04 (0.03, 0.05) 0.05 (0.03, 0.06)

IL-10 (ng/mL) 0.52 ± 0.13 0.51 ± 0.09

Nonparametric variables are presented as median (25th centile, 75th centile),

Wilcoxon test; other variables are presented as mean ± standard error, paired t

test.

*p < 0.05 compared with baseline.

hs-CRP ¼ high-sensitive C-reactive protein; IL ¼ interleukin; INF-

g ¼ interferon-g; TNF-a ¼ tumour necrosis factor-a.


+ MODEL

Results

The 14 patients were characterized as overweight or obese(25 kg/m2 < BMI < 39.9 kg/m2), aged 35e66 years, non-smokers, and normotensive. The anthropometric charac-teristics, VO2PEAK and time to exhaustion at baseline and post-training of all participants are shown in Table 2. A signifi-cant reduction in weight (p ¼ 0.015), BMI (p ¼ 0.011), and fatmass percentage (p ¼ 0.013) was seen after the concurrenttraining. In addition, we observed a significant increase in thetime to exhaustion (p ¼ 0.017) in the cardiorespiratory exer-cise test. Changes in BMI, VO2PEAK, and time to exhaustionwere not significantly correlated (data not shown).

The pro-inflammatory cytokines (IL-6, TNF-a, IL-17A,and INF-g), anti-inflammatory IL-10, as well as the hs-CRPlevels, were analyzed. We observed increased levels of TNF-a (p ¼ 0.023) and hs-CRP (p ¼ 0.001) after 12 weeks ofconcurrent training (Table 3). The baseline values of somecytokines were positively correlated, such as IL-6 and IL-10(r ¼ 0.648, p ¼ 0.043), IL-10 and IL-17A (r ¼ 0.658,p ¼ 0.020), IL-10 and INF-g (r ¼ 0.758, p ¼ 0.011), IL-17Aand IFN-g (r ¼ 0.699, p ¼ 0.024), and TNF-a and IFN-g(r ¼ 0.661, p ¼ 0.038).

Regarding the frequency of T lymphocytes and monocytesafter the concurrent training period, we observed higher fre-quencies of CD3þCD4þ and CD3þCD8þ T cells (p ¼ 0.001and p ¼ 0.002, respectively; Fig. 1) and lower frequencies ofHLA-DRþ monocytes (p ¼ 0.021; Fig. 2).

Discussion

To our knowledge, this is the first study addressinganthropometric, inflammatory and immunological parametersof overweight and obese patients after 12 weeks of concurrenttraining. We observed a reduction in weight, BMI, waistcircumference, abdomen circumference, hip circumference,and mass percentage of fat in parallel with an increase in thetime to exhaustion after concurrent training. We also observed

Table 2

Anthropometric characteristics, VO2PEAK and time to exhaustion of participants

at baseline and after 12 weeks of concurrent training.

Period Baseline Post-training

Height (m) 1.64 ± 0.03

Weight (kg) 88.07 ± 3.16 85.11 ± 2.74*

BMI (kg/m2) 32.87 ± 0.87 31.82 ± 0.82*

WC (cm) 97.75 ± 1.78 93.81 ± 2.00*

AC (cm) 105.92 ± 1.73 102.11 ± 1.58*

HC (cm) 113.79 ± 2.01 111.03 ± 1.83

Fat mass (%) 39.0 (33.5, 40.0) 37.3 (31.5, 39.4)*

Free fat mass (%) 51.7 (48.0, 71.0) 51.6 (47.6, 70)

VO2PEAK (mL kg min�1) 27.86 ± 1.40 30.94 ± 2.52

Time to exhaustion (min) 13.75 ± 0.59 15.11 ± 0.32*

Nonparametric variables are presented as median (25th centile, 75th centile),

Wilcoxon test; other variables are presented as mean ± standard error, paired t

test.

*p < 0.05 compared with baseline.

AC ¼ abdominal circumference; BMI ¼ body mass index; HC ¼ hip

circumference; VO2PEAK ¼ peak oxygen uptake; WC ¼ waist circumference.



higher levels of TNF-a and hs-CRP after the training period.Moreover, overweight and obese patients showed increasedfrequencies of T lymphocytes (CD3þCD4þ andCD3þCD8þ) and lower proportions of monocytes (HLA-DRþ) after 12 weeks of concurrent training.

