QOPNA
Departamento de Química
Universidade de Aveiro
BOOK OF ABSTRACTS
rd
3 WORKSHOP OF
IMMUNOLOGY
th
20 MAY 2015 – UNIVERSITY OF AVEIRO
PROGRAM
13:45
Registration
14:15
Bruno Neves, QOPNA UA
Vacinas de Células Dendríticas no combate ao cancro: estado atual e perspetivas futuras
14:45
Alexandre Salvador, ENZIfarma S.A.
Resposta imunológica monitorizada por citometria de fluxo
15:15
Virgínia Carvalhais, CBE UM
Immunoproteomics to identify Proteins involved in Host-Pathogen Interaction: Example of
Staphylococcus epidermidis Biofilms
15:30
Elisabete Maciel, QOPNA UA
Oxidized phosphatidylserine downregulate inflammatory signals in macrophages
15:45
Ana Campos, QOPNA UA and Cell 2B.
Lipidomics of Mesenchymal Stromal Cells, powerful immunosuppressive agents
16:15
Coffee break funded by Reagente 5
16:45
Andreia Monteiro, FCS UBI
O linfócito T na Esclerose Múltipla
17:15
João Martins, CNC UC
Dendritic cells surrogates for chemicals’ allergenecity screenning
17:30
Marta Duque, QOPNA UA
The Immunosuppresive Potential of Human Amniotic Membrane Extract
17:45
Luciele Minuzzi, FCDEF UC
Estudo da senescência em células T reguladoras de desportistas seniores
18:15
Closing ceremony
ORGANIZING COMMITTEE
Rosário Domingues
Pedro Domingues
Elisabete Maciel
Cláudia Simões
Ana Reis
João Martins
Rui Vitorino
Núcleo de estudantes de Química
Mass Spectrometry Center, QOPNA, Department of Chemistry, University of Aveiro
3rd Workshop of Immunology
VACINAS DE CÉLULAS DENDRITICAS NA TERAPIA ANTI TUMORAL: ESTADO
ATUAL E PERSPETIVAS FUTURAS
NEVES B.M.
1 Faculty
of Pharmacy and Centre for Neuroscience and Cell Biology, University of Coimbra 3000-548,
Coimbra – Portugal
2 Mass Spectrometry Centre, Department of Chemistry and QOPNA, University of Aveiro, Campus
Universitário de Santiago, 3810-193 Aveiro – Portugal
E-mail: [email protected]
Abstract
As células dendríticas (CDs) são células apresentadoras de antigénio com um papel central na
interligação entre imunidade inata e adaptativa. Possuem uma capacidade inigualável de modular as
respostas imunes, sendo responsáveis quer pela indução de imunidade quer pela manutenção de
tolerância a auto antigénios. Por esta razão tornaram-se nas últimas duas décadas alvos
preferenciais no desenvolvimento de imunoterapias, nomeadamente nas que se centram no combate
ao cancro. Os resultados experimentais extremamente promissores nesta área não foram muitas das
vezes acompanhados por similar sucesso na transposição para aplicação clinica. Este facto levou a
que se instalasse alguma desconfiança acerca do real valor terapêutico das vacinas anti tumorais
baseadas em CDs. No entanto, o aumento do nosso conhecimento acerca da imunobiologia destas
células e o estabelecimento das características ótimas para a imunoterapia anti tumoral, permitiram
nos últimos anos desenvolver novas abordagens de uma forma muito mais racional. Contrariamente
aos primeiros ensaios clínicos que testavam as vacinas de CDs em monoterapia, atualmente estas
são combinadas com outros tratamentos, nomeadamente quimioterapia e radioterapia, no intuito de
se obter um efeito sinergístico. Em termos de desenvolvimento muito está ainda por fazer, no entanto
novas abordagens como por exemplo o direcionamento in vivo e o uso de implantes com capacidade
para recrutar, carregar e maturar as CDs, afiguram-se como tecnologias bastante promissoras que
irão num futuro próximo afirmar-se como armas efetivas no combate ao cancro.
