Thesis (Ph.D. in Immunology) -- University of Colorado at Denver and Health Sciences Center, 2006. / Typescript. Includes bibliographical references (leaves 182-208). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Orientador: James Venturini / Resumo: Paracoccidioidomicose (PCM) é uma micose sistêmica causada por fungos do gênero Paracoccidioides; suas principais formas clínicas são aguda/subaguda e crônica (FC). Restrita à América Latina, a PCM apresenta alta incidência no Brasil, Colômbia e Venezuela, especialmente entre os trabalhadores rurais. A maioria dos pacientes com a forma crônica da doença, mesmo após tratamento eficaz, apresentam sequelas, incluindo fibrose pulmonar (FP). Os problemas sociais, econômicos e psicológicos desencadeados pela FP são subestimados. Apesar da fibrogênese na PCM ser reconhecida como um processo precoce, seus mecanismos não estão totalmente conhecidos. Assim, o presente estudo tem por objetivo avaliar a influência de componentes dos isolados fúngicos e do hospedeiro sobre fibroblastos pulmonares. Foram utilizados fibroblastos pulmonares humanos da linhagem MRC-5 e fibroblastos pulmonares murinos, isolados de camundongos BALB/c. Fibroblastos foram cultivados in vitro com exoantígenos de isolados das espécies P. brasiliensis (Pb18 e Pb326) e P. lutzii (Pb01, Pb8334 e Pb66); gp43 purificada, e soros de pacientes com FC em diferentes momentos do tratamento (antes do tratamento, cura clínica, cura sorológica e cura aparente). Após 24 horas de cultivo, foram avaliadas a proliferação celular e produção de citocinas e fatores de crescimento por fibroblastos murinos e humanos. Nossos resultados demonstraram que componentes do fungo interferem em fibroblastos pulmonares, pela indução da proliferaç... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Made available in DSpace on 2014-06-11T19:33:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-11-21Bitstream added on 2014-06-13T20:05:11Z : No. of bitstreams: 1 fabro_at_dr_botfm.pdf: 9225853 bytes, checksum: cd0dd6d4aee86763e86640feaa8792e7 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A fibrose pulmonar é a base patológica de uma variedade de doenças crônicas incuráveis. A IL-17A, uma glicoproteína secretada de células Th17, é uma citocina pró-fibrótica relacionada recentemente à síntese de colágeno V e fibrose pulmonar. O remodelamento parenquimatoso pulmonar da matriz extracelular pelo colágeno I e V, apoptose e resposta Th17 foi estudado em camundongos Balb/c, C57/B6J selvagens e com knockout para o receptor A da IL-17; para determinar as vias fisiopatológicas que prolongam a fase tardia do processo fibrótico induzido por bleomicina e paraquat. Microscopia eletrônica, imunofluorescência, imunohistoquímica, detecção in situ da apoptose, morfometria, reconstrução tridimensional e reação em cadeia da polimerase em tempo real foram usados para avaliar a quantidade, estrutura e cadeias moleculares dos tipos de colágenos, apoptose e células imunes. Verificamos um aumento da síntese e secreção do colágeno V que promove a perpetuação da fibrose pulmonar de maneira IL-17A dependente. Além disso, observou-se que marcadores críticos da resposta Th17 como IL17, STAT3, TGF-β, IL-6, IL- 21, IL-23 e células T CD4+ foram significantemente aumentados em cepas susceptíveis a fibrose pulmonar e intensificadas na ausência do receptor A da IL-17. O aumento de marcadores Th17 resulta em um aumento de células T CD4+ através de uma resposta imune que efetivamente bloqueia a degradação do colágeno V, o qual contribui para o bloqueio da apoptose. Nosso estudo indica que o colágeno V participa da perpetuação da fibrose pulmonar por mecanismos IL-17 dependente e independentes, indicando o potencial alvo terapêutico do colágeno V e das vias de sinalização da resposta IL-17 no tratamento das doenças fibroproliferativas pulmonares / Pulmonary fibrosis is the pathologic basis for a variety of incurable human chronic lung diseases. IL-17A, a glycoprotein secreted from IL-17– producing cells, has recently been shown to be a profibrotic cytokine involved in type V collagen synthesis and pulmonary fibrosis. Remodeling of the extracellular matrix by collagen I and V, cell death and Th-17 immune response were evaluated in Balb/c, wild and IL-17 receptor A knockout C57/B6J mice, to determine the pathways that prolong the late phase of the fibrotic process induced by bleomycin and paraquat. Electron microscopy, immunofluorescence, immunohistochemistry, in situ detection of apoptosis, morphometry, tridimensional reconstruction and a real-time PCR were used to evaluate the amount, structure