La biosynthèse de phénol-glycolipides (PGL) par Mycobacterium tuberculosis et M. leprae favorise l’invasion des macrophages via l'interaction de la partie saccharidique des PGL avec le domaine lectine du récepteur cellulaire au complément CR3. Les PGL inhibent également la production de cytokines inflammatoires par la cellule hôte, par un mécanisme inconnu. J’ai observé que des bactéries BCG transgéniques exprimant les PGL de M. tuberculosis ou M. leprae avaient une capacité de survie accrue dans les macrophages. Cette persistance intracellulaire était dépendante de CR3 et associée à une diminution de la production d'oxyde nitrique dans les cellules infectées. L’addition de PGL purifié suffisait à inhiber la production d’oxyde nitrique par des macrophages stimulés avec LPS/IFN-γ. J’ai montré que la liaison de PGL-1 à CR3 provoquait la dégradation post-transcriptionnelle de TIR-domain-containing adapter-inducing interféron-β (TRIF) dans les macrophages, ce qui entraînait une réduction de la signalisation TRIF-dépendante. Dans les macrophages stimulés avec LPS/IFN-γ, la dégradation de TRIF réduisait la production d’oxyde nitrique synthase, et la production TRIF dépendante de cytokines inflammatoires et des chimiokines. Mes résultats ont donc permis d’identifier un nouveau mécanisme de virulence développé par les mycobactéries pathogènes pour réprimer conjointement les réponses inflammatoires et antimicrobiennes de l’hôte / Biosynthesis of phenolic glycolipids (PGL) by Mycobacterium tuberculosis and M. leprae promotes macrophage invasion, which proceeds through the interaction of the PGL sugar moieties with the lectin domain of cell-displayed complement receptor (CR3). PGL also limit host cell production of inflammatory cytokines by an unknown mechanism. I observed that transgenic BCG that express PGL specific to M. tuberculosis or M. leprae displayed enhanced survival within macrophages. Increased intracellular persistence of PGL-expressing BCG was CR3-dependent and correlated with the decreased production of nitric oxide in infected cells. Notably, the addition of soluble PGL to macrophages was sufficient to induce a reduction in nitric oxide production upon stimulation with LPS/IFN-γ. I showed that PGL-1 binding to CR3 causes the post-transcriptional degradation of TIR-domain-containing adapter-inducing interferon-β (TRIF) in macrophages, resulting in impaired TRIF-dependent signaling. Functionally, PGL-1-mediated degradation of TRIF resulted in the decreased induction of nitric oxide synthase, and TRIF-dependent inflammatory cytokines and chemokines in LPS/IFN-γ-stimulated macrophages. My results thus identified a virulence mechanism evolved by pathogenic mycobacteria to suppress both the inflammatory and antimicrobial responses of infected host cells

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Endotoxins (E) are a unique and abundant family of glycolipids located in the outer leaflet of the outer membrane of Gram-negative bacteria. Host immune responses to endotoxin depend on ordered endotoxin-host protein interactions, resulting in delivery of an endotoxin monomer to MD-2 which acts as a potent agonist of Toll-Like Receptor (TLR) 4. Activated TLR4 is unique among TLRs in its ability to mobilize two distinct intracellular signaling pathways: the MyD88- and TRIF-dependent pathways. The regulated action of both pathways is likely important for optimal host immune responses to Gram-negative bacterial infection, but how this is achieved is not well understood Recent studies have indicated an essential role for host CD14 in TRIF-dependent signaling by activated TLR4 but the extent to which these observations reflect a general role of CD14 in endotoxin-triggered TRIF signaling or one more narrowly restricted to the specific endotoxins and/or cell types used is uncertain. We have addressed this question by identifying a novel CD14-independent mechanism for efficient delivery of E monomer to MD-2 and TLR4 activation, that is mediated by endotoxin.albumin complexes. We have used these complexes to demonstrate CD14-independent activation of MD-2⋅TLR4 by a wider range of endotoxin species than previously thought possible and activation of both MyD88- and TRIF-dependent pathways. Taken together, the findings in this thesis indicate that the molecular structure and physical presentation of endotoxin as well as CD14-independent properties of the host cell help determine the extent to which CD14 is required for TRIF-dependent signaling by activated TLR4.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

京都大学 / 0048 / 新制・課程博士 / 博士(生命科学) / 甲第22595号 / 生博第428号 / 新制||生||57(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 藤田 尚志, 教授 松田 道行, 教授 杉田 昌彦 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