Aveiro | 20th May 2015
3rd Workshop of Immunology
RESPOSTA IMUNOLÓGICA MONITORIZADA POR CITOMETRIA DE FLUXO
Salvador A. 1,2
1 Enzifarma
S.A
2 ESTES-L
E-mail: [email protected]
Abstract
Sendo a citometria de fluxo uma metodologia que permite a análise individual e individualizada de
células em suspensão, rapidamente se percebeu a utilidade da mesma, na interpretação do papel de
cada célula no contexto do órgão e do individuo. Assim, cada célula tem interações com as demais
células e neste contexto, a forma como recebe informação ou estimulo, desencadeia em si uma
resposta. Esta resposta vai condicionar, moldar ou potenciar a resposta de outras células. Disto o
sistema imunológico é um bom exemplo.
Normalmente podemos estimular as células do sistema imunológico “in vitro” de forma a verificarmos
a capacidade de resposta a um estímulo. Com a simples estimulação com Ionomicina ou PMA,
podemos verificar a capacidade de resposta de um determinado individuo e comparar com a resposta
“normal”. Com a citometria de fluxo, podemos identificar o tipo de interleucina ou citoquina que cada
célula está a produzir. Também com a citometria, se pretendermos, podemos quantificar as citocinas
que são libertadas para o meio extra-celular.
Sabendo que, as citocinas ou interleucinas ao serem libertadas, vão produzir um efeito ao se ligarem
a recetores celulares específicos, podemos ainda usar a citometria para responder às perguntas: que
citocina estimula cada célula? Que quantidades de citocina são necessárias? O estímulo é específico,
ou existem mais células que respondem ao mesmo estímulo?
Até hoje, e por limitações destas metodologias, como sensibilidade e reprodutibilidade, a
quantificação de uma proteína por célula era apenas possível por outras técnicas. Hoje em dia
podemos dizer que estimulo sofreu, de que célula se trata e ainda dizer qualitativamente ou de forma
quantitativa a(s) proteína(s) que tiverem intervenção no processo de sinalização celular.
Com o objetivo de verificar a resposta imunológica, pretendemos com a citometria de fluxo, identificar
as células que participam nessa resposta, que citocinas ou interleucinas são secretadas e com que
células interagem. Será que a resposta imunológica ou a sua intensidade é dependente de todo este
processo?
A metodologia está ai! Os reagentes e protocolos estão disponíveis! Será apenas necessário obter
amostras e ter imaginação! O espirito científico, aliado à nossa curiosidade e necessidade de
compreensão dos processos biológicos leva-nos a crer que a citometria é uma das melhores formas
para compreender a resposta imunológica.
Aveiro | 20th May 2015
3rd Workshop of Immunology
IMMUNOPROTEOMICS TO IDENTIFY PROTEINS INVOLVED IN HOSTPATHOGEN INTERACTION: EXAMPLE OF Staphylococcus epidermidis
BIOFILMS
CARVALHAIS V1,2, CERVEIRA F3, VILANOVA M4, CERCA N2, VITORINO R1,5
1
QOPNA, Mass Spectrometry Center, Department of Chemistry, University of Aveiro, 3810-193,
Aveiro, Portugal
2 CEB - Centre of Biological Engineering, LIBRO - Laboratory of Research in Biofilms Rosário Oliveira,
University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal
3 Anatomia Patológica, Centro Hospitalar Baixo-Vouga, Avenida Artur Ravara, 3814-501 Aveiro,
Portugal.
4 Laboratory of Immunology, Department of Immuno-Physiology and Pharmacology, Institute of
Biomedical Sciences Abel Salazar (ICBAS), University of Porto, 4050-313 Porto, Portugal
5 iBiMED, Institute for Biomedical Research, University of Aveiro, Aveiro, Portugal
E-mail: [email protected]
Abstract
Immunoproteomics allows the identification of immunogenic and immunoreactive proteins that may
participate in host-pathogen interactions and in the host immune response [1]. Therefore,
immunoproteomics improves the understanding of infection pathogenesis and unravels novel
therapeutic targets based on the repertoire of immunogens [2,3]. Here, we intended to identify
immunogenic proteins of Staphylococcus epidermidis biofilms associated with dormancy condition. For
that, we separated whole proteins from S. epidermidis biofilms with induced and prevented dormancy
using two-dimensional electrophoresis (2-DE). Then, immunoblotting was performed with human sera
and proteins were identified by mass spectrometry (MS). A total of 19 bacterial proteins recognized by
human serum samples were identified. Differences in the reactivity pattern were found between
biofilms with prevented and induced dormancy. CodY and GpmA proteins were reactive to sera only
when biofilm dormancy was induced, while FtnA and ClpP were only reactive when dormancy was
prevented. Further evaluation of these proteins can provide insights into the mechanisms related to
dormancy and help to improve current understanding on how dormancy affects the host immune
response. These proteins may represent promising candidates as biofilm markers allowing
discrimination of physiological condition displayed by biofilm bacteria and will be worth to consider in
further studies.
