O tratamento radioterápico continua aperfeiçoando-se, porém ainda pode estar associado com toxicidade pulmonar. Objetivo: Estudar os efeitos locais e sistêmicos provocados pela radioterapia conformacional torácica adjuvante em mulheres portadoras de câncer de mama, sem fatores de riscos prévios para desenvolvimento de alterações pulmonares. Por meio da tomografia computadorizada de alta resolução; identificar as possíveis alterações radiológicas no parênquima pulmonar. Se presentes, correlacioná-las com parâmetros obtidos da exploração funcional dos pulmões, com os efeitos sistêmicos, pela dosagem de mediadores inflamatórios, IL-1, IL-6 e TNF- , e suas repercussões sobre a qualidade de vida. Material e Métodos: Em 25 pacientes saudáveis foram coletadas amostras de sangue para serem utilizadas apenas como referência de normalidade da IL-1, IL-6 e TNF- . Em decorrência dos rígidos critérios de inclusão e exclusão estabelecidos, das 157 pacientes entrevistadas apenas 24 foram selecionadas para o estudo. A avaliação funcional pulmonar foi abrangente e constou de: medida dos volumes e capacidade dos pulmões, sendo o volume residual obtido pelo método de diluição do hélio em circuito fechado; estudo dos fluxos expiratórios máximos (curva fluxo x volume e curva volume x tempo) e medida da capacidade de difusão pulmonar pela técnica de respiração única do CO. A tomografia computadorizada de alta resolução (16 detectores) foi realizada no pré-planejamento da radioterapia conformacional do tórax, esta com dose total de 45-50 Gy em 25 frações. Para as dosagens de citocinas plasmáticas foram empregadas as técnicas de imunoabsorção enzimáticas (ELISA). Na avaliação da qualidade de vida foi aplicado o questionário Saint George´s Respiratory Questionnary traduzido e adaptado culturalmente ao Brasil. Todos estes procedimentos foram obtidos na fase pré e repedidos 3 meses após a radioterapia. Os resultados das duas fases foram comparados utilizando-se a versão exata do teste Wilcoxon e o teste de Correlação de Spearman, nível de significância p 0,05. Resultados: Entre parâmetros funcionais houve queda significativa apenas na difusão e no fluxo expiratório a 50% da capacidade vital forçada. Mesmo comportamento observou-se para citocina IL-6, já que os mesmos encontravam-se aumentados pré-RT. Não ocorreram mudanças significativas em nenhum dos domínios do questionário de qualidade de vida. As alterações tomográficas ocorreram 60,87% das pacientes na fase pós-radioterapia, em sua maioria de graus leves a moderados e não correlacionaram-se com as alterações observadas em outros parâmetros estudados. Conclusão: O aumento observado na IL-6 durante a fase pré-RT, parece ser um bom índice preditivo de alteração pulmonar. A capacidade de difusão foi a alteração mais evidente e parece ser o índice que melhor reflete as alterações pulmonares que afetam essas pacientes. Diante das discretas alterações tomográficas e funcionais observadas após a RT, é provável que para a redução observada na DLCO concorra uma combinação de fatores ao nível da membrana alvéolo-capilar. No conjunto, as alterações induzidas pela radioterapia conformacional nas pacientes estudadas foram de pequena monta, insuficientes para influenciar aspectos funcionais do pulmão e a qualidade de vida. / Radiotherapy continues to improve itself, but it may still be associated with pulmonary toxicity. Objective: To study the effects caused by local and systemic adjuvant thoracic conformal radiotherapy in women with breast cancer, without risk factors prior to the development of pulmonary alterations. Through high-resolution computed tomography, to identify the possible radiological alterations in the lung parenchyma. If positive identification occurs, correlates it with pulmonary functional parameters, its systemic effects, the dosage of inflammatory mediators IL-1, IL-6 and TNF-, and their impact on quality of life. Material