Das Ergebnis einer T-Zellmediierten Immunantwort ist von der Signalvermittlung durch kostimulatorische Moleküle abhängig. Diese kostimulatorischen Moleküle sind - neben dem spezifischen Antigen - notwendig für eine vollständige T-Zellaktivierung, die es der T-Zelle erlaubt zu proliferieren, neue Oberflächenantigene zu exprimieren und Zytokine zu sezernieren. Ohne das kostimulatorische Signal wird die T-Zelle anerg oder sogar apoptotisch, eine effektive Immunantwort ist dann nicht möglich. Die vorliegende Dissertationsschrift enthält die initiale Beschreibung eines neuen kostimulatorischen Moleküls, eine umfangreiche Charakterisierung seiner Expression in vitro, sowie seiner Funktion. Das Molekül "ICOS" (ICOS steht für inducible costimulator) ist ein T-Zellspezifisches Molekül und weist eine große Homologie zu dem Prototyp eines kostimulatorischen Moleküls, dem CD28 Molekül auf. Die Experimente, die zur Charakterisierung der Induktionsbedingungen des ICOS Moleküls durchgeführt wurden, zeigen, daß die Expression des ICOS Moleküls sehr schnell nach T-Zellaktivierung induziert wird, die Expressionsstärke innerhalb von Stunden stark heraufreguliert wird und die Expression lange (mindestens 96h) auf der T-Zelloberfläche zu detektieren ist. Ein Vergleich mit anderen aktivierungsabhängigen T-Zelloberflächenmolekülen zeigt eine ICOS-spezifische Zeitkinetik der Expression, die durch verschiedene T-Zellstimuli zu induzieren ist. Eine optimale Expression des Moleküls ist zwei-signalabhängig und durch Cyclosporin A blockierbar. Bezüglich der Funktion von ICOS wurde die Wirkung der Kostimulation via ICOS auf eine Reihe von kritischen Parametern der T-Zellaktivierung analysiert. Durch die Kostimulation mit einem ICOS-spezifischen Antikörper wird konzentrationsabhängig die T-Zellproliferation induziert, unabhängig davon, ob das ersten Signal via CD3 oder via einen löslichen Stimulus, wie PHA erfolgte. Die ICOS Kostimulation bewirkt die Hochregulation von typischen T-Zellaktivierungsantigenen und sie induziert oder verstärkt die Sekretion zahlreicher Lymphokine. Die verstärkende Wirkung auf alle diese Parameter der T-Zellaktivierung führt schließlich dazu, daß die über ICOS kostimulierten Zellen in der Lage sind T-Zellhilfe für B-Zellen zu leisten, so daß diese Immunglobuline sezernieren. Wurde versucht, die direkte Interaktion des ICOS Moleküls mit seinem potentiellen Liganden bei der T-Zellinduzierten Immunglobulinsekretion durch den mAk F44 zu blockieren, so zeigte sich kein Effekt. Bei Langzeitkostimulation via ICOS zeigte sich allerdings, daß die Kostimulation durch das ICOS Molekül auch in der Lage ist, die T-Zellaktivierung negativ zu beeinflussen: Die Langzeitstimulation via ICOS erzeugt eine deutliche Proliferationsdepression und einen Viabilitätsverlust der T-Zellen. Insgesamt lassen diese Ergebnisse den vorsichtigen Schluß zu, daß das ICOS Molekül eine wichtige Rolle an der Schnittstelle zwischen Expansion und Effektorfunktion einerseits und Depression der T-Zellen andererseits spielt. / The outcome of T-cell resonses after T-cell encounter with specific antigens is modulated by co-stimulatory signals, which are required for both lymphocyte activation and development of adaptive immunity. Here I report the initial characterization of a novel co-stimulatory molecule which enhances all basic T-cell functions and displays a unique induction and expression pattern. ICOS (for inducible co-stimulator) is a T-cellspecific activation antigen with high homology to CD28, the prototype of a co-stimulatory molecule. Analysis of induction requirements for ICOS expression revealed a two-signal dependency and cyclosporine A sensitivity. ICOS' expression kinetics are unique when compared with other early T-cell activation antigens. ICOS is induced very quickly on the T-cell surface and is rapidly upregulated following T-cell activation. The surface expression of ICOS is surprisingly prolonged - lasting at least 96 hours - considering its rapid induction kinetics. Stimulation via an ICOS-specific monoclonal antibody enhances all basic T-cell functions such as proliferation, upregulation of molecules that medicate cell-cell interaction, secretion of lymphokines and effective help for antibody secreting B-cells. Costimulation via ICOS is effective regardless of the route of action of the first signal (immobilized or soluble). Blockade of the ICOS interaction with its presumed ligand on B-cells does not inhibit immunoglobulin production by these B-cells, though. Finally long-term stimulation experiments reveal a possible negative role for ICOS in regulating T-cell responses. These results indicate that ICOS is a major regulator of the adaptive immune system determining the healthy balance of negative and positive signaling during T-cell activation and differentiation.