References
[1] Fulton, KM. et al. Immunoproteomics: current technology and applications. Methods Mol. Biol.
2013, 1061, 21
[2] Dennehy, R. et al. Immunoproteomics: the key to discovery of new vaccine antigens against
bacterial respiratory infections. Curr Protein Pept Sci., 2012, 13, 807
[3] Fulton, KM. et al. Methods and applications of serological proteome analysis. Methods Mol. Biol.
2013, 1061, 97
Aveiro | 20th May 2015
3rd Workshop of Immunology
OXIDIZED
PHOSPHATIDYLSERINE
SIGNALS IN MACROPHAGES
DOWNREGULATE
INFLAMMATORY
MACIEL E.1, MARTINS J.D.2, NEVES B.M. 1, CRUZ M.T. 2, DOMINGUES P. 1, DOMINGUES
M.R1
1 Mass
Spectrometry Centre, Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal
Faculty of Pharmacy and Center for Neuroscience and Cell Biology (CNC), University of Coimbra,
3000-548 Coimbra, Portugal
E-mail: [email protected]
2
Abstract
In the last years phosphatidylserine (PS) has been the focus of many studies and some antiinflammatory properties were attributed to this phospholipid. The development of analytical methods
for lipid hydroperoxides has revealed the important role of oxidized PS (oxPS) during apoptosis as
well as the novel roles of oxPS under pathophysiological conditions [1].
The present work aimed to clarify whether PS and oxPS differentially modulate the functional
responses of macrophages by evaluating the influence of oxPS on the macrophages inflammatory
status. Thus, we determined the effects of oxidized 1-palmitoyl-2-linoleoyl-phosphatidylserine
(oxPLPS) and PLPS on the RAW 264.7 macrophages production of the pro-inflammatory mediator
nitric oxide (NO) and on the levels of the inducible NO synthase (Nos2) and Il1 mRNA. The ability of
PLPS and oxPLPS to modulate the lipopolysaccharide (LPS)-triggered macrophage activation was
also analyzed. Finally, the effects of PLPS species over canonical inflammation-associated signaling
pathways, such as nuclear factor (NF)-ĸB and mitogen-activated protein kinases (MAPKs) were also
disclosed.
The results obtained showed that both PLPS and oxPLPS species are deprived of intrinsic proinflammatory activity. However, only oxPLPS were found to significantly inhibit NO production and
iNos and IL1β genes transcription induced by LPS. At a molecular level, these effects were partially
due to attenuation of LPS-induced c-Jun-N-terminal kinase (JNK) phosphorylation and p65 NF-ĸB
nuclear translocation. Overall our data suggest that oxPLPS, but not native PLPS, mitigates proinflammatory signaling in macrophages, contributing to containment of inflammation during apoptotic
cell engulfment.
References
[1] Matsura T. Oxidized Phosphatidylserine: Production and Bioactivities. Yonago Acta medica
2014;57:119–127.
Aveiro | 20th May 2015
3rd Workshop of Immunology
LIPIDOMICS
OF
MESENCHYMAL
IMUNOSSUPRESSIVE AGENTS
STROMAL
CELLS,
POWERFUL
CAMPOS A.1, MACIEL E.1, MOREIRA A.1, MELO T.1, DOMINGUES P.1, DOMINGUES M.1,
SANTOS F.2, CURADO L.2, ANTUNES B.2
1
Mass Spectrometry Centre, QOPNA, Department of Chemistry, University of Aveiro, 3810-193
Aveiro, Portugal
2 Cell2B Advanced Therapeutics SA, Biocant Park Núcleo 04 Lote 4 A, 3060-197 Cantanhede,
Portugal
E-mail: [email protected]
Abstract
In the last years, mesenchymal stromal cells (MSCs) have been successfully applied in many clinical
trials (1). MSCs are primitive cells with origin in the mesodermal germ layer. They are considered adult
stem cells, once these cells are characterized as capable of self-renewal and directed differentiation
(1).