and Methods: Blood samples were collected from 25 healthy patients to be used as a normal reference for IL-1, IL-6 and TNF- mediators. Due to the established strict inclusion and exclusion criteria, only 24 from the initial 157 interviewed patients were selected for this study. The evaluation of pulmonary function was comprehensive and included: Lung volume and capacity measurement through closed loop helium residual volume dilution method;Peak expiratory flow study (flow curve x volume and volume vs. time curve) and CO diffusing capacity measurement through single breath technique. High-resolution computerized tomography (16 detectors) was performed in the pre-planning phase of thoracic conformal radiotherapy where 45-50 Gy total dose was applied in 25 fractions. For cytokines plasma measurement, the enzymatic immunosorbent techniques (ELISA) were used. For quality of life assessment, the Brazil´s validated Saint George\'s Respiratory Questionnary was used. All these procedures were applied in the pre-phase and repeated three months later after radiotherapy sessions. The results of the two phases were compared using the exact version of the Wilcoxon test and Spearman correlation test with p 0.05 significance level. Results: Among the functional parameters, there were a significant decrease in dissemination and expiratory flow at 50% of forced vital capacity. The same behavior was observed for cytokine IL-6, since they were already high at pre-RT. There were no significant changes in any of the aspects of quality of life questionnaire. The tomographic alterations occurred in 60,87% of patients in the post radiotherapy phase, mostly having low to moderate degree and not correlated with the observed changes in other parameters. Conclusion: The IL-6 increase in the pre-RT phase appears to be a reasonable predictive index of pulmonary alterations. The diffusing capacity alterations were the most evident and seem to be the index that best reflects the pulmonary alterations that affect these patients. Given the discrete tomographic and functional abnormalities observed after RT, it is likely that for the observed reduction in DLCO compete a combination of factors occurring at alveolar-capillary membrane level. Overall, the conformal radiotherapy induced changes in the studied patients were not expressive, insufficient to influence the pulmonary functional aspects and the quality of life of the patients.

INTRODUÇÃO: O ozônio (O3) é encontrado na natureza e também pode ser produzido no corpo humano através da ativação de anticorpos. Seus efeitos anti-bactericidas são descritos na literatura, mas esses dados são controversos quanto a um potencial efeito benéfico da ozonioterapia no tratamento de certos tipos de infecção. OBJETIVO: Avaliar os efeitos da aplicação intraperitoneal (i.p.) de uma mistura gasosa de ozônio em um modelo de ligadura e punção de ceco (LPC) em ratos, através da dosagem de interleucinas (IL)-6, IL-10 e da quimiocina CINC-1 (cytokine-induced neutrophil chemoattractant), da lesão pulmonar aguda (LPA) e da análise das taxas de sobrevida. MÉTODO: Quatro grupos de ratos Wistar foram utilizados para análise de cada objetivo (CTR, LPC, LPC+O2 e LPC+O3). Os animais do grupo CTR foram submetidos somente a laparotomia. O grupo LPC foi submetido aos procedimentos de LPC. Os outros grupos foram submetidos à LPC e receberam injeção (i.p.) da mistura gasosa correspondente, administrada a cada 12 horas durante o período de observação. Os níveis séricos de IL-6, CINC-1 e IL-10 foram determinados por imuno- ensaio (enzyme linked immunosorbent assay- ELISA). A LPA foi avaliada através da histologia pulmonar e quantificada através do método do extravasamento pulmonar do Azul de Evans. Os animais da análise de sobrevivência foram observados por cinco dias. Os valores obtidos foramexpressos como médias ± erro-padrão da média (EP) ou medianas mais percentis 25 e 