Several studies have shown that moderate physical exercisepromotes the modulation of inflammation in skeletal muscleand adipose tissue, in addition to improving physical fitnessand body measurements. The data presented here agree withprevious studies suggesting improvements in body composi-tion and aerobic capacity after aerobic or concurrent train-ing.20,25e30 In a study evaluating overweight men, Libardiet al27 observed improvements in muscle strength and physicalperformance after 16 weeks of aerobic, resistant, or concurrentexercises. The increase in the time to exhaustion observed in

Fig. 1. Basal and post-training frequencies of T circulating lymphocytes (T

helper and cytotoxic cells) of overweight and obese patients. (A) CD3þCD4þT cells; and (B) CD3þCD8þ T cells. *p < 0.05 compared with baseline

mean ± standard error, paired t test. CD ¼ cluster of differentiation.


//dx.doi.org/10.1016/j.jesf.2014.06.002

Fig. 2. Basal and post-training frequencies of HLA-DRþ (activation marker of

MHC class II) of circulating lymphocytes and monocytes of overweight and

obese patients. *p < 0.05 compared with baseline mean ± standard error,

paired t test. HLA-DR ¼ human leukocyte antigen-DR; MHC ¼ major his-

tocompatibility complex.


+ MODEL

our patients may suggest a physiological adaptation of thebody resulting from aerobic exercise, which helps to decreasemortality as well as the chance of developing cardiovasculardisease.31 Moreover, the strength training is able to increasemuscle mass as well as nutritional protein status.32,33 Aerobictraining also brings benefits such as a reduction in body fat,which can attenuate inflammation and ameliorate nutritionalstatus, contributing to reducing the waist and abdominal cir-cumferences, which are indirect measures of visceral fat.25,34

In this way, the exercise may be used with preventive andtherapeutic purposes, attenuating degenerative processes andsystemic inflammation.35

Some cytokines are produced by monocytes and/or mac-rophages in the subendothelial adipose tissue and about 30%of IL-6 is released by visceral adipose tissue.36,37 In additionto adipose tissue, skeletal muscle is also an important sourceof cytokines.38 TNF-a is involved in insulin resistance as it isoverproduced by adipose tissue in obese patients.39 CRP, anacute phase protein, is produced by the liver in response toinflammatory cytokines such as IL-6 and TNF-a and can bemodulated by exercise, depending on the intensity and dura-tion of the sessions.36,40 According to our results, higher levelsof inflammatory markers such as TNF-a and hs-CRP(p < 0.05) were observed in overweight and obese patientsafter training. Martín-Cordero et al,41 in a study evaluatingobese rats, showed that regular exercise was able to improvethe release of TNF-a by peritoneal macrophages, improvingthe balance between inflammatory markers, reaching thebehavior of lean healthy rats.

Another interesting study that evaluated overweight menafter 16 weeks of training did not observe differences incytokine levels. However, the authors indicated a change inhs-CRP levels, suggesting an increase in this variable duringthe post-training period. It is well known that CRP has anacute response to exercise and it can remain increased forseveral days following the last session of exercise.12 In ourstudy, blood was collected 48 hours after the last exercisesession, which may reflect the increased level of hs-CRP aftertraining. Furthermore, there is evidence that training pro-grams performed weekly at high frequencies can be verystressful, increasing some inflammatory markers.42 Thereforewe suggest that the short duration (3 months) of the



intervention used in our study may have induced an acutelow-grade inflammation in patients, as the progression ofexercises and the number of series/repetitions increasedrapidly, increasing the levels of inflammatory markers such asCRP and TNF-a.

Thus a long period (more than 3 months) of moderate ex-ercise could regulate inflammation. However, our datacontradict previously published studies which showed areduction in pro-inflammatory biomarkers after the perfor-mance of different types of exercise in obese and sedentarypatients, highlighting the anti-inflammatory role exerted byexercise.26,43e51 Balducci et al,26 in a study of 12 months ofexercise training in patients with type 2 diabetes mellitus andmetabolic syndrome, found decreased levels of hs-CRP, TNF-a, IL-6, and IFN-g. Jorge et al50 submitted obese type 2 dia-betic patients to aerobic, resistance, and concurrent trainingfor 12 weeks and observed reductions in the hs-CRP levels inall groups. Moreover, studies involving the effect of differentmodalities of exercise on inflammation are still controversial,requiring further research in tissue and molecular levels toclarify the correct effects of physical activities on immunityand the precise cell source of cytokines.