MSCs have drawn the interest of researchers and they have been used as a biologic therapeutic for
several clinical applications (1). In fact, MSCs interfere with immune cells and inhibit the proliferation of
activated allogeneic lymphocytes either in vitro as in vivo, crossing histocompability barriers. MSCs
also act upon most major components of the innate and adaptive immune system and they show
proangiogenic, cytoprotective and antifibrotic effects. In addition, MSCs have proven capable of
migrating towards tissues experiencing inflammatory events (2). For these reasons, most recently
there has been a rising interest in applying these cells as a treatment for inflammatory and
autoimmune diseases (2), such as Graft versus Host Disease (GvHD), multiple sclerosis and Crohn’s
disease (CD).
However, despite the wide use of these cells in clinical trials the mechanisms employed by these cells
are not totally understood. Further and more extensive applications of MSCs in other inflammatory and
autoimmune diseases require a deeper knowledge about their behaviour, phenotypic characteristics
and mechanisms. Lipidomics approach can provide some of the needed information. Lipidomics refers
to ‘the systems-level analysis of lipids and their interacting moieties’. There has been an increasing
interest in Lipidomics once each lipid class has specific functions and take part in different metabolic
and signalling processes. Moreover, it is know accepted lipids trigger immune responses. So it
becomes important to analyse differences of specific lipid species in different pathways.
Only a few and very recent studies around MSCs’ lipidomics have been conducted. However, none of
them analysed differences in MSCs’ phospholipid profile between MSCs subjected to proinflammatory stimulus and non-stimulated MSCs. In this study, we cultured MSCs both in a standard
culture medium and in a culture medium supplemented with pro-inflammatory stimulus, namely TNF-
and IFN-. Then we analysed differences in the phospholipid profile of MSCs from both conditions, in
order to understand which phospholipid molecular species take part in MSCs’ immunosuppressive
mechanism.
References
[1] Salem HK, Thiemermann C. Mesenchymal Stromal Cells : Current Understanding and Clinical
Status. Stem Cells. 2010;28(3):585–96.
[2] Griffin MD, Elliman SJ, Cahill E, English K, Ceredig R, Ritter T. Concise Review : Adult
Mesenchymal Stromal Cell Therapy for Inflammatory Diseases : How Well Are We Joining the
Dots ? Stem Cells. 2013;31(10):2033–41.
Aveiro | 20th May 2015
3rd Workshop of Immunology
THE T CELL IN MULTIPLE SCLEROSIS
MONTEIRO A. 1, CRUTO C.2, ROSADO P.2, FONSECA M.1, ROSADO L.2, CARVALHEIRO
T. 3, PAIVA A. 3
1 Faculdade
Ciências da Saúde-Centro de Investigação em Ciências da Saúde, Universidade da Beira
Interior, Covilhã
2 Serviço Neurologia, Centro Hospitalar Cova da Beira, Covilhã 3 Laboratório citometria de fluxo,
Instituto Português do Sangue e da Transplantação, Coimbra, Portugal
E-mail: [email protected]
Abstract
Multiple sclerosis (MS) is a complex autoimmune demyelinating disease of the central nervous system
(CNS) of unknown etiology. MS is characterized by CNS inflammation, demyelination, axonal injury
and axonal loss. Autoreactive T cells activated outside the CNS cross the blood-brain barrier and are
reactivated by local antigen-presenting cells. Secretion of proinflammatory cytokines stimulates
microglial cells and astrocytes, recruits additional inflammatory cells, and induces antibody production
by plasma cells. This inflammatory process finally leads to tissue damage within the plaque. It is
generally accepted that myelin-specific Th cells play a central role in initiating and orchestrating CNS
inflammation in MS. Initially, Th1 cells where considered the subset responsible for the MS
pathogenesis, although others subsets has been shown to be also involved in this process, like Th17,
regulatory Th (Treg) and γσT cells. Considerable evidence exist that points toward an important
pathogenic and/or regulatory role for cytotoxic CD8 T cells(Tc) in MS. In this context, the aim of the
present work was to quantify and functional characterize Th(c)1, Th(c)17, Treg and γσT cells
populations in the peripheral blood from patients diagnosed with MS, according to the MacDonald
criteria 2010. 38 patients, subdivided in 2 subgroups, according to the phase of the disease, stable
(n=30, 26 female/ 4 male, mean age 41±15) or relapse (n=8, 5 female/ 3male, mean age 44±11) and a
control group (n=20, 16female/4male, mean age 50±9) were evaluated in this study. Th(c)1, Th(c)17
and Tγσ cells characterization was done by intracellular cytokine staining after in vitro stimulation with
PMA/ionomycin, in the presence of Brefeldin A during 4 hours. The expression of cell surface markers
and intracellular production of IL-17, TNFα, IFNγ and IL-2 were assessed by flow cytometry
(FACSCanto; BD), and the obtained results were analyzed using Infinicyt software (Cytognos).
