Depuis les 15 dernières années, de nombreuses études ont mis en évidence le rôle majeur de l’architecture nucléaire dans la régulation de l’expression des gènes et ceci dans une grande diversité de processus biologiques. Bien que la réponse immunitaire soit un de ces processus, peu de données existent sur l’organisation spatiale du génome dans les cellules immunitaires et son impact sur la régulation des gènes dans le contexte d’une réponse à l’infection bactérienne. En utilisant le porc comme organisme modèle, nous avons concentré notre étude sur les macrophages dérivés de monocytes, premières lignes de défense contre les pathogènes. Les cellules immunitaires étant les cibles des mycotoxines, nous nous sommes également intéressés aux effets de la toxine T-2 sur les macrophages et leur réponse induite par les récepteurs TLR. Un effet cytotoxique de la T-2 à une dose de 10 nM, ainsi que des effets inhibiteurs sur certaines réponses liées aux récepteurs TLR ont été mis en évidence. Nous avons ensuite examiné si les changements dans l'expression des gènes dus à l'activation impliquent un repositionnement dans l'espace nucléaire. Une analyse du transcriptome a permis d’identifier les gènes différentiellement exprimés dans les macrophages activés par le mélange LPS-IFNγ et de mettre en évidence des réseaux de gènes impliqués lors de l’activation. Les 4 gènes les plus sur-exprimés (IL1β, IL8, CXCL10 et TNFα) et les 4 gènes les plus sous-exprimés (VIM, LGALS3, TUBA3 et IGF2) ont été sélectionnés pour analyser leur comportement dans l'espace nucléaire au cours de l’activation des macrophages en utilisant la technique FISH 3D. Parmi les 4 gènes sur-exprimés, 3 présentent des modifications de leur position durant le processus d'activation alors que les 4 gènes sous-exprimés ne montent pas de variation de leur position. L'analyse de la position des gènes par rapport à leurs territoires chromosomiques (TC) a été étendue à un second type de cellules immunitaires : les neutrophiles. Des résultats similaires ont été obtenus. Les analyses ont été ensuite complétées par l'étude des 4 gènes sur-exprimés dans un type cellulaire indépendant de la réponse immunitaire (fibroblastes). Nos données suggèrent que les relocalisations dans l'espace nucléaire des gènes différentiellement exprimés dans les cellules immunitaires activées sont gène spécifique, type cellulaire spécifique et concernent essentiellement ceux qui sont sur-exprimés. / For the past 15 years, numerous studies have highlighted the role of nuclear architecture in regulating gene expression in a variety of biological processes. Although the immune response is one of these processes, few data exist on the spatial organization of the genome in immune cells and its impact on gene regulation in the context of a response to bacterial infection. Using pigs as a model organism, we focused our study on monocytederived macrophages, the first lines of defence against pathogens. As immune cells are the targets of mycotoxins, we also studied the effects of T-2 toxin on macrophages and on their responses induced by TLRs. A cytotoxic effect of T-2 at a dose of 10 nM, and an inhibitory effect on some responses related to TLRs have been demonstrated. We then examined whether changes in gene expression due to activation involve a repositioning within the nuclear space. A transcriptome analysis allowed us to identify differentially expressed genes in macrophages activated by LPS-IFNγ and to analyze gene networks involved during activation. The top 4 up-regulated genes (IL1 beta, IL-8, CXCL10 and TNF) and the top 4 down-regulated ones (VIM LGALS3, TUBA3 and IGF2) have been selected to analyze their behaviour in nuclear space during macrophages activation using 3D FISH technique. Among the 4 up-regulated genes, 3 show changes in their position during the activation process while the 4 down-regulated ones did not reposition. The analysis of gene behaviours towards their chromosome territories (CT) was extended to neutrophils and similar results were obtained. The analyses were then completed by the study of the 4 up-regulated genes in a non immune cell type (fibroblast). Our data suggest that relocation in the nuclear space of genes differentially expressed in activated immune cells is gene and cell type specific and mostly concerns those that are up-regulated.