This study investigated the frequencies of CD3þCD4þ andCD3þCD8þ T lymphocytes as well as the percentage ofHLA-DRþ lymphocytes and monocytes in obese and over-weight patients after 12 weeks of concurrent training. There isno published data correlating the expression of CD4, CD8, andHLA-DR in peripheral blood cells with exercise in obese/sedentary patients. Fu et al52, in an animal model, have shownthat moderate exercise training prevents a decrease in CD4þ Tcells. Our study was characterized by exercise of moderateintensity (50e75% VO2PEAK) with linear progression, whichcould help to prevent the decrease in CD4þ T lymphocytes.Therefore moderate exercise appears to improve the defensemechanisms of the body by increasing the function of leuko-cytes and decreasing susceptibility to disease, whereas intenseexercise seems to weaken them.53 Shimizu et al,54 evaluating12 weeks of concurrent training in elderly patients, observedincreased frequencies of CD3þCD28þ T lymphocytes afterthe training period, indicating a high proliferation of cytotoxicand senescent T cells.

In conclusion, our data suggest that 12 weeks of concurrenttraining were able to reduce anthropometric parameters inparallel with an increase in the physical fitness of theseoverweight and obese patients. However, the progression ofexercises may have been physiologically stressful, increasingthe levels of TNF-a and hs-CRP proinflammatory biomarkers.Moreover, the concurrent training may have modulated cellimmunity by increasing CD3þCD4þ and CD3þCD8þ Tlymphocytes and decreasing HLA-DRþ monocytes. We sug-gest that the practice of regular, supervised, and moderatephysical exercise may act as a protective factor against thecomorbidities related to obesity. Further studies are needed tostandardize the appropriate type, frequency, and intensity ofexercise applied to different populations, such as overweightand obese individuals, with regard to inflammatory andimmunological biomarkers.


//dx.doi.org/10.1016/j.jesf.2014.06.002


+ MODEL

Conflicts of interest

The authors declare no potential conflicts of interest.

Acknowledgments

The authors acknowledge the financial support of CentroUniversit�ario Metodista do IPA and CAPES. They also thankBalagu�e Center Laboratory for carrying out some analysis.

References

1. Murthy N, Mukherjee S, Ray G, et al. Dietary factors and cancer che-

moprevention: an overview of obesity-related malignancies. J Postgrad

Med. 2009;55:45e54.

2. Grundy SM, Brewer Jr HB, Cleeman JI, et al, American Heart Associa-

tion, National Heart, Lung, and Blood Institute. Definition of metabolic

syndrome: report of the National Heart, Lung, and Blood Institute/

American Heart Association conference on scientific issues related to

definition. Circulation. 2004;109:433e438.

3. World Health Organization (WHO). What Is the Scale of the Obesity

Problem in Your Country? WHO Global Infobase e Data for Saving

Lives; 2010. Available from: http://www.who.int/topics/obesity/en/

[Accessed May 2011].

4. Festa A, D'Agostino Jr R, Williams K, et al. The relation of body fat mass

and distribution to markers of chronic inflammation. Int J Obes Relat

Metab Disord. 2001;25:1407e1415.

5. Bullo M, Garcia-Lorda P, Megias I, et al. Systemic inflammation, adipose

tissue tumor necrosis factor, and leptin expression. Obes Res.

2003;1:525e531.

6. Bastard JP, Jardel C, Bruckert E, et al. Elevated levels of interleukin 6

are reduced in serum and subcutaneous adipose tissue of obese

women after weight loss. J Clin Endocrinol Metab. 2000;85:

3338e3342.

7. Yudkin JS, Stehouwer CD, Emeis JJ, et al. C reactive protein in healthy

subjects: associations with obesity, insulin resistance, and endothelial

dysfunction: a potential role for cytokines originating from adipose tissue?

Arterioscler Thromb Vasc Biol. 1999;19:972e978.

8. Dusserre E, Moulin P, Vidal H. Differences in mRNA expression of the

proteins secreted by the adipocytes in human subcutaneous and visceral

adipose tissues. Biochim Biophys Acta. 2000;1500:88e96.

9. Weiseberg SP, McCann D, Desai M, et al. Obesity is associated with

macrophage accumulation in adipose tissue. J Clin Invest.

2003;112:1796e1808.

10. Esposito K, Pontillo A, Di Palo C, et al. Effect of weight loss and lifestyle

changes on vascular inflammatory markers in obese women: a randomized

trial. JAMA. 2003;289:1799e1804.

11. Nicklas BJ, You T, Pahor M. Behavioral treatments for chronic systemic

inflammation: effects of dietary weight loss and exercise training. Can J

Appl Physiol. 2005;172:1199e1209.

12. Kasapis C, Thompson PD. The effects of physical activity on serum C-

reactive protein and inflammatory markers: a systematic review. J Am Coll

Cardiol. 2005;45:1563e1569.