and molecular chains of collagen types, apoptosis and immune cells. We verified increased synthesis and secretion of type V collagen that promoted the maintenance of pulmonary fibrosis in a IL-17A dependent manner. However, we observed that the critical Th17 markers, IL-17, STAT3, TGF-β, IL-6, IL-21, IL-23 and CD4+ T cells, were significantly increased in the fibrosis-susceptible strain and intensified in the absence of IL-17 receptor A. Increased Th17 markers resulted in an increase in CD4+ T cells in fibrotic lung tissue toward an immune response that effectively blocked degradation of collagen V, which contributes to block apoptosis. Our studies indicate that collagen V participates in the maintenance of pulmonary fibrosis in both Th-17 – dependent and -independent manners and that collagen V and the components of the Th-17 signaling pathway are potential therapeutic targets for the treatment of fibroproliferative lung diseases

Made available in DSpace on 2014-06-11T19:23:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-28Bitstream added on 2014-06-13T19:29:05Z : No. of bitstreams: 1 felix_rg_me_botfm.pdf: 5628259 bytes, checksum: c9b3290ea27929658f6c6ebfde5ec2b6 (MD5) / A FPI é definida como um tipo de doença intersticial fibrosante crônica, de etiologia desconhecida, limitada aos pulmões e que apresenta padrão histológico de pneumonia intersticial usual. A prevalência da FPI é estimada em aproximadamente 20/100000 nos homens e 13/100000 nas mulheres, sendo que a média da idade quando do diagnóstico é de 67 anos e a sobrevida média é de 2 a 5 anos. Estima‐se cerca de 5 milhões de pessoas afetadas em todo o mundo. O tratamento clínico atual está associado à melhora parcial e transitória com resultados duvidosos ou insatisfatórios. A abordagem cirúrgica da FPI compreende o transplante de pulmão e é raro em decorrência da escassez de doadores e pela limitação de equipes capacitadas para realizar os procedimentos. A terapia celular apresenta‐se como uma alternativa terapêutica com grande potencial de aplicabilidade em fibrose pulmonar. Este estudo tem como objetivo a validação do modelo de fibrose pulmonar induzido pela instilação de bleomicina e a análise da contribuição da terapia celular utilizando a injeção de células‐tronco mesenquimais, alogênicas, obtidas de tecido adiposo para o tratamento de fibrose pulmonar. Foram utilizados 60 ratos machos, albinos, da raça Wistar, com peso médio de 250g com idade de 8 semanas. Foram constituídos 4 grupos experimentais compostos por 15 animais sendo monitorados: peso e o índice de saturação em 3 momentos: D0, D14 e D28. Em D0 foi realizada a instilação da bleomicina na dose de 1,5U/Kg. Em D14 a infusão das células tronco mesenquimais quantificadas e fenotipadas por citometria de fluxo. Em D28 foi realizado o sacrifício dos animais para análise macro e microscópica. A média de tecido adiposo retirado foi de 3,35g sendo que após a dissociação com o uso de colagenase tipo I a contagem celular foi de 5,3 x 104 células... / The IPF is defined as a type of chronic fibrosing interstitial lung disease of unknown etiology, limited to the lungs and that presents histological pattern of usual interstitial pneumonia. The prevalence of IPF is estimated at approximately 20/100000 in men and 13/100000 in women, and the mean age at diagnosis is 67 years and the median survival is 2 to 5 years. It is estimated about 5 million affected people worldwide. The current clinical treatment is associated with transient and partial improvement with equivocal or unsatisfactory. The surgical approach consists of IPF and lung transplantation is rare due to the scarcity of donors and by limiting teams trained to perform the procedures. Cell therapy is presented as an alternative therapy with a potential application in pulmonary fibrosis. This study aims to validate the model of pulmonary fibrosis induced by bleomycin instillation and analysis of the contribution of cell therapy using injection of mesenchymal stem cells, allogeneic, obtained from adipose tissue for the treatment of pulmonary fibrosis. A total of 60 male rats, albino Wistar rats, weighing 250g aged 8 weeks. 4 experimental groups were formed consisting of 15 animals being monitored: weight and saturation index in 3 stages: D0, D14 and D28. In D0 was performed instillation of bleomycin at a dose of 1.5 U / Kg. In D14 infusion of mesenchymal stem cells quantified and phenotyped by flow cytometry. In D28 was the sacrifice of animals for macro and microscopic analyzes. The mean adipose tissue removed was 3.35 g and that after dissociation using collagenase type I cell count was 5.3 x 104 lymphomononuclear cells/g of adipose tissue. These cells were expanded in culture for 21 days Knockout DMEM‐F12 medium added with 10% fetal bovine serum. Three criteria were considered proof of MSC: adhesion to plastic... (Complete abstract click electronic access below)