Mycobacterium tuberculosis (Mtb) is the pathogen causing tuberculosis (TB), a disease most often affecting the lung. 1.5 million people die annually due to TB, mainly in low-income countries. Usually considered a disease of the poor, also developed nations recently put TB back on their agenda, fueled by the HIV epidemic and the global emergence of drug-resistant Mtb strains. HIV-coinfection is a predisposing factor for TB, and infection with multi-drug resistant and extremely drug resistant strains significantly impedes and lengthens antibiotic treatment, and increases fatality. Mtb is transmitted from a sick individual via coughing, and resident macrophages are the first cells to encounter the bacterium upon inhalation. These cells phagocytose intruders and subject them to a range of destructive mechanisms, aiming at killing pathogens and protecting the host. Mtb, however, has evolved to cope with host pressures, and has developed mechanisms to submerge macrophage defenses. Among these, inhibition of phagosomal maturation and adaptation to the intracellular environment are important features. Mtb profoundly alters its phenotype inside host cells, characterized by altered metabolism and slower growth. These adaptations contribute to the ability of Mtb to remain dormant inside a host during latent TB infection, a state that can last for decades. According to recent estimates, one third of the world’s population is latently infected with Mtb, which represents a huge reservoir for active TB disease. Mtb is also intrinsically tolerant to many antibiotics, and adaptation to host pressures enhances tolerance to first-line TB drugs. Therefore, TB antibiotic therapy takes 6 to 9 months, and current treatment regimens involve a combination of several antibiotics. Patient noncompliance due to therapeutic side effects as well as insufficient penetration of drugs into TB lesions are reasons for treatment failure and can lead to the rise of drug-resistant populations. In view of the global spread of drug-resistant strains, new antibiotics and treatment strategies are urgently needed. In this thesis, we studied the interplay of the primary host cell of Mtb, human macrophages, and different Mtb phenotypes. A low-burden infection resulted in restriction of Mtb replication via phagolysosomal effectors and the maintenance of an inactive Mtb phenotype reminiscent of dormant bacteria. Macrophages remained viable for up to 14 days, and profiling of secreted cytokines mirrored a silent infection. On the contrary, higher bacterial numbers inside macrophages could not be controlled by phagolysosomal functions, and intracellular Mtb shifted their phenotype towards active replication. Although slowed mycobacterial replication is believed to render Mtb tolerant to antibiotics, we did not observe such an effect. Mtb-induced macrophage cell death is dependent on ESAT6, a small mycobacterial virulence factor involved in host cell necrosis and the spread of the pathogen. Although well-studied, the fate of ESAT6 inside infected macrophages has been enigmatic. Cultivation of Mtb is commonly carried out in broth containing detergent to avoid aggregation of bacilli due to their waxy cell wall. Altering cultivation conditions revealed the presence of a mycobacterial capsule, and ESAT6 situated on the mycobacterial surface. Infection of macrophages with this encapsulated Mtb phenotype resulted in rapid ESAT6-dependent host cell death, and ESAT6 staining was lost as bacilli were ingested by macrophages. These observations could reflect the earlier reported integration of ESAT6 into membranes followed by membrane rupture and host cell death. In conclusion, the work presented in this thesis shows that the phenotype of Mtb has a significant impact on the struggle between the pathogen and human macrophages. Taking the bacterial phenotype into account can lead to the development of drugs active against altered bacterial populations that are not targeted by conventional antibiotics. Furthermore, deeper knowledge on Mtb virulence factors can inform the development of virulence blockers, a new class of antibiotics with great therapeutic potential.