75(P25; P75), de acordo com a distribuição dos dados. Considerou-se significante p<0,05. RESULTADOS: Os ratos do grupo CTR exibiram os menores níveis de CINC-1 (p<0,01). O grupo LPC+O3 teve níveis menores de CINC-1 comparado a LPC+O2 e LPC (p<0,05). Os níveis de IL-10 do grupo CTR foram menores do que nos outros 3 grupos(p=0,02) . Não houve diferenças entre os outros 3 grupos (p=0,85). IL-6 foi significativamente menor para o grupo CTR (30,8± 4,8) quando comparado a todos os outros grupos (p<0,001). LPC+O3 e LPC+O2 exibiram níveis menores quando comparados ao grupo LPC (p<0,01). Não houve diferença entre os grupos LPC+O3 e LPC+O2 (p=0,54). O escore de histologia pulmonar foi menor para CTR (p=0,02). Os outros grupos não apresentaram diferenças significantes intergrupos (p=0,3). Os valores dos coeficientes de extravasamento pulmonar do Azul de Evans foram menores para LPC+O3 quando comparado aos grupos LPC+O2 e LPC (p=0,02), porém não houve diferença na comparação com CTR O grupo CTR teve o maior tempo de sobrevida (110±10h) comparado com os outros grupos, ou seja, LPC (57,3± 10,4h), LPC+O2 (71 ± 12,9h) e LPC+O3 (52,1 ± 8), os quais não apresentaram diferenças entre si quanto à sobrevida (p=0,4). CONCLUSÃO: No presente estudo experimental em ratos, a ozonioterapia teve um benefício potencial na modulação da resposta inflamatória e na LPA, mas não influenciou as taxas de sobrevida dos animais. / INTRODUCTION: Ozone (O3) is found in nature and also can be produced in the human body through activation of antibodies. Its antibacterial effect has been described in the literature, but these data are controversial regardi ng a benefic role of O3 therapy in the treatment of certain types of infection. OBJECTIVE: To evaluate the effects of intraperitoneal (i.p.) application of an O3 gas mixture in a rat model of cecal ligation and puncture (CLP), by analyzing interleukin (IL)-6, IL-10 and cytokine-induced neutrophil chemoattractant (CINC)-1 levels, acute lung injury (ALI) and survival rates. METHOD: Four animal groups were used (SHAM, CLP, CLP+O2 and CLP+O3). SHAM animals were submitted solely to laparotomy. CLP group was submitted to cecal ligation and puncture. The other groups were submitted to CLP and received injections (i.p.) of the corresponding gas mixture every 12 hours during the observation period. The serum concentrations of IL-6, CINC-1 and IL-10 were determined by the enzyme-linked immunosorbent assay (ELISA). ALI was evaluated with pulmonary histology and quantitated by means of the Evans blue dye (EBD) lung leakage method. For survival analysis, animals were observed for 5 days. Values were expressed as means ± SEM or medians (P25; P75), according to the data distribution. A p<0,05 was considered significant.RESULTS: SHAM rats had the lowest levels of CINC-1 compared to all other groups (p<0,01). CLP+O3 group had lower levels of CINC-1 compared to CLP+O2 and CLP (p<0,05). SHAM IL-10 levels were the lowest compared to the 3 other groups (p=0,02). There were no differences between the other 3 groups (p=0,85). IL-6 was significantly lower for SHAM compared to all groups (p<0,001). CLP+O3 and CLP+O2 had lower levels when compared to CLP (p<0,01). Comparison between groups CLP+O3 and CLP+O2 showed no significant difference (p=0,54). Pulmonary histology score was lower for SHAM (p=0,02). The other groups presented no statistical difference when compared to each other (p=0,3). EBD lung leakage values were lower to CLP+O3 compared to CLP+O2 and CLP (p=0,02). SHAM group had the longest survival time (110±10h) compared to all other groups (p=0,002). CLP (57,3± 10,4h), CLP+O2 (71 ± 12,9h) and CLP+O3 (52,1 ± 8h), which did not show difference on survival compared to each other (p=0,4). CONCLUSION: In this rat model of sepsis, ozone therapy had a potential benefit in the modulation of inflammatory response and ALI, but no improvement on survival rates was observed.