Orientador: Elenice Deffune / Banca: Lisete Ribeiro Teixeira / Banca: Hugo Hyung Bok Yoo / Resumo: A FPI é definida como um tipo de doença intersticial fibrosante crônica, de etiologia desconhecida, limitada aos pulmões e que apresenta padrão histológico de pneumonia intersticial usual. A prevalência da FPI é estimada em aproximadamente 20/100000 nos homens e 13/100000 nas mulheres, sendo que a média da idade quando do diagnóstico é de 67 anos e a sobrevida média é de 2 a 5 anos. Estima‐se cerca de 5 milhões de pessoas afetadas em todo o mundo. O tratamento clínico atual está associado à melhora parcial e transitória com resultados duvidosos ou insatisfatórios. A abordagem cirúrgica da FPI compreende o transplante de pulmão e é raro em decorrência da escassez de doadores e pela limitação de equipes capacitadas para realizar os procedimentos. A terapia celular apresenta‐se como uma alternativa terapêutica com grande potencial de aplicabilidade em fibrose pulmonar. Este estudo tem como objetivo a validação do modelo de fibrose pulmonar induzido pela instilação de bleomicina e a análise da contribuição da terapia celular utilizando a injeção de células‐tronco mesenquimais, alogênicas, obtidas de tecido adiposo para o tratamento de fibrose pulmonar. Foram utilizados 60 ratos machos, albinos, da raça Wistar, com peso médio de 250g com idade de 8 semanas. Foram constituídos 4 grupos experimentais compostos por 15 animais sendo monitorados: peso e o índice de saturação em 3 momentos: D0, D14 e D28. Em D0 foi realizada a instilação da bleomicina na dose de 1,5U/Kg. Em D14 a infusão das células tronco mesenquimais quantificadas e fenotipadas por citometria de fluxo. Em D28 foi realizado o sacrifício dos animais para análise macro e microscópica. A média de tecido adiposo retirado foi de 3,35g sendo que após a dissociação com o uso de colagenase tipo I a contagem celular foi de 5,3 x 104 células... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The IPF is defined as a type of chronic fibrosing interstitial lung disease of unknown etiology, limited to the lungs and that presents histological pattern of usual interstitial pneumonia. The prevalence of IPF is estimated at approximately 20/100000 in men and 13/100000 in women, and the mean age at diagnosis is 67 years and the median survival is 2 to 5 years. It is estimated about 5 million affected people worldwide. The current clinical treatment is associated with transient and partial improvement with equivocal or unsatisfactory. The surgical approach consists of IPF and lung transplantation is rare due to the scarcity of donors and by limiting teams trained to perform the procedures. Cell therapy is presented as an alternative therapy with a potential application in pulmonary fibrosis. This study aims to validate the model of pulmonary fibrosis induced by bleomycin instillation and analysis of the contribution of cell therapy using injection of mesenchymal stem cells, allogeneic, obtained from adipose tissue for the treatment of pulmonary fibrosis. A total of 60 male rats, albino Wistar rats, weighing 250g aged 8 weeks. 4 experimental groups were formed consisting of 15 animals being monitored: weight and saturation index in 3 stages: D0, D14 and D28. In D0 was performed instillation of bleomycin at a dose of 1.5 U / Kg. In D14 infusion of mesenchymal stem cells quantified and phenotyped by flow cytometry. In D28 was the sacrifice of animals for macro and microscopic analyzes. The mean adipose tissue removed was 3.35 g and that after dissociation using collagenase type I cell count was 5.3 x 104 lymphomononuclear cells/g of adipose tissue. These cells were expanded in culture for 21 days Knockout DMEM‐F12 medium added with 10% fetal bovine serum. Three criteria were considered proof of MSC: adhesion to plastic... (Complete abstract click electronic access below) / Mestre