13. Simonson SR. The immune response to resistance exercise. J Strength

Cond Res. 2001;15:378e384.

14. Ischander M, Zaldivar F, Eliakim A, et al. Physical activity, growth, and

inflammatory mediators in BMI-matched female adolescents. Med Sci

Sports Exerc. 2007;39:1131e1138.15. Pischon T, Hankinson S, Hotamisligil S, et al. Leisuretime physical ac-

tivity and reduced plasma levels of obesity-related inflammatory markers.

Obes Res. 2003;11:1055e1064.

16. Straczkowski M, Kowalska I, Dzienis-Straczkowska S, et al. Changes in

tumor necrosis factor a system and insulin sensitivity during an exercise

training program in obese women with normal and impaired glucose

tolerance. Eur J Endocrinol. 2001;145:273e280.



17. Ryan AS, Pratley RE, Elahi D, et al. Changes in plasma leptin and insulin

action with resistive training in postmenopausal women. Int J Obes Relat

Metab Disord. 2000;24:27e32.

18. Miller JP, Pratley RE, Goldberg AP, et al. Strength training increases

insulin action in healthy 50- to 65-year-old men. J Appl Physiol.

1994;77:1122e1127.

19. Zachwieja JJ, Toffolo G, Cobelli C, et al. Resistance exercise and growth

hormone administration in older men: effects on insulin sensitivity and

secretion during a stable-label intravenous glucose tolerance test. Meta-

bolism. 1996;45:254e260.

20. Kelley GA, Kelley KS. Effects of aerobic exercise on c reactive protein,

body composition, and maximum oxygen consumption in adults: a meta-

analysis of randomized controlled trials.Metabolism. 2006;55:1500e1507.

21. Bruce RA. Exercise testing of patients with coronary heart disease.

Principles and normal standards for evaluation. Ann Clin Res.

1971;3:323e332.22. Borg GAV. Psychophysical bases of perceived exertion. Med Sci Sports

Exerc. 1982;14:377e381.

23. Brown LE, Weir JP. ASEP procedures recommendations I: accurate

assessment of muscle strength and power. J Exerc Physiol Online.

2001;4:1e21.

24. American College of Sports Medicine (ACSM). ACSM's Guidelines for

Exercise Testing and Prescription. 8th ed. Baltimore, MD: The Point;

2009.

25. Donges CE, Duffield R, Drinkwater EJ. Effects of resistance or aerobic

exercise training on interleukin-6, c-reactive protein, and body composi-

tion. Med Sci Sports Exerc. 2010;42:304e313.26. Balducci S, Zanuso S, Nicolucci A, et al. Anti-inflammatory effect of

exercise training in subjects with type 2 diabetes and the metabolic syn-

drome is dependent on exercise modalities and independent of weight

loss. Nutr Metab Cardiovasc Dis. 2010;20:608e617.

27. Libardi CA, Souza GV, Cavaglieri CR, et al. Effect of resistance, endur-

ance, and concurrent training on TNF-a, IL-6, and CRP. Med Sci Sports

Exerc. 2012;44:50e56.28. Aicher BO, Haser EK, Freeman LA, et al. Diet-induced weight loss in

overweight or obese women and changes in high-density lipoprotein levels

and function. Obesity. 2012;20:2057e2062.

29. Fisher G, Hyatt TC, Hunter GR, et al. Effect of diet with and without

exercise training on markers of inflammation and fat distribution in

overweight women. Obesity (Silver Spring). 2011;19:1131e1136.

30. Melo CM, Tirapegui J, Cohen D, et al. Nutritional status and energy

expenditure after a programme of nutrition education and combined aer-

obic/resistance training in obese women. E Spen Eur E J Clin Nutr Metab.

2010;5:e180ee186.

31. Blair SN, Kampert JB, Kohl HW, et al. Influences of cardiorespiratory

fitness and other precursors on cardiovascular disease and all-cause

mortality in men and women. JAMA. 1996;276:205e210.

32. Donnelly JE, Blair SN, Jakicic JM, et al. Appropriate physical activity

intervention strategies for weight loss and prevention of weight regain for

adults. Med Sci Sports Exerc. 2009;41:459e471.

33. Gibson R. Principles of Nutritional Assessment. 1st ed. New York, NY:

Oxford University Press; 1990, 690 pp.

34. Wajchenberg BL. Subcutaneous and visceral adipose tissue: their relation

to the metabolic syndrome. Endocr Rev. 2000;21:697e738.

35. Prestes J, Shiguemoto G, Botero JP, et al. Effects of resistance training on

resistin, leptin, cytokines, and muscle force in elderly post-menopausal

women. J Sports Sci. 2009;27:1607e1615.