Der Induzierbare Kostimulator (ICOS) ist ein wichtiger Regulator der T-Zell-Effektorfunktion. In vivo führt ein Defekt von ICOS zur Beeinträchtigung der T-Zellabhängigen humoralen Immunität. In gendefizienten Mäusen wurden stark gestörte B-Zellantworten beobachtet. Mehrere in vitro und in vivo Studien führen diese Phänomene auf eine beeinträchtigte Regulation von Kommunikationsmolekülen auf der Zelloberfläche und Expression von Zytokinen durch ICOS-defiziente T-Zellen zurück. In dieser Arbeit konnte jedoch anhand Antigen-spezifischer T-Zellen in einem murinen adoptiven Transfersystem gezeigt werden, dass das Signal über ICOS die frühe T-Zellaktivierung nicht signifikant beeinflusst. Stattdessen trägt ICOS wesentlich zum Überleben und zur Expansion von Effektor T-Zellen bei, die zuvor lokal durch Antigengabe mit Adjuvanz induziert wurden. Diese beobachtete biologische Funktion von ICOS lässt sich auch auf FoxP3+ Regulatorische T-Zellen übertragen, welche durch systemische Antigengabe ohne Adjuvanz generiert wurden. In Übereinstimmung mit diesem Befund führt die Abwesenheit von ICOS unter homöostatischen Bedingungen in nicht-immunisierten Mäusen zu reduzierten Zellzahlen von Effektor-Memory T-Zellen und FoxP3+ Regulatorischen T-Zellen. Der regulierende Effekt von ICOS auf die Größe einer spezifischen Effektor T-Zellpopulation gilt auch für Follikuläre T-Helferzellen, konnte jedoch für zytotoxische CD8+ T-Zellen nicht eindeutig nachgewiesen werden. Auf der Grundlage dieser Ergebnisse kristallisiert sich eine globale biologische Rolle von ICOS für Effektorzellen heraus. Als kostimulatorisches, agonistisches Molekül reguliert ICOS generell die Pool-Größe aller Effektor T-Zellen mit unterschiedlichen, teilweise gegensätzlichen funktionellen Eigenschaften. Mit Hilfe dieses neuen Konzeptes können frühere in vivo Studien, deren Ergebnisse in Bezug auf die Funktion von ICOS scheinbar widersprüchlich waren, in Einklang gebracht werden. / The Inducible Co-Stimulator (ICOS) is an important regulator of T cell effector function. In vivo ICOS deficiency results in impaired T-cell dependent humoral immunity. Knock out mice show strongly defective B cell responses. Several in vitro and in vivo studies attributed this phenomenon to impaired upregulation of cell surface communication molecules and cytokine synthesis by ICOS-deficient T cells. However, in this work now could be shown with antigen-specific T cells in a murine adoptive transfer system that signaling via ICOS does not significantly affect early T cell activation. Instead, ICOS substantially contributes to the survival and expansion of effector T cells upon local challenge with antigen and adjuvant. Importantly, the observed biological function of ICOS also extends to FoxP3+ regulatory T cells, as can be observed after systemic antigen delivery without adjuvant. In line with these findings, absence of ICOS under homeostatic conditions of nonimmunized mice leads to a reduced number of both effector-memory and FoxP3+ regulatory T cells. The regulatory function of ICOS to control the poolsize of special T cell effector populations is also observed for follicular B helper T cells. The influence of ICOS on cytotoxic CD8+ T cells could not be clearly demonstrated. Based on these results, I propose a biological role for ICOS as a costimulatory, agonistic molecule for a variety of effector T cells with differing and partly opposing funtional roles. This concept may reconcile a number of past in vivo studies with seemingly cotradictory results on ICOS function.