A leishmaniose é um termo global para doenças causadas por parasitos do gênero Leishmania, sendo a Leishmaniose Visceral (LV) a forma mais grave da doença. No Brasil é causada pelo parasita Leishmania (Leishmania) infantum chagasi. Para garantir a sua sobrevivência, alguns parasitas são capazes de manipular respostas de defesa das células do sistema imune. Recentes estudos demonstraram a participação da proteína imunomodulatória CD200 durante o processo de infecção de L. (L.) amazonenses. O presente estudo teve como objetivo investigar se os parasitos L. (L.) infantum chagasi são capazes de induzir a expressão da proteína CD200 durante o processo infeccioso. Em ensaios de infecção ex vivo, não foi observado proliferação de parasitas intracelulares. Apesar disso, L. (L.) infantum chagasi foi capaz de induzir a expressão do gene CD200. De maneira interessante, diferente de infecções por L. (L.) amazonenses, a indução de CD200 nessas células foi observada em tempos mais tardios de infecção. Ensaios de imunoprecipitação e Western blot indicaram a síntese da proteína, que atingiu os seus maiores níveis a 120 horas pós-infecção. A presença de CD200 sugere o envolvimento dessa molécula em tempos mais tardios de infecção por L. (L.) infantum chagasi. / Leishmaniasis is a global term for diseases caused by parasites of the genus Leishmania, and Visceral Leishmaniasis (VL) are the most severe form of the disease. In Brazil is caused by the parasite Leishmania (Leishmania) infantum chagasi. To ensure their survival, some parasites can handle defensive responses of the cells of the immune system. Recent studies have demonstrated the participation of immunomodulatory protein CD200 during the infection process of L. (L.) amazonenses. This study aimed to investigate whether the parasites L. (L.) infantum chagasi are capable of inducing the expression of CD200 protein during the infectious process. In trials of ex vivo infection, there was no proliferation of intracellular parasites. Nevertheless, L. (L.) infantum chagasi was able to induce the expression of CD200 gene. Interestingly, unlike infection by L. (L.) amazonenses, CD200 induction of these cells was observed at later times in infection. Immunoprecipitation assays and Western blot indicated protein synthesis, which reached their highest levels at 120 hours post-infection. The presence of CD200 suggests the involvement of this molecule at later times of infection with L. (L.) infantum chagasi.

O diabetes mellitus tipo I (DMI) é uma doença crônica autoimune caracterizada por hiperglicemia devido à destruição das células beta pancreáticas produtoras de insulina. Ao final de 30 dias da indução do diabetes por estreptozotocina, os macrófagos peritoneais residentes dos animais diabéticos apresentaram aumento de RNAm de citocinas e quimiocinas inflamatórias, secreção de óxido nítrico, expressão de NLRP3, iNOS e PARP1 e da atividade da via glicolítica. Perfil pró-inflamatório também foi observado em macrófagos peritoneais de animais NOD (non-obese diabetic). Camundongos diabéticos deficientes em NLRP3 (NLRP3 KO) apresentaram diminuição na expressão de iNOS, PARP1 e na produção de NO em relação aos macrófagos dos animais diabéticos selvagens. O estado diabético tipo I influenciou o perfil dos macrófagos peritoneais residentes, causando aumento na produção de NO, via NLRP3-PARP1-iNOS, expressão de citocinas pró-inflamatórias, receptores de quimiocinas e da atividade glicolítica. O tratamento com DHA (ômega-3) ex-vivo reverteu este perfil e atenuou o quadro pró-inflamatório por diminuição da produção de NO e da expressão de citocinas pró-inflamatórias. / Type I diabetes mellitus (DMI) is a chronic autoimmune disease characterized by hyperglycemia due to the destruction of insulin-producing pancreatic beta cells. At the end of 30 days after type I diabetes induced by streptozotocin, macrophages from diabetic animals had increased expressions of inflammatory cytokines and chemokines, secretion of nitric oxide, expression of NLRP3, iNOS and PARP1, and glycolytic activity compared to the cells from control animals. Proinflammatory features was also observed in peritoneal macrophages of NOD (non-obese diabetic) animals. Macrophages from NLRP3 deficient diabetic mice (NLRP3 KO) had decreased expression of iNOS, PARP1 and of NO production when compared to cells from wild type animals. The type I diabetic state led to a proinflammatory feature in resident peritoneal macrophages by increasing NO production, via the NLRP3-PARP1-iNOS pathway, expressions of proinflammatory cytokines, chemokine receptors and glycolytic activity. In contrast, ex-vivo treatment with DHA (omega-3) reversed this profile and attenuated the proinflammatory state by reducing NO production and expression of proinflammatory cytokines.