L'interleukine-6 (IL-6) exerce des propriétés biologiques multiples telles que l'activation des cellules immunocompétentes, l'activation de la réponse inflammatoire et l'hématopoïèse. Produite également par les cellules tumorales, l'IL-6 impacte la prolifération, la différenciation et la survie de ces dernières. L'IL-6 représente donc depuis plusieurs années une cible thérapeutique pertinente. Dans la première partie de ce travail, nous avons exploré une nouvelle piste potentielle pour bloquer l'activité biologique de l'IL-6, en utilisant un antagoniste naturel que notre équipe a identifié dans plusieurs lignées d'adénocarcinomes du rein, à savoir la molécule tronquée tIL-6. Suite à l'évaluation comparée de deux systèmes d'expression (E. coli versus CHO), nous avons retenu les cellules CHO comme source de production de fractions enrichies en tIL-6 par chromatographie de gel d'exclusion. Disposant d'un panel d'adénocarcinomes de rein (ACHN, Caki1, CLB CHA, CLB VER) et d'une lignée érythroleucémique (TF1), l'activité fonctionnelle de tIL-6 in vitro a été étudiée sur (1) la signalisation IL-6 induite, (2) la prolifération cellulaire IL-6 induite, la survie cellulaire et (4) la modulation de l'expression de protéines relevantes de l'apoptose. La molécule tIL-6 bloque la phosphorylation de la tyrosine Tyr705 de STAT3, qui est un des éléments clés de la voie de signalisation de l'IL-6. Nous rapportons également une autre observation nouvelle indiquant que tIL-6 exerce un effet pro-apoptotique sur certaines lignées RCC. Dans la seconde partie de notre étude, l'impact d'un Ac Mo anti IL-6 dans la réversion de la résistance aux cytotoxiques ou à la radiothérapie a été étudié. Nos résultats démontrent que la voie IL-6 ne constituerait pas un mécanisme majeur de résistance / Interleukin-6 (IL-6) plays numerous physiological roles including haematopoiesis, immune response and inflammation, but also plays a role in modulating cell growth, differentiation and survival of tumors cells. The first goal of the present study was to investigate on the potential role of the truncated protein IL-6 (tIL-6) encoded by the spliced IL-6 mRNA discovered in renal carcinoma cells (RCC). The R&D program was designed based on an industrial approach, aiming at reaching the decision stage to enter or not into preclinical development. Firstly two different expression systems were investigated (E. coli versus CHO cell line). The mammalian expression system was selected as the protein source since a recombinant glycosylated tIL-6 with a molecular weight similar to the predicted natural molecule was obtained from enriched fractions following size exclusion chromatography. Secondly by using a cell line panel including renal carcinoma cells (ACHN, Caki-1, CLB CHA, CLB-VER ) and an erythroleucemic cell line (TF1), in vitro tIL-6 functional activity were analyzed on (1) IL-6 induced signaling, (2) IL-6 induced cell proliferation, (3) on cell survival and also (4) on expression of specific set of proteins involved in apoptosis pathways. The truncated IL-6 was found inhibit IL-6 induced STAT3 Tyr705 and to induce apoptosis in some RCC cell lines which could be depending on IL-6 expression. Understanding more precisely the role of natural truncated IL-6 and its impact in cell tumour growth control will be a major issue in the development of innovative approach to antagonize directly or not IL6. The second goal of the present study was to investigate on reversing resistance of cancer cell lines to cytotoxics or ionizing radiations through the use of a monoclonal antibody directed against IL-6. Our data support the fact that IL-6 is not the preponderant actor of cell resistance to cytotoxics and ionizing radiations, which seems to be regulated by a complex network of proteins