Certains motifs moléculaires associés à la mort cellulaire semblent identifier les cancers prompts à répondre à une thérapie cytotoxique. Ceci en élaborant une réponse anti-tumorale basées sur une réponse T protectrice. Mon travail de thèse montre que le traitement par chimiothérapie immunogène active des voies moléculaires mimant une infection virale. Ceci conduit au niveau des cellules tumorales à une signalisation autocrine via l’IFNαβ / IFNAR1/2, initiée par la reconnaissance d’ARN double brin (dsRNA) endogène par les Récepteurs endosomaux de Reconnaissance des Motifs (PRRs). De façon plus détaillée, nous montrons que les axes TLR3/TRIF (senseurs endosomaux de dsRNA) et IFNAR1/2 (Récepteurs de l’IFN de Type I) doivent signaliser au niveau de la cellule tumorale pour que la chimiothérapie puisse aboutir à l’induction de l’axe CXCL10/CXCR3 et éliciter une réponse efficace in vivo. L’analyse du profil ARN de cellules tumorales Tlr3+/+ (mais pas Tlr3-/-) exposées aux anthracyclines a révélé une forte empreinte virale/IFN, indispensable à l’efficacité/activité anti-tumorale. Le fait d’affecter les axes TLR3 ou IFNAR1/2 au niveau tumorale soit à l’aide d’anticorps neutralisants, soit à l’aide de modèles KO abroge le relarguage de CXCL10 induit par la chimiothérapie, et ainsi la capacité à contrôler la pousse tumorale à moins que de l’IFNαβ ou du CXCL10 exogène soit co-administré aux anthracyclines. De plus la chimiorésistance des tumeurs traitées par des molécules n’induisant pas de signature virale peux être réversée par de l’IFN de Type I exogène. Enfin, la détection d’une signature IFN au niveau de biospies de cancers du sein humains permet de prédire la bonne réponse au traitement adjuvant par anthracyclines. D’un point de vue de l’évolution, alors que les tumeurs (comme les virus) ont élaboré des mécanismes pour échapper aux réponses IFN, la signature virale induite par la chimiothérapie devrait contribuer à contrecarrer cette immunoédition. / Distinct cell death-associated molecular patterns might define cancers proned to respond to a cytotoxic therapy by mounting a protective T cell-based anticancer immunity. My PhD Thesis work shows that immunogenic chemotherapy phenocopies viral infection leading to autocrine IFNαβ/IFNAR1/2 signalling in tumor cells initiated by recognition of self dsRNA by endosomal pattern recognition receptors (PRRs). In detail, TLR3/TRIF (endosomal dsRNA sensors) and IFNAR1/2 (Type I IFN receptors) must signal within the tumor cells so that chemotherapy can induce downstream CXCL10/CXCR3 axis and elicit therapeutic responsiveness in vivo. RNA profiling of Tlr3+/+ (but not Tlr3-/-) tumor cells exposed to anthracyclines revealed a strong IFN/viral fingerprint, indispensable for the tumoricidal activity. Neutralization by antibodies or genetic defects affecting tumor –associated TLR3 or IFNAR1/2 compromised chemotherapy-induced CXCL10 release and tumor control unless exogenous IFNαβ or CXCL10 are concomitantly supplied to anthracyclines. Moreover, chemoresistance of tumors treated by drugs failing to induce a viral signature can be reversed by exogenous Type I IFN. Finally, the IFN fingerprint of human breast cancers allowed to predict tumors proned to benefit from adjuvant anthracyclines. From an evolutionary viewpoint, while tumors (like viruses) have evolved mechanisms to evade an IFN response, chemotherapy-induced viral mimicry might contribute to bypass such as immunoediting.

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Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Made available in DSpace on 2014-12-17T14:55:19Z (GMT). No. of bitstreams: 1 PauloCSC_capa_ate_pag_14.pdf: 6010526 bytes, checksum: e2d3959764645f15a68a651383c52a92 (MD5) Previous issue date: 2002-05-10 / This work has as main objective to show all the particularities regarding the Three-phase Power Summation Method, used for load flow calculation, in what it says respect to the influence of the magnetic coupling among the phases, as well as to the losses presented in all the existent transformers in the feeder to be analyzed. Besides, its application is detailed in the study of the short-circuits, that happen in the presence of high impedance values, which possess a problem, that is its difficult detection and consequent elimination on the part of common devices of protection. That happens due to the characteristic presented by the current of short? circuit, in being generally of the same order of greatness that the load currents. Results of simulations accomplished in several situations will be shown, objectifying a complete analysis of the behavior of the proposed method in several types of short-circuits. Confront of the results obtained by the method with results of another works will be presented to verify its effectiveness / Este trabalho tem como objetivo principal mostrar todas as particularidades a respeito do M?todo da Soma de Pot?ncias Trif?sico, utilizado em c?lculo de fluxo de carga trif?sico, no que diz respeito ? influ?ncia do acoplamento magn?tico entre as fases, bem como ?s perdas apresentadas em todos os transformadores existentes no alimentador a ser analisado. Al?m disso, detalha-se a sua aplica??o no estudo dos curtos-circuitos, que ocorrem na presen?a de altos valores de imped?ncia, os quais possuem um problema a mais, que ? sua dif?cil detec??o e conseq?ente elimina??o por parte de dispositivos comuns de prote??o. Isso ocorre devido ? caracter?stica apresentada pela corrente de curto, em ser geralmente da mesma ordem de grandeza que as correntes de carga. Ser?o mostrados resultados de simula??es realizadas em diversas situa??es, buscando uma an?lise completa do comportamento do m?todo proposto em v?rios tipos de curtos-circuitos. Confronto dos resultados obtidos pelo m?todo com os de outros trabalhos ser?o apresentados como forma de verifica??o de sua efic?cia