Les cellules dendritiques, puissantes cellules présentatrices d'antigène, jouent un rôle-clé dans l'initiation et la régulation des réponses immunitaires et inflammatoires (Lipscomb et al., 2002). Elles sont présentes à l'état de cellules immatures dans la plupart des muqueuses, disséminées dans le tissu interstitiel et les épithéliums. Elles sont chargées de capter les antigènes exogènes, de les apprêter et de migrer vers les organes lymphoïdes pour les présenter aux lymphocytes T spécifiques. Au cours de cette migration, elles acquièrent les caractéristiques des cellules dendritiques matures présentes dans les organes lymphoïdes (expression forte des molécules HLA de classe II et des molécules de costimulation CD40, CD80, CD83, CD86). Les cellules dendritiques sont présentes dans le poumon normal (Vermaelen et al., 2005). Elles sont impliquées dans la physiopathologie de différentes maladies pulmonaires et jouent un rôle pathogénique essentiel dans certaines d'entre elles comme l'histiocytose langerhansienne ou l'asthme (Tazi et al ., 2000; Lambrecht et Hammad, 2003). Leur rôle au cours des processus fibrosants n'a cependant jamais été évalué. En effet, contrairement à d'autres cellules présentatrices d'antigènes que sont les macrophages, dont le rôle dans la réaction fibrosante est clairement établi (Agostini et al ., 1997), le rôle des cellules dendritiques au cours des processus de réparation alvéolaire n'a pas été étudié. Notre travail visait à étudier in vivo le rôle des cellules dendritiques au cours de la fibrose pulmonaire sur un modèle animal par induction d’une fibrose à la bléomycine chez la souris. Les premières étapes du projet ont consisté à caratériser par cytométrie en flux la présence de cellules dendritiques dans le poumon des animaux 3, 7 et 14 jours après instillation intratrachéale de bléomycine. L'étude s'est poursuivie par une caractérisation du phénotype de surface des cellules dendritiques en cherchant à préciser leur état d’activation et de maturation. L'étape suivante a consisté à rechercher par RT-QPCR les principales chimiokines responsables du recrutement des cellules dendritiques. Les résultats montrent une augmentation du nombre des cellules dendritiques CD11c+ / CMH II+ infiltrant le poumon dès J7, avec une sous-population de cellules activées exprimant fortement les molécules CMH II. Ces résultats sont corroborés par une augmentation de la population cellulaire totale du broyat de poumon dès J3, ainsi que par une augmentation de la cellularité totale et du nombre de cellules inflammatoires dans les lavages bronchoalvéolaires (LBA) à J7 et J14. L'étude a été complétée par une caractérisation plus approfondie du phénotype de surface des cellules dendritiques en cherchant à préciser leur état d’activation/maturation. [...] / Dendritic cells (DCs), potent antigen-presenting cells, play a key role in the initiation and regulation of immune and inflammatory responses (Lipscomb et al., 2002). Immature DCs are present in the most of mucous membranes, disseminated in the interstitial tissue and the epithelia. They are able to deal with exogenous antigens, to process them and to migrate to lymphoid organs and to present them to T lymphocytes. During their migration, they have the characteristics of mature DCs in lymphoid organs (higher expression of MHC class-II molecules and costimulation-molecules (CD40, CD80, CD83, CD86). Dendritic cells are present in the normal lung (Vermarlen et al., 2005). They are implicated in the pathophysiology of different pulmonary diseases and play a crucial pathogenic role in some of them like Langerhan’s cell Histiocytosis or asthma (Tazi et al., 2000; Lambrecht et Hammad, 2003). However, their role in the fibrosing process has never been studied. In fact, the role of macrophages (other antigen presenting cells (APCs)) in fibrosing reaction has been clearly established (Agostini et al., 1997), but DCs function during the alveolar healing process has not been studied. The purpose of this thesis is to study the in vivo role of DCs in bleomycin-induced pulmonary fibrosis in mice. The first step of this project is to demonstrate by flow cytometry the presence of DCs in the lung of these animals 3, 7 and 14 days after intratracheal bleomycin instillation. We continued our research with the surface phenotypic characterization of DCs identifying their activation state and maturation. The next step consists to search the main chemokines which are responsive for dendritic cells recruitment by RT-qPCR. Our results show an increase of CD11c+ / MHC class II+ DCs number infiltrating the lung after D7, with an activated cell subpopulation which strongly express MHC class II molecules. The results are corroborated by an increase of the total cell population in the lung homogenate after D3 and by an increase of the total cellularity and inflammatory cells number in broncho-alveolar lavages (BAL) at D7 and at D14. This study was completed by the surface phenotypic characterization of DCs identifying their activation state and maturation. [...]