O envelhecimento é marcado por inúmeras alterações fisiológicas, dentre as quais encontramos um aumento da gordura corporal e alterações na resposta do sistema imunológico. Muito se sabe sobre a intrínseca ligação entre o aumento de gordura e as doenças de risco e fica a questão à cerca de sua influência sobre as respostas do sistema imunológico, já que estes dois fatores encontram-se alterados no envelhecimento. Portanto, o propósito deste estudo foi investigar a interação entre a quantidade de gordura corporal e as células do sistema imunológico de ratos envelhecidos. Para isso, num primeiro estudo, foram utilizados ratos ADULTOS e ENVELHECIDOS para a caracterização das alterações encontradas no processo de envelhecimento. Numa segunda etapa, os animais ENVELHECIDOS foram submetidos a dois protocolos de emagrecimento, a restrição calórica a 50%(RC) e o exercício aeróbio (EX) (natação), durante o período de quatro a seis semanas. Assim, foi possível avaliar alguns parâmetros do metabolismo de macrófagos e linfócitos em ratos envelhecidos com diferentes quantidades de gordura corporal. No primeiro estudo, os resultados comprovaram o aumento da gordura bem como algumas alterações no sistema imunológico de ratos envelhecidos, principalmente na capacidade funcional de macrófagos. Quanto ao segundo estudo, os resultados demonstram que ambos os protocolos foram eficientes em diminuir o peso corporal, bem como a gordura, sendo que para a RC os valores alcançados foram mais evidentes. Com relação ao sistema imunológico houve um aumento da resposta fagocitária e de produção de H2O2 por macrófagos e uma alteração da capacidade proliferativa de linfócitos, em ambos os protocolos. / Ageing is marked by several kinds of physiological changes, among then we find an increase of fat mass and a decline in several aspects of the immune system. It is well understood the intrinsical connexion between the increase in body fat and the risk of related diseases and the one question that remains is about the influence over the immune system response, since these two factors are altered on the ageing process. The purpose of this study was to investigate the relationship among fat content and immune cells of the aging rats. For this, in one first study, we used ADULTS and AGEING rats to determine the changes found in the ageing process. In the second time, the AGEING rats were submitted to two protocols for weight loss (slimming), 50% of caloric restriction (CR) and aerobic exercise (swimming) (EX), for four or six weeks. In this way, we can study some parameters of macrophages and lymphocytes metabolism in ageing rats with different body fat content. In the first study, the results confirmed the fat increase as well as some changes in the immune system of the ageing rats, especially in the macrophages functional capabilities. In the second study, the results showed that both protocols decreased body fat and weight, with the best results happening with CR protocol. The immune system had an increase on the phagocytic response and H2O2 production for macrophages and alterations of the proliferative capacities of the lymphocytes, in both protocols.

Introdução Os fumantes inalam grandes quantidades de partículas de carbono, o que pode contribuir para efeitos adversos pulmonares e sistêmicos. É sabido que os macrófagos alveolares (MA) desempenham um papel extremamente importante no reconhecimento e processamento de qualquer material estranho inalado e são as células predominantes que processam e removem partículas inaladas. Existe também a deposição superficial a longo prazo do carbono observado nos pulmões de fumantes em autópsias. Atualmente, a distribuição e retenção de partículas de fumo derivadas de cigarros quando a pessoa também está exposta a níveis elevados de poluição do ar ainda não é clara. Portanto, procurou-se avaliar a carga de carbono nos MA e a deposição de superfície pulmonar em uma população exposta a alta poluição atmosférica (São Paulo), tanto em fumantes como não-fumantes. Métodos Uma coorte de 72 sujeitos post mortem foi obtida do Serviço de Verificação de Óbitos da Capital da Universidade de São Paulo (SVOC). As imagens das superfícies pulmonares foram obtidas sob condições padrão e pequenos fragmentos