Master of Science / Department of Clinical Sciences / James D. Lillich / Objective: To assess the in vitro ability of a novel lipopolysaccharide binding compound (EVK063) to inhibit cytokine production in lipopolysaccharide-stimulated equine peripheral blood mononuclear cells Animals: Eight healthy horses were sources for mononuclear cells. Procedures: Replicate aliquots (concentrated at 4-5 million cells/mL) were stimulated with S. typhimurium lipopolysaccharide (LPS) (100ng/mL), treated with graded concentrations of EVK063, (0.01µM, 0.1µM, 1µM, 10µM), Polymyxin B (PMB) (10µM) and incubated at 37°C for 6 hours. Media and cell samples were collected and stored at -80°C for evaluation of Tumor necrosis factor (TNF) using an equine specific ELISA and Interleukin-6 (IL6) via qRT-PCR. NanoDrop confirmed RNA quantity and primer sets designed for equine IL6 and the housekeeping gene 18s were used. EVK063 toxicity was evaluated with propidium iodide staining as determined by flow cytometry. Data was normalized, expressed as percent inhibition of cytokine up-regulation by LPS, and statistically evaluated by analysis of variance. Statistical significance was set at P ≤ 0.05. Results: Samples incubated in media with 0% serum demonstrated the following results: 0.01µM and 0.1µM EVK063 maintained >90% cellular viability yet failed to significantly inhibit TNF production or IL6 expression. The 1µM and 10µM EVK063 concentrations exhibited 25% and 70% cell death respectfully and therefore an interpretation as to their efficacy to inhibit TNF production or IL6 expression could not be made. Samples incubated in media with 10% serum demonstrated the following results: 0.01µM, 0.1µM and 1µM concentrations of EVK063 maintained >90% cellular viability yet failed to inhibit TNF production or IL6 expression. The 10µM EVK063 concentration exhibited 35% cell death and therefore an interpretation as to the efficacy to inhibit TNF production or IL6 expression could not be made. In a whole blood preparation, all samples evaluated maintained >90% cellular viability. The 10µM EVK063 significantly reduced TNF production and IL6 expression. Conclusion: This in vitro study confirms the ability of EVK063 to inhibit TNF production and IL6 expression in LPS stimulated equine mononuclear cells with comparable results to PMB.

B virus is an alphaherpesvirus, endemic to macaque monkeys, capable of deadly human zoonosis with an 80% mortality rate in untreated cases. The macaque monkey is widely used in biomedical research and the threat of B virus poses an occupational hazard to researchers, veterinarians, and animal handlers. B virus establishes a life-long latent infection in sensory neurons of the peripheral nervous system (PNS) in the natural host. In human infections, B virus readily transits to the central nervous system (CNS) and destroys brain tissues. Identifying immune correlates of B virus infection in the PNS of the natural host is critical in understanding viral lethality in the human host. The lack of an accurate animal model and restrictions on handling potentially infected nervous tissue previously limited studies of B virus infection in macaque ganglia. To address this barrier, a long-lived mixed neuron/glia cell culture model was established from macaque DRG explants using a novel methodology that relied on cellular migration from whole tissues. Utilizing this model, the hypothesis tested was that acute B virus infection of macaque ganglia triggers cellular defense networks to promote leukocyte recruitment and impact leukocyte activation. Chemokines were upregulated in B virus-infected cultures and infected cell media induced leukocyte chemotaxis. Leukocytes were less effectively activated by media from infected cells when compared to media from mock-infected cells. To identify factors responsible for this, focused microarrays were performed and cytokine profiles were quantified from B virus and mock-infected culture supernatants. IL-6 protein levels were significantly reduced in B virus infected cultures. This observation led to the hypothesis that IL-6 downregulation impairs leukocyte activation and, indeed, when IL-6 was added to B virus-infected culture supernatants to control levels, these cultures were far more effective at eliciting leukocyte activation when compared with mock-infected cultures. Collectively, these data support the hypothesis that acute B virus infection of macaque ganglia triggers cellular defense networks to promote leukocyte recruitment and impact leukocyte activation and identifies a potential viral mechanism to impair leukocyte functionality. Additionally, this work presents a novel methodology for establishing long-lived mixed neuron/glia cultures from postnatal/adult macaque DRGs.