Statistical evaluation of data were analysed using the non-parametric Mann-Whitney U test. We
observed a significant increase in the frequency of Th17 cells in stable MS group when compared with
relapse or control groups. Moreover, an increased expression of TNFα was observed in those cells.
The frequency of Tc17cells, and among them, those that concomitant produced IL-17 and IFN-γ were
also increased in stable MS, as well as TNFα expression. A same pattern was observed in relapse MS
when compared with control group. Concerning the frequency of Th1, Tc1 and Treg cells no
statistically significant differences were observed between the studied groups. In the stable group of
patients with RRMS naive γσT were increased when compared with control group. The patients in
relapse phase only exhibit statistically significant differences when compared with stable group,
namely a decreased in effector γσT subset and an increase in the frequency of γσT producing IFNγ.
In summary, it seems that Th17, Tc17 and γσT cells could play an important role in the
pathophysiology of MS, since increased frequencies of those cells were observed in MS patients. We
also may speculate that the decreased frequencies of those cells observed in relapse patients could
reflect a specific migration from blood to CNS. We believe that a more comprehensive knowledge of
the plasticity of these cells in MS is highly pertinent for future therapeutic interventions.
References
[1] Kamm CP, Uitdehaag BM, Polman CH. Multiple Sclerosis: Current Knowledge and Future
Outlook. Eur Neurol. 2014;72:132–41.
[2] Raphael I, Nalawade S, Eagar TN, Forsthuber TG. Cytokine T cell subsets and their signature
cytokines in autoimmune and inflammatory diseases. Cytokine. Elsevier Ltd; 2014;
[3] Maddur MS, Miossec P, Kaveri S V., Bayry J. Th17 cells: Biology, pathogenesis of autoimmune
and inflammatory diseases, and therapeutic strategies. Am J Pathol. 2012;181(1):8–18.
Aveiro | 20th May 2015
3rd Workshop of Immunology
DENDRITIC CELLS
SCREENING
SURROGATES
FOR
CHEMICALS’
ALLERGENICITY
MARTINS J. D. 1,2, SILVA A. 2, MACIEL E. A. 3, FERREIRA I. 2, DOMINGUES P. 3, NEVES
B. M. 1,3, DOMINGUES M. R. M. 3, CRUZ M. T. 1,2
1 Faculty
of Pharmacy University of Coimbra, 3000-548 Coimbra, Portugal
for Neuroscience and Cell Biology (CNC), University of Coimbra, 3004-517 Coimbra, Portugal
3 Mass Spectrometry Centre, Chemistry Department, University of Aveiro, 3810-193 Aveiro, Portugal
E-mail: [email protected]
2 Center
Abstract
Dermal and inhalation toxicity are important toxicological endpoints that should be determined to all
chemicals for which human exposure is foreseeable. European Union legislations such as REACH
(Registration, Evaluation, Authorisation and Restriction of Chemicals; EC nº 1907/2006) and the
cosmetics regulation (EC nº 1223/2009) have reinforced that obligation and strongly encouraged the
reduction/avoidance of animals’ use in toxicity tests.
Allergic contact dermatitis and respiratory allergies caused by low molecular weight electrophilic
reactive chemicals are particular cases of dermal and inhalation toxic reactions, respectively. The
complexity of these adaptive immune reactions hinders the in vitro determination of chemicals’
propensity to trigger such reactions. One promising strategy towards the development non-animal
alternative approaches relies on the utilization of in vitro models based on dendritic cells (DC)
surrogates. In such models, the crucial role played by DC in the orchestration of T-cell mediated
immune responses is used as a premise to predict the allergenic potential of chemicals.