Made available in DSpace on 2014-12-17T14:55:42Z (GMT). No. of bitstreams: 1 LucianoPSJ_DISSERT.pdf: 6664942 bytes, checksum: fc1840aaaecdac8b8558e271f979adbf (MD5) Previous issue date: 2009-04-17 / This work deals with the development of an experimental study on a power supply of high frequency that provides the toch plasmica to be implemented in PLASPETRO project, which consists of two static converters developed by using Insulated Gate Bipolar Transistor (IGBT). The drivers used to control these keys are triggered by Digital Signal Processor (DSP) through optical fibers to reduce problems with electromagnetic interference (EMI). The first stage consists of a pre-regulator in the form of an AC to DC converter with three-phase boost power factor correction which is the main theme of this work, while the second is the source of high frequency itself. A series-resonant inverter consists of four (4) cell inverters operating in a frequency around 115 kHz each one in soft switching mode, alternating itself to supply the load (plasma torch) an alternating current with a frequency of 450 kHz. The first stage has the function of providing the series-resonant inverter a DC voltage, with the value controlled from the power supply provided by the electrical system of the utility, and correct the power factor of the system as a whole. This level of DC bus voltage at the output of the first stage will be used to control the power transferred by the inverter to the load, and it may vary from 550 VDC to a maximum of 800 VDC. To control the voltage level of DC bus driver used a proportional integral (PI) controller and to achieve the unity power factor it was used two other proportional integral currents controllers. Computational simulations were performed to assist in sizing and forecasting performance. All the control and communications needed to stage supervisory were implemented on a DSP / O presente trabalho trata do desenvolvimento de um estudo experimental sobre uma fonte de alimenta??o de alta freq??ncia que alimentar? a tocha pl?smica a ser implementada no projeto PLASPETRO, o qual consta de dois conversores est?ticos desenvolvidos com o uso de transistores - Insulated Gate Bipolar Transistor (IGBT). Os drivers utilizados no controle dessas chaves s?o acionados por um controlador de sinal digital - Digital Signal Processor (DSP) atrav?s de fibras ?pticas a fim de reduzir problemas com interfer?ncia eletromagn?tica - Electromagnetic interference (EMI). O primeiro consiste de um est?gio pr?-regulador na forma de um conversor CA-CC boost trif?sico com corre??o de fator de pot?ncia, tema principal deste trabalho, enquanto o segundo consiste na fonte de alta frequ?ncia propriamente dita. Um inversor s?rie-ressonante composto por 4 (quatro) c?lulas inversoras operando a uma frequ?ncia de cerca de 115 kHz cada, em modo de comuta??o suave, alternando-se para fornecer ? carga (tocha de plasma) uma corrente alternada com uma frequ?ncia de 450 kHz. O primeiro est?gio tem a fun??o de fornecer ao inversor s?rie-ressonante uma tens?o CC de valor controlado a partir da rede de alimenta??o disponibilizada pelo sistema el?trico da concession?ria, al?m de corrigir o fator de pot?ncia do sistema como um todo. Esse n?vel CC de tens?o no barramento de sa?da do primeiro est?gio ser? utilizado para o controle da pot?ncia transferida pelo inversor ? carga, podendo variar desde 550Vcc at? um valor m?ximo de 800Vcc. Para controlar o n?vel de tens?o do barramento CC utilizou-se um controlador Proporcional Integral (PI) e para atingir o fator de pot?ncia unit?rio utilizou-se dois outros PIs para controle das correntes em referencial estacion?rio. Simula??es computacionais foram realizadas para auxiliar no dimensionamento e previs?o de performance. Todo o controle e comunica??es necess?rias ao est?gio supervis?rio foram implementadas em um DSP