Résumé : L’IL-7 et l’IL-15 sont des cytokines impliquées dans l’homéostasie des lymphocytes T CD8 naïfs et mémoires respectivement. Lors d’une réponse immunitaire, certaines cytokines pro-inflammatoires, comme l’IL-6 et l’IL-21, sont produites par les cellules du système immunitaire inné. Nous avons observé que certaines cytokines de ces deux groupes (homéostasie et pro-inflammatoires), peuvent avoir un effet synergique sur la fonction des lymphocytes T CD8. Spécifiquement, l’incubation des lymphocytes T CD8 naïfs avec l’IL-6 ou l’IL-21, en présence d’IL-7 ou d’IL-15 cause une forte prolifération qui est indépendante de l’antigène. De plus, la combinaison d’IL-15 avec l’IL-6 ou l’IL-21 entraîne une prolifération préférentielle des lymphocytes T mémoires, tandis que la combinaison avec l’IL-7 entraîne une prolifération des lymphocytes T naïfs. La stimulation des lymphocytes T CD8 avec l’IL-6 ou l’IL-21, en présence d’IL-7 ou d’IL-15, entraîne une augmentation de la phosphorylation en tyrosine de STAT5 ainsi qu’une augmentation de liaison à l’ADN. Nous avons étudié l’effet d’une pré-stimulation des cellules T CD8 naïves par les cytokines synergiques sur leur réponse subséquente à un antigène. Nous avons observé qu’une pré-stimulation avec l’IL-6 ou l’IL-21, en présence d’IL-7 ou d’IL-15, même pour une courte durée de 24 heures, augmente leur sensibilité aux antigènes, entraînant une robuste prolifération et une forte augmentation de cytotoxité spécifique à l’antigène gp33. Nous avons observé que les cytokines pro-inflammatoires en combinaison avec l’IL-7 induisent une augmentation accrue de la prolifération chez les lymphocytes T CD8 exprimant un TCR transgénique de forte affinité (P14), ainsi que les cellules exprimant un TCR de faible affinité (H-Y). De plus, la combinaison synergique de cytokines entraîne une forte expression du récepteur de l’IL-2R[gamma] (CD132), ainsi qu’une augmentation de la production d’IL-2 après stimulation antigénique. Une forte augmentation de l’expression de CD8 et de CD45, ainsi qu’une diminution drastique de l’expression de CD5 peut expliquer l’augmentation de l’avidité fonctionnelle du TCR suite à une stimulation avec les combinaisons de cytokines synergiques. La stimulation des lymphocytes T CD8 avec les combinaisons de cytokines, induit une augmentation de la phosphorylation de LAT ainsi qu‘AKT. Cependant, la stimulation subséquente du CD3 n’entraîne pas d’augmentation de la phosphorylation de LAT ainsi qu’AKT chez les lymphocytes T CD8 pré-stimulés avec les combinaisons de cytokines. Nous avons aussi observé que les lymphocytes T CD8 stimulés avec les combinaisons de cytokines augmentent l’expression de CD62L, ce qui peut favoriser leur migration vers les ganglions lymphatiques. En conclusion, la production de cytokines pro-inflammatoires (IL-6, IL-15, IL-21) par les cellules du système immunitaire inné lors d’une infection ou d’une inflammation, ainsi que la présence constitutive d’IL-7, peuvent stimuler la prolifération et l’activation des lymphocytes T CD8 de façon non spécifique à l’antigène. Cette stimulation entraîne une augmentation de l’avidité fonctionnelle de leur TCR causant ainsi une forte prolifération ainsi que l’acquisition de fonctions effectrices spécifiques. Cette liaison entre le système immunitaire inné et adaptatif, médiée par les cytokines pro-inflammatoires et les cytokines homéostatiques joue un rôle très important dans l’élimination des pathogènes ainsi que dans le développement de maladies auto-immunitaires. / Abstract : Homeostasis of naive and memory CD8[superscript +] T lymphocytes is dependent on two cytokines IL-7 and IL-15, respectively. During an immune response to an infection, cells of the innate immune system produce several pro-inflammatory cytokines. We have observed that these two groups of cytokines, namely proinflammatory and homeostatic, can have a synergistic effect on CD8 T lymphocytes. Specifically, incubation of naive CD8 T cells with IL-6 or IL-21 in the presence of IL-7 or IL-15 induced strong proliferation in an antigen independent manner. While the combination of IL-6 or IL-21 with IL-15 induced strong proliferation of memory CD8 T cells, naïve CD8 T cells responded better to the combination with IL-7. These stimulatory cytokine combinations elicited strong STAT5 phosphorylation and it’s binding to DNA in CD8 T cells. We investigated the effect of priming CD8 T cells with the synergistic combination of IL-6 or IL-21 and IL-7 on their subsequent