In our presentation we will briefly highlight the common and unique immune features involved in
contact and respiratory allergies triggered by chemicals and shortly review the utilization of ‘DC like’
cells in toxicological predictive tests for skin and airway sensitization. We will also present some of the
results obtained in our lab with the DC-like cell line THP-1, a dendritic cell surrogate thoroughly used
in the above mentioned toxicity tests.
Funding
This work is funded by The Johns Hopkins Center for Alternatives to Animal Testing [2014-07], by
FEDER funds through the Operational Programme Competitiveness Factors - COMPETE and Quadro
de Referência Estratégico Nacional, and by national funds by FCT -Foundation for Science and
Technology under the projects QREN – Empresas em co-promoção project ref: 38977, PEstC/SAU/LA0001/2013 and strategic projects UID / NEU / 04539 / 2013, REDE/1504/REM/2005 (that
concerns the Portuguese Mass Spectrometry Network), and the projects PEst-C/QUI/UI0062/2013
and FCOMP-01-0124-FEDER-037296 (that concern QOPNA research unit), and the fellowships
SFRH/BD/73065/2010 and SFRH/BPD/104165/2014.
Aveiro | 20th May 2015
3rd Workshop of Immunology
THE IMMUNOSUPPRESIVE POTENTIAL OF HUMAN AMNIOTIC MEMBRANE
EXTRACT
DUQUE M.1, JESUS M.2, SILVA I2, MAMEDE AC3,4,5,6, LARANJO M3,5,6, CARVALHO MJ3,5,6,7,
MOURA P7, ABRANTES AM 3,5,6, MAIA CJ4, ROSÁRIO D.1, BOTELHO MF3,5,6. TRINDADE H2,
PAIVA A.2
1Department
of Chemistry, University of Aveiro, Aveiro, Portugal
and Transplantation Center of Coimbra, Portuguese Institute of the Blood and Transplantation,
Coimbra, Portugal
3Biophysics Unit, Faculty of Medicine, University of Coimbra, Coimbra, Portugal
4CICS-UBI, Health Sciences Research Center, University of Beira Interior, Covilhã, Portugal
5CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal
6CNC-IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal
7Obstetrics Service, Coimbra Hospital and University Center, Coimbra, Portugal
E-mail: [email protected]
2Blood
Abstract
Introduction: The human amniotic membrane (hAM) is thought to possess an active
immunosuppressive mechanism responsible for its ability to inhibit innate and adaptive immune
responses, including T cell proliferation and cytokine production. Several hAM-derived factors,
potentially responsible, have already been identified but the exact underlying mechanism of action is
yet unknown. Furthermore, there is little information regarding the exact influence over individual T cell
subpopulations. Thus the proposed objective was to evaluate the influence of hAM extract over the
production of different inflammatory cytokines by different T helper (Th or CD4) and T cytotoxic (Tc or
CD8) cell subsets.
Methods: Peripheral blood mononuclear cells (MNC) were cultured during 24 hours, with or without
(negative control) the addition of hAM extract to the culture medium, and then stimulated with phorbol
12-myristate 13-acetate (PMA) for the analysis of intracellular cytokine production by flow cytometry.
The frequency of IL-2, IL-9 IL-17, TNFα and IFNγ-producing Th (CD3+CD4+CD8-) and Tc (CD3+CD4CD8+) cells were determined among naïve (CD45RA+CD27+), CM (CD45RA-CD27+), EM (CD45RACD27-) and effector (CD45RA+CD27-) T cell compartments. Evaluation of cell proliferation was
performed after mitogen stimulation, through thymidine incorporation assay, either in presence or
absence of extract.
Results: Overall, T cells showed lower frequencies of cytokine producing cells in the presence of hAM
extract. The percentage of inhibition varied according to the cytokine understudy and T cell
compartment, still, significant inhibition was observed for all cytokine-producing cells, with the possible
exception of IFNγ-producing CD8 T effector cells. For the most part, the percent inhibition was very
similar between CD4 and CD8 cells, however, CD4 TNFα- and IFNγ-producing cells were clearly more
susceptible to inhibition than the CD8 cells. On the other hand, a strong inhibitory effect over cell
proliferation was observed for PHA-stimulated cells, being slightly lower for PWM and absent for SpAstimulated cells. Additionally, the same pattern of experiments were performed with amniotic
membrane derived cells, in a ratio MNC-to-hAM cells of 10:1 and 2:1, although a significant reduction
in the frequency of cytokine producing cells was only observed for IL-2 producing CD4 and CD8 T
cells.