36. Petersen AM, Pedersen BK. The anti-inflammatory effect of exercise. J

Appl Physiol. 2005;98:1154e1162.

37. Mohamed-Ali V, Pinkney JH, Coppack SW. Adipose tissue as an endo-

crine and paracrine organ. Int J Obes Relat Metab Disord.

1998;22:1145e1158.

38. Steensberg A, Keller C, Starkie RL, et al. IL-6 and TNF-a expression in,

and release from, contracting human skeletal muscle. Am J Physiol

Endocrinol Metab. 2002;283:E1272eE1278.

39. Hotamisligil GS, Arner P, Caro JF, et al. Increased adipose tissue

expression of tumor necrosis factor-alpha in human obesity and insulin

resistance. J Clin Invest. 1995;95:2409e2415.


//dx.doi.org/10.1016/j.jesf.2014.06.002

http://www.who.int/topics/obesity/en/


+ MODEL

40. Baumann H, Gauldie J. The acute phase response. Immunol Today.

1994;15:74e80.

41. Martín-Cordero L, García JJ, Hinchado MD, et al. Habitual physical ex-

ercise improves macrophage IL-6 and TNF-a deregulated release in the

obese zucker rat model of the metabolic syndrome. Neuro-

immunomodulation. 2011;18:123e130.

42. de Salles BF, Sim~ao R, Fleck SJ, et al. Effects of resistance training on

cytokines. Int J Sports Med. 2010;31:441e450.

43. Andersson J, Janssona J, Hellsten G, et al. Effects of heavy endurance

physical exercise on inflammatory markers in nonathletes. Atheroscle-

rosis. 2010;209:601e605.

44. Ahmadi N, Eshaghian S, Huizenga R, et al. Effects of intense exercise and

moderate caloric restriction on cardiovascular risk factors and inflam-

mation. Am J Med. 2011;124:978e982.

45. Snel M, van Diepen JA, Stijnen T, et al. Immediate and long-term effects

of addition of exercise to a 16-week very low calorie diet on low-grade

inflammation in obese, insulin-dependent type 2 diabetic patients. Food

Chem Toxicol. 2011;49:3104e3111.

46. El-Kader SMA. Aerobic versus resistance exercise training in modulation

of insulin resistance, adipocytokines and inflammatory cytokine levels in

obese type 2 diabetic patients. J Adv Res. 2011;2:179e183.

47. Smart NA, Larsen AI, LeMaitre JP, et al. Effect of exercise training on

interleukin-6, tumour necrosis factor alpha and functional capacity in

heart failure. Cardiol Res Pract. 2011;27:532e620.



48. Buyukyazi G, Ulmanb C, Taneli F, et al. The effects of different intensity

walking programs on serum blood lipids, high sensitive C reactive protein,

and lipoprotein associated phospholipase A2 in premenopausal women.

Science Sports. 2010;25:245e252.

49. Jae SY, Fernhalla B, Heffernan KS, et al. Effects of lifestyle modifications

on C-reactive protein: contribution of weight loss and improved aerobic

capacity. Metabolism. 2006;55:825e831.

50. Jorge MLMP, Oliveira VN, Resende NM, et al. The effects of aerobic,

resistance, and combined exercise on metabolic control, inflammatory

markers, adipocytokines, and muscle insulin signaling in patients with

type 2 diabetes mellitus. Metabolism. 2011;60:1244e1252.

51. Shih KC, Janckila AJ, Kwok CF, et al. Effects of exercise on insulin

sensitivity, inflammatory cytokines, and serum tartrate-resistant acid

phosphatase 5a in obese Chinese male adolescents. Metabolism.

2010;59:144e151.

52. Fu SC, Qin L, Leung CK, et al. Regular moderate exercise training pre-

vents decrease of CD4þ T-lymphocytes induced by a single bout of

strenuous exercise in mice. Can J Appl Physiol. 2003;28:370e381.

53. da Nobrega AC. The subacute effects of exercise: concept, characteristics,

and clinical implications. Exerc Sport Sci Rev. 2005;33:84e87.

54. Shimizu K, Suzuki N, Imai T, et al. Monocyte and T-cell responses to

exercise training in elderly subjects. J Strength Cond Res.

2011;25:2565e2572.


//dx.doi.org/10.1016/j.jesf.2014.06.002

Effects of concurrent training on inflammatory markers and expression of CD4, CD8, and HLA-DR in...

Documents

Transcript of Effects of concurrent training on inflammatory markers and expression of CD4, CD8, and HLA-DR in...