response to antigen. We observed that cytokine priming for only 24 hours enhanced their sensitivity to antigen, resulting in strong proliferation, effectors functions and cytotoxicity. These effects were observed with CD8 T cells expressing transgenic TCR with strong (P14) or weak (H-Y) affinity towards cognate peptide antigens. Priming CD8 T cells with the synergistic combination of cytokines increased the expression of IL-2 receptor gamma (CD132) and augmented the production of IL-2 when stimulated with antigen. These cells also expressed elevated levels of CD8 and CD45, as well as down modulate CD5, and these events may underlie the increased TCR avidity. Stimulation of CD8 T cells with the synergistic combination of cytokines induced phosphorylation of LAT and AKT. However, subsequent TCR stimulation did not further increase these phosphorylation events. We have observed that C D8 T cells primed with the synergistic combinations of cytokines up regulated CD62L, which could promote their migration through lymph nodes. In conclusion, inflammatory cytokines such as (IL-6, IL-15, IL-21) secreted by cells of the innate immune system during an infection or non-infectious inflammation, and basal levels of the homeostatic cytokine IL-7 can act in synergy with inflammatory cytokines to activate CD8 T lymphocytes in an antigen independent manner. This stimulation also results in an increase in the functional avidity of their TCR, as indicated by strong antigen responsiveness with increased proliferation and display of effectors functions. This connection between the innate and adaptive system mediated by inflammatory cytokines may play an important role in pathogen clearance and possibly in the development of autoimmune diseases.

No description available.

The IL-6 system is key in the development of chronic low-grade inflammation. It is known to be upregulated in response to acute exercise and lowered at rest after exercise training. IL-6 has both anti- and pro-inflammatory properties and moderation of this cytokine could alleviate chronic low-grade inflammation which is associated with obesity and Type 2 diabetes mellitus (T2DM). This thesis investigated the interplay between inflammation, glycaemic control and high intensity intermittent training (HIIT) - an exercise regimen that has been shown to yield many health benefits. There was a greater increase in IL-6 after an acute bout of HIIT than continuous moderate intensity exercise, where external work was matched (Chapter 4). Although sIL-6R and the IL-6/sIL-6R complex were both significantly increased after acute exercise there were no differences between HIIT and moderate intensity exercise. In response to 2 weeks HIIT there was a significant reduction in IL-6 and increase in IL-6R in adipose tissue in overweight and obese males (Chapter 5). It was also determined that IL-6R present in adipose tissue is at least partly composed of the membrane-bound IL-6R isoform (Chapter 6). Reductions in circulating sIL-6R, the IL-6/sIL-6R complex, MCP-1 and adiponectin, as well as a decrease in waist circumference and increase in peak oxygen uptake during exercise were also induced after 2 weeks HIIT (Chapter 5). Young adults with T2DM (< 40 y) displayed elevated levels of inflammatory proteins in comparison to lean controls, however there were no significant differences in comparison to obese controls (Chapter 7). In conclusion, the findings of this thesis demonstrate that acute and repeated bouts of HIIT have positive effects on the inflammatory profile in the circulation and adipose tissue, particularly in relation to the IL-6 system. It should be determined if HIIT is an achievable mode of exercise for patient populations, including T2DM patients, in order to downregulate the inflammatory profile.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