Conclusions: hAM extract exhibits a strong potential to suppress T cell activation and proliferation. On
the other hand, hAM cells do not exert the same effect, which could be due, at least in part, to the
lower frequency of mesenchymal stromal cells detected in cells suspensions from amniotic
membrane.
Aveiro | 20th May 2015
3rd Workshop of Immunology
ACUTE EXERCISE EFFECTS ON TREG CELLS IN MASTER ATHLETES
MINUZZI L.1, RAMA L.1, ROSADO F.1, PEREIRA S.2, SILVA I.2, INACIO M.2, PAIVA A.2,
TEIXEIRA A.1
1 Faculty
of Sport Sciences and Physical Education, University of Coimbra, Coimbra, PORTUGAL
Portuguese Institute for Blood and Transplantation, São Martinho do Bispo, Coimbra, PORTUGAL
[email protected]
2
Abstract
Human aging is associated with a progressive decline in immune function. Among these changes are
alterations in T-cell subpopulations size, cytokine secretion pattern, cell replicative capacity and
antibody production, all of which culminate in a pro-inflammatory state and diminished capacity to
respond to new antigens, alterations are closely related to the increased mortality and morbidity rates
observed in this population (1). A feature of immunosenescence is the change in the cellular
composition of the T-cell compartment, including a decrease in the number of naïve T cells and an
increase in the number of memory T cells. T-cells that have reached terminal differentiation are known
to have overly eroded and critically short telomeres (2). These erode progressively with each round of
cell division as in T-cell clonal expansion as in response to persistent viral infections. Excessive
telomere erosion triggers mechanistic pathways for senescence causing the cell to undergo
proliferative arrest. Elderly individuals are known to have shorter leukocyte telomeres than their
younger counterparts (3). However, analysis of human peripheral blood after exercise has revealed an
increase in regulatory T cells (Tregs) (4). These are CD4+CD25+ cells that express the transcription
factor FOXP3, the main function appearing to be the downregulation of adaptative immune responses
(5) and the minimization of inflammation-induced tissue damage. Studies have shown that antigen
stimulated IL-10 production highest in the athletes with the higher training loads (6). In trained rats,
high intensity exercise resulted in increased numbers of regulatory T cells, and was also associated
an increased expression of anti-inflammatory cytokines (7). These results imply that high intensity
exercise training may be more beneficial than moderate training in reducing risk of cardiovascular and
metabolic chronic diseases as a result of its anti-inflammatory effects (8). Studying
immunosenescence by looking at master athletes, a population characterized by a longlife exposure to
exercise, could prove to be a valuable tool in understanding how exercise could improve life
expectancy. Preliminary results will be show about responses of Tregs and KLRG1 expression, a
marker associated a T cell activation and senescence (9), in naïve and memory Tregs to exercise in
master athletes.
References
[1] de Araujo AL. et al. Preventing or reversing immunosenescence: can exercise be an
immunotherapy? Immunotherapy. 2013, 5, 8.
[2] Simpson RJ. et al. Senescent T-lymphocytes are mobilised into the peripheral blood compartment
in young and older humans after exhaustive exercise. Brain Behav Immun. 2008, 22, 4.
[3] Andrews NP. et al. Telomeres and immunological diseases of aging. Gerontology. 2010, 56, 4.
[4] Yeh SH. et al. Regular tai chi chuan exercise enhances functional mobility and CD4CD25
regulatory T cells. Br J Sports Med. 2006, 40, 3.
[5] Maynard CL. et al. Diversity in the contribution of interleukin-10 to T-cell-mediated immune
regulation. Immunol Rev. 2008, 226.
[6] Gleeson M. et al. Respiratory infection risk in athletes: association with antigen-stimulated IL-10
production and salivary IgA secretion. Scand J Med Sci Sports. 2012, 22, 3.
[7] Wang J. et al. Effect of exercise training intensity on murine T-regulatory cells and vaccination
response. Scand J Med Sci Sports. 2012, 22, 5.
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Aveiro | 20th May 2015
3rd Workshop of Immunology
NOTES
Aveiro | 20th May 2015