de tecido pulmonar foram coletados para análise de macrófagos usando a técnica de esfregaço. A superfície total de negro de carbono foi analisada utilizando o programa Imagem J (National Institute of Health, MD, EUA), teste cego ao fumo. A absorção interna de carbono nos MA foi medida utilizando o programa Image Pro Plus (The Proven Solution, Media Cybernetics Inc., EUA). A aprovação ética foi obtida. A média de negro de carbono de macrófagos tanto em fumantes como em não-fumantes foi analisada utilizando teste de Mann Whitney e expressa como intervalo interquartil (IQR). Resultados Os fumantes têm um nível significativamente mais elevado de negro de carbono nos macrófagos alveolares (103.4 (IQR 29.44 to 226.3) vs. 9.27 (IQR 3.1 to 85.13) um2, P < 0.001)103.4um2. Não houve diferença significativa entre a área média de deposição superficial de carbono nos pulmões de fumantes e não fumantes de 6, 74 cm2 (IQR 3, 47 a 10, 02) versus 5, 20 cm2 (IQR 2, 29 a 7, 54) P=NS. Conclusão O teor de carbono nos MA é claramente muito maior nos fumantes do que os nãofumantes. No entanto, a análise da superfície pulmonar não mostrou diferença significativa. Isso pode indicar que, em uma área de alta poluição do ar, o principal fator que contribui para a deposição de carbono no pulmão a longo prazo é a exposição à poluição com efeitos limitados da exposição à fumaça de cigarro. O preto de carbono nos MA ainda aparece significativamente influenciado pela exposição à fumaça de cigarro / Rationale Smokers inhale large amounts of carbonaceous particulate matter, which may contribute to pulmonary and systemic adverse effects. It is clear that alveolar macrophages (AM) play a critically important role in the recognition and processing of any inhaled foreign material and are the predominant cells that process and remove inhaled particulate matter from the lung. There is also long-term surface deposition of carbon seen in the lungs of smokers at post-mortem. At present the distribution and retention of cigarette smoke-derived particulate matter when the person is also exposed to high levels of background air pollution is unclear. Therefore we sought to assess both AM carbon loading and lung surface deposition in a population exposed to high background air pollution (São Paulo) in both smokers and non-smokers. Methods A cohort of 72 post-mortem subjects was obtained from São Paulo Autopsy Centre (SVOC). Images of lung surfaces were obtained under standard conditions and small fragments of lung tissue were collected for macrophage analysis using smear technique. The total surface black carbon was analysed using Image J (National Institute of Health, MD, USA), blinded to smoking status. Internal AM carbon uptake was measured using Image Pro Plus (The Proven Solution, Media Cybernetics Inc., USA). Ethical approval was obtained. Mean macrophage black carbon in both smokers and non-smokers was analysed using Mann Whitney and expressed as median (IQR). Results Smokers have a significantly higher level of mean macrophage black carbon (103.4 (IQR 29.44 to 226.3) vs. 9.27 (IQR 3.1 to 85.13) um2, P < 0.001)103.4um2. There was no significant difference between the mean area of surface deposition of carbon in the lungs of smokers and non-smokers 6.74 cm2 (IQR 3.47 to 10.02) versus 5.20cm2 (IQR 2.29 to 7.54) P=NS. Conclusion AM carbon content is clearly much higher in the smokers than the non-smokers. However the lung surface analysis showed no significant difference. This could indicate that in an area of high air pollution the main contributing factor to long term lung carbon deposition is pollution exposure with limited effects from cigarette smoke exposure. AM black carbon still appears significantly influenced by cigarette smoke exposure