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第19573号 / 医博第4080号 / 新制||医||1013(附属図書館) / 32609 / 京都大学大学院医学研究科医学専攻 / (主査)教授 伊達 洋至, 教授 小池 薫, 教授 瀬原 淳子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 新制・課程博士 / 博士(社会健康医学) / 甲第24808号 / 社医博第132号 / 新制||社医||12(附属図書館) / 京都大学大学院医学研究科社会健康医学系専攻 / (主査)教授 川上 浩司, 教授 西浦 博, 教授 平井 豊博 / 学位規則第4条第1項該当 / Doctor of Public Health / Kyoto University / DFAM

No description available.

Doctor of Philosophy / Department of Human Nutrition / Richard C. Baybutt / Weiqun George Wang / Although omega-3 fatty acids have been a hot issue in nutrition for years, there remains a paucity of research on the topic of omega-3 fatty acid and pulmonary fibrosis and the mechanism is still unclear. The purpose of this research is to investigate the preventive effects of flaxseed oil for bleomycin-induced pulmonary fibrosis in rats and to find the possible underlying mechanisms. There are two experiments demonstrated in this dissertation, one is with various doses of flaxseed oil in the diet (0, 2.5, 5, 7.5, 10, 12.5, and 15 % (w/w)), and the other is with different times of sacrificing animals after oropharyngeal bleomycin treatment (days 7 and 21). In the first study, three proteins including transforming growth factor-[beta] (TGF-[beta]), interleukin-1 (IL-1), and [alpha]-smooth muscle actin ([alpha]-SMA), commonly associated with fibrotic inflammation in the lung, were examined by Western blot and fatty acids composition of the diets and tissues were analyzed by gas chromatography (GC). Fifteen percent of flaxseed oil group significantly reduced septal and vascular thickness and fibrosis in the lung, and significant cardiac fibrosis in the heart. The amount of IL-1 and [alpha]-SMA decreased significantly as the amount of omega-3 fatty acids increased, whereas TGF-[beta] did not change significantly. The next study further reported the time-course effect and potential underlying mechanisms. Both interleukin-6 (IL-6), a protein associated with fibrotic inflammation in the lung, and renin, an enzyme related to renin-angiotensin system, were examined by Western blot. The time-dependent increase of IL-6 in response to bleomycin treatment was reversed by flaxseed oil diet. Although renin was not significantly different in the kidney, it suggested that the renin-angiotensin system may be involved locally. In addition, the profiles of fatty acids in both liver and kidney tissues as measured by lipidomics demonstrated a significant increase of omega-3: omega-6 ratio in the flaxseed oil-fed groups. Overall, these results indicated for the first time that the omega-3 fatty acids rich in flaxseed oil inhibited the formation of pulmonary fibrosis in a dose-dependent manner - however the moderate dose of flaxseed oil was most effective - via anti-inflammatory mechanisms, which appears associated with the modulated fatty acid composition in the tissues.