O carcinoma espinocelular de boca (CEB) é a neoplasia maligna mais comum da cavidade oral, correspondendo à aproximadamente 94% dos casos dessa região. Apesar dos diversos estudos moleculares e celulares do CEB, a taxa de sobrevida dos pacientes é de aproximadamente 50%, devido principalmente ao tamanho do tumor, metástase em linfonodos regionais, grau de diferenciação das células e sítio anatômico. O microambiente tumoral do CEB, é extremamente complexo e diversificado, tendo como característica principal um estado inflamatório crônico imunossupressivo. Este microambiente é sustentado pela liberação de diferentes citocinas inflamatórias, como IL-6, TNF- - atividades exercidas tanto pelas células tumorais quanto pelas estromais. Dentre essas atividades, tem sido relatado na literatura que as citocinas inflamatórias são capazes de aumentar a migração e a capacidade de invasão das células tumorais. Entre as células estromais, os macrófagos são as mais abundantes e participam da manutenção do microambiente tumoral. De acordo com o estímulo, podem ser polarizados M1, com papel pró-inflamatório e antitumoral, e M2, com papel anti-inflamatório e pró-tumoral. O objetivo desta tese foi compreender o papel dos macrófagos no prognóstico de CEB e das citocinas inflamatórias IL-6, TNF- - linhagens celulares de CEB. Para verificar o papel dos macrófagos no prognóstico, foi realizada uma revisão sistemática na qual foram incluídos apenas os estudos que utilizavam amostra de pacientes com CEB e avaliavam o prognóstico com marcadores para macrófagos. Foi observado que maiores concentrações de macrófagos CD68+ e CD163+ estavam relacionados com pior prognóstico de pacientes com CEB, embora não tenha sido possível concluir qual região tumoral a presença destas células seja mais importante 7 para o desfecho. Para analisar o papel das citocinas inflamatórias IL-6, TNFILensaios in vitro utilizando duas linhagens celulares, SCC25 e Cal27, em condições promotoras de migração sob a influência dessas citocinas. Foi observado que a citocina IL-6 foi capaz de aumentar a velocidade de migração e a direcionalidade tanto da SCC25 quanto da Cal 27 e que esta melhora na capacidade migratória ocorreu através de um crosstalk entre a via de sinalização relacionada a IL6 (STAT3) e a via reguladora de migração celular, Rho GTPase Rac1. Estes dados reforçam o papel do microambiente tumoral no processo de progressão tumoral e sugerem potenciais alvos terapêuticos como a modulação do perfil da população de macrófagos e o papel de interleucinas no controle de invasão tecidual e metástase. / Oral squamous cell carcinoma (OSCC) is the most common malignant neoplasm of the oral cavity, corresponding to approximately 94% of the cases in this region. Despite the diverse molecular and cellular studies of OSCC, the patient survival rate is approximately 50%, mainly due to tumor size, regional lymph node metastasis, cell differentiation and anatomic site. The OSCC tumor microenvironment is extremely complex and diverse, with the main characteristic being an immunosuppressive chronic inflammatory state. This microenvironment is supported by the release of different inflammatory cytokines, such as IL-6, TNF- - and enhance the activities of both tumor and stromal cells. Among these activities, it has been reported in the literature that inflammatory cytokines are capable of increasing migration and invasiveness of tumor cells. Among stromal cells, macrophages are the most abundant and participate in the maintenance of the tumor microenvironment. According to the stimulus, macrophages can be polarized in M1, with pro-inflammatory and anti-tumoral role, and M2, with antiinflammatory and pro-tumoral role. Thus, the aim of this thesis was to evaluate the role of macrophages in the prognosis of OSCC and the influence of inflammatory cytokines IL-6, TNF- - OSCC cell lines. To assess the role of macrophages in the prognosis, a systematic review was conducted in which only studies using a sample of OSCC patients were evaluated and the prognosis was evaluated with macrophage markers. It was observed that higher concentrations of CD68 + and CD163 + macrophages were related to worse prognosis in patients with OSCC, although it was not possible to conclude which tumor region the presence of these cells is more important for the outcome. In order to analyze the role of the inflammatory cytokines IL-6, TNF- - atory 9 behavior of OSCC cells, in vitro assays using two cell lines, SCC25 and Cal27, were performed in migration-promoting conditions under the influence of these cytokines. It was observed that IL-6 was able to increase the speed migration and directionality of both SCC25 and Cal 27 and that this improvement in migratory capacity occurred through a crosstalk between the IL6-related signaling pathway (STAT3) and cell migration-related pathway, RhoGTPase Rac1. These data reinforce the role of the tumor microenvironment in the tumor progression process and suggest potential therapeutic targets such as the modulation of the profile of the macrophages population and the role of interleukins in the control of tissue invasion and metastasis.