As doenças cardiovasculares são a principal causa de mortalidade no mundo. A aterosclerose é a base fisiopatológica dessas doenças, sendo definida como um processo crônico-inflamatório multifatorial, resultando da interação de diferentes células como linfócitos, macrófagos, células endoteliais e células musculares lisas na parede arterial. A lipoproteína de baixa densidade eletronegativa [LDL(-)], uma subfração modificada da LDL nativa, desempenha um papel-chave na aterosclerose, uma vez que as modificações sofridas por esta partícula são capazes de induzir o acúmulo de ésteres de colesterol em macrófagos e a subsequente formação de células espumosas. O sistema imunológico é crucial no processo aterogênico e estratégias terapêuticas direcionadas à imunoregulação deste processo têm sido utilizadas como novas alternativas tanto na prevenção do desenvolvimento quanto da progressão desta doença. Dentre essas estratégias, destaca-se o uso de fragmentos de anticorpos como o scFv (do inglês, single chain fragment variable), que podem ainda estar conjugados a nanopartículas com o intuito de aumentar sua eficiência de ação no organismo. Diante do papel da LDL(-) na aterosclerose, este projeto objetivou avaliar os efeitos in vitro e in vivo de um sistema nanoestruturado contendo fragmentos scFv anti-LDL(-) derivatizados na superfície de nanocápsulas sobre macrófagos murinos e humanos primários e em camundongos knockout para o gene do receptor da LDL (Ldlr-/-) no desenvolvimento e na progressão dessa doença. Demonstrou-se que o tratamento de macrófagos com a formulação scFv anti-LDL(-)-MCMN-Zn diminuiu de forma significativa a captação de LDL(-), assim como a expressão de IL-1&#946; (mRNA e proteína) e MCP-1 (mRNA). Foi demonstrada a internalização da nanoformulação pelos macrófagos via diferentes mecanismos de endocitose, demonstrando seu potencial uso como carreador de fármacos. In vivo, a nanoformulação diminuiu de forma significativa a área da lesão aterosclerótica em camundongos Ldlr-/- submetidos à avaliação pela técnica de tomografia por emissão de pósitrons (do inglês, PET), utilizando o radiotraçador 18F-FDG (18F-desoxiglicose), associada à tomografia computadorizada (CT) com agente de contraste iodado, além da análise morfométrica das lesões no arco aórtico. O conjunto dos resultados obtidos evidenciou a ação ateroprotetora da formulação scFv anti-LDL(-)-MCMN-Zn, reforçando seu potencial como estratégia terapêutica na aterosclerose. / Cardiovascular diseases are the leading cause of mortality worldwide. Atherosclerosis is the pathophysiological basis of these diseases, defined as a chronic inflammatory multifactorial process, resulting from the interaction of several cells such as lymphocytes macrophages, endothelial cells and smooth muscle cells within the arterial wall. The electronegative low-density lipoprotein [LDL(-)], a modified subfraction of native LDL, plays a key role in atherosclerosis, since its modifications are capable of inducing the accumulation of cholesteryl esters in macrophages and the subsequent foam cells formation. The immune system is crucial in atherogenic process and therapeutic strategies directed to the immunoregulation of this process have been used as a new alternative in the prevention of the development as well as the progression of this disease. Among these strategies, it is the use of antibody fragments such as scFv (single chain fragment variable), which may be also conjugated to nanoparticles in order to increase their efficiency in the body. Given the role of LDL(-) in atherosclerosis, the aim of this project was to evaluate the in vitro and in vivo effects of a nanostructured system containing scFv anti-LDL(-) fragments derivatized on the surface of nanocapsules on murine and human primary macrophages and in the development and progression of the disease in LDL receptor knockout mice (Ldlr-/-). It was demonstrated that the treatment of macrophages with scFv anti-LDL(-)-MCMN-Zn formulation significantly decreases the uptake of LDL(-) and the expression IL-1&#946; (mRNA and protein) and MCP-1 (mRNA). Moreover, the internalization of the nanoformulation by macrophages through different endocytosis mechanisms was shown, demonstrating its potential use as a nanocarrier. In vivo, the nanoformulation decreased the area of atherosclerotic lesions in Ldlr-/- mice evaluated by positron emission tomography with 18F-FDG associated with computed tomography with iodinated contrast agent (PET/CT), besides the lesion morphometric analysis at the aortic arch Thus, these data provide evidence of the atheroprotection action of the ateroprotection action of the scFv anti-LDL(-)-MCMN-Zn formulation, suggesting its promising use as a therapeutic strategy for atherosclerosis.

A cirrose é uma doença cosmopolita que causa milhares de mortes a cada ano e encontra-se em estado de atenção pelos sistemas de saúde governamentais. Em auxílio à sua compreensão, utiliza-se o modelo de indução de cirrose pela tioacetamida (TAA) em ratas. Porém este modelo tem carência de informações sobre a presença de células inflamatórias no fígado, as quais desempenham ação destrutiva e estimulam o desenvolvimento da cirrose. Propusemos nesta pesquisa a detecção de macrófagos e células dendríticas neste modelo nas primeiras sete semanas com o uso de tioacetamida na dose de 200mg/kg em aplicações intraperitoneais, correlacionando-as com a apoptose de hepatócitos, a presença de células MCP-1 positivas e o desenvolvimento da fibrose em fígado de ratas. Com isto, esperou-se encontrar o aumento contínuo de macrófagos teciduais e exsudativos, juntamente com o aumento de células dendríticas, atraídas pela secreção de MCP-1 pelas células estreladas hepáticas. Esperou-se também a detecção de hepatócitos em apoptose e consequente deposição de fibras colágenas pelas células estreladas hepáticas. A ação tóxica da tioacetamida foi observada pela necrose de sacabocado e reação tecidual com proliferação de ductos biliares. A tioacetamida foi eficiente em desencadear a presença de macrófagos e hepatócitos em apoptose nas primeiras semanas, contudo foi ineficaz em manter o aumento destes eventos de forma constante até o final da sétima semana. Além disso, o aumento do tecido colágeno não foi significante, indicando indiretamente a presença de macrófagos e a lesão de hepatócitos não foi suficientemente intensa para desencadear a ação máxima das células estreladas hepáticas durante as sete semanas de tratamento. A utilização de 200 mg/kg de tioacetamida induziu a presença de células inflamatórias, todavia a lesão do parênquima hepático foi atenuada com o tempo. Portanto, a fim de se manter a constante lesão e inflamação do fígado, possivelmente será necessário aumentar a dose de TAA / Cirrhosis is a cosmopolitan disease that causes thousands of deaths each year and is in a state of attention by governmental health systems. In aid for its comprehension, we use the model of cirrhosis induced by thioacetamide (TAA) in rats. But this model has a lack of information about the presence of inflammatory cells in the liver, which play destructive action and stimulate the development of cirrhosis. We have proposed in this research to detect macrophages and dendritic cells in this model using 200mg/kg of thioacetamide via intraperitoneal for seven weeks and correlate them with apoptosis in hepatocytes, the presence of MCP-1 positive cells and the development of fibrosis in the liver of female rats. It was hoped to find steady increase in tissue and exudative macrophages and of dendritic cells attracted by the release of MCP-1 by hepatic stellate cells, to detect apoptosis in hepatocytes and consequent increase collagen deposition by hepatic stellate cells. The toxic action of thioacetamide was observed by piecemeal necrosis and tissue reaction with bile duct proliferation. The thioacetamide was effective in triggering the presence of macrophages and hepatocyte apoptosis in the first weeks, but ineffective in maintaining steadily increase of these events until the end of the seventh week. Furthermore, the increase of collagen tissue was not significant; indicating indirectly that the presence of macrophages and hepatocyte injury was not enough intense to trigger the maximum action of hepatic stellate cells during the seven weeks of treatment. The use of 200 mg / kg of thioacetamide induced the presence of inflammatory cells; however the injury of the hepatic parenchyma was attenuated with time.Therefore, in order to keep the constant liver injury and inflammation possibly will be necessary to increase the dose of TAA

Prostatacancer är en av världens vanligaste cancerformer. Varje år diagnostiseras ungefär 1,3 miljoner män världen över med prostatacancer och trots det vet man inte de bakomliggande orsakerna. Det finns flera studier som visar att det finns en koppling mellan infektion och olika typer av cancer och Cutibacterium acnes återfinns i hög utsträckning i prostatacancervävnad. Det har därför föreslagits att det finns ett samband mellan infektion av C. acnes och prostatacancer. Närvaro av tumörassocierade makrofager har visats sig vara gynnande för olika typer av cancer och från dessa makrofager frisätts kemokiner, bland annat CCL22. CCL22 kan vara inblandad i en lokal hämning av immunförsvaret som ofta förknippas med tillväxt av cancer. I detta arbete odlas makrofager från blodgivare med C. acnes för att ta reda på om makrofagerna ökar sitt uttryck av CCL22 mRNA. Genom att analysera resultaten från en kvantitativ realtids-PCR indikeras det att makrofager som behandlats med C. acnes signifikant ökar sitt uttryck av CCL22 mRNA. Sammanfattningsvis bedöms resultaten från detta arbete styrka uppfattningen att det kan finnas en koppling mellan C. acnes och orsakerna bakom prostatacancer men mer arbete återstår för att kunna klargöra dess relevans. / Prostate cancer is one of world´s most common forms of cancer. Every year about 1.3 million men around the world are diagnosed with prostate cancer and even so we cannot fully explain the etiology. There have been several studies indicating that there is a correlation between infection and different forms of cancer. Cutibacterium acnes (C.acnes) can be found to a large extent in prostate cancer tissue and a correlation between infection of C. acnes and prostate cancer has therefore been suggested. The presence of tumor-associated macrophages has been shown to favor various types of cancer. Several chemokines including CCL22 are released from these macrophages. CCL22 may be involved in a local inhibition of the immune system that is often associated with cancer growth. In this work, macrophages from a blood donor are grown with C. acnes to find out if the macrophages increase their expression of CCL22 mRNA after this exposure. By analyzing the results of a quantitative real-time PCR for CCL22 mRNA, it was demonstrated that macrophages treated with C. acnes significantly increase their expression of CCL22 mRNA. In conclusion, the results of this work indicate that there could be a link between C. acnes and the causes of prostate cancer, but more work remains to be able to clarify its relevance.

Considerando a importância do perfil do macrófago na infecção por Leishmania e o potencial papel de CD100 na modulação da ativação do m<font face=\"Symbol\">j, postulamos que essa molécula pode afetar a infectividade dessa célula por este parasita. Portanto, analisou-se e comparou-se a expressão de CD100 em diferentes tipos de m<font face=\"Symbol\">j (BALB/c e C57BL/6) e seus efeitos em termos de ativação e infecção com L. amazonensis. Avaliamos a expressão de CD100 e observamos que m<font face=\"Symbol\">js peritoneais de BALB/c expressam mais CD100 do que os de C57BL/6 e que esta expressão diminui ao longo da infecção. M<font face=\"Symbol\">j de BALB/c pré-tratados com sCD100 antes da infecção tem maior índice no tempo de 4h e quando em contato com o sCD100 antes, durante e após a infecção, o índice aumenta em 4 e 24h. Em m<font face=\"Symbol\">j de C57BL/6 pré-tratados com sCD100 ou no período de infecção, o índice aumenta em 4h. Quando permanecem durante todo o tempo em contato com o sCD100 o índice aumenta nos períodos de 4 e 24h. Concluímos que o CD100 pode estar relacionado com a modulação da infecção, e que a infecção afeta a expressão dessa molécula pelo m<font face=\"Symbol\">j. / Considering the importance of m<font face=\"Symbol\">j in Leishmania´s infection and the potential role of CD100 in the modulation of m<font face=\"Symbol\">j activation, we postulated that this molecule may affect m<font face=\"Symbol\">j infectivity. Therefore, we analyzed and compared the expression of CD100 on different types of m<font face=\"Symbol\">j (BALB/c and C57BL/6) and their effects in terms of m<font face=\"Symbol\">j activation and infectivity by L. amazonensis. We analyzed the expression of CD100 and the results suggest that m<font face=\"Symbol\">j from BALB/c express more CD100 than C57BL/6 and this expression decrease along the infection. M<font face=\"Symbol\">j from BALB/c pre-treated with sCD100 have augmented infection rates in 4h and in contact with the sCD100 before, during and after infection, increase rates in periods of 4 and 24h. In m<font face=\"Symbol\">j from C57BL/6 pre-treated with sCD100 or infection period, the rates increase in 4h. When macrophages remained in contact with sCD100 throughout the infection, rates increase after 4 and 24h. We can therefore conclude that CD100 may be related to modulation of Leishmania´s infection and that the infection itself affects CD100 expression in m<font face=\"Symbol\">j.