Der induzierbare Kostimulator ICOS ist ein zu CD28 strukturell und funktionell verwandtes Molekül, das eine wichtige regulatorische Rolle bei der T-Zelleffektorfunktion spielt. Eine ICOS-Defizienz beim Mensch manifestiert sich in einer schweren Störung des humoralen Immunsystems. Eine murine ICOS-Defizienz führt ebenfalls zu einer Beeinträchtigung der T-Zell-abhängigen humoralen Immunantwort, bei der kleinere oder komplett fehlende Keimzentren zu beobachten sind. Vielfältige in vitro und in vivo Studien führten diese Phänomene auf die beeinträchtigte Regulation von Kommunikationsmolekülen der Zelloberfläche und der Zytokinexpression durch ICOS-defiziente T-Zellen zurück. Ein Ziel dieser Arbeit war es, mit Hilfe von ICOS KO Mäusen den Einfluss von ICOS auf die B-Zellentwicklung genauer zu untersuchen. Dabei konnte gezeigt werden, dass ICOS erst in der späten Phase der B-Zellentwicklung eine Rolle spielt, da der Interaktionspartner von ICOS erst auf transitionellen B-Zellen der Milz exprimiert wird. Durch die Etablierung eines in vivo adoptiven T-B Transfermodells konnte die Rolle von ICOS erstmalig bei der T-B Kooperation in den frühen Phasen der Immunantwort auf der Ebene Antigen-spezifischer T- und B-Zellen aufgeklärt werden. Dabei konnte beobachtet werden, dass eine ICOS-Defizienz einen dramatischen Einfluss auf die B-Zellexpansion und B-Zellproliferation hat. Zum ersten Mal konnte in vivo gezeigt werden, dass ICOS bei der T-B Kooperation eine entscheidende Rolle bei der Regulation diverser Oberflächenmarker der B-Zellen spielt, wodurch die B-Zellaktivierung, B-Zellproliferation und B-Zelldifferenzierung bei der Keimzentrums- und Plasmazellreaktion beeinflusst werden. Histologische Analysen zeigten, dass bei einer ICOS-Defizienz follikuläre T-Helferzellen nicht in die Keimzentrumsumgebung einwandern und daher keine T-Zellhilfe für die B-Zellen anbieten können. Dadurch kann die Keimzentrumsreaktion nicht weiter aufrechterhalten werden und eine Ausbildung von kleineren Keimzentren ist die Folge. Weiterhin konnte beobachtet werden, dass eine fehlende ICOS-Interaktion zwischen T- und B-Zellen zu einer Störung der Plasmazellgenerierung führt, wodurch auch die Mengen an messbaren Serumimmunglobulinen beeinflusst werden. Eine erhöhte Gabe von ICOS-defizienten T-Zellen kann diese Effekte nicht vollständig ausgleichen. Daher ist erkennbar, dass ICOS eine Vielzahl von zusätzlichen Faktoren beeinflusst, die für die ICOS-abhängigen B-Zelleffekte verantwortlich sind. / The inducible costimulator ICOS, structural and functional similar to CD28, plays an important regulatory role in T cell receptor function. The ICOS deficiency in humans is described as a severe dysfunction of the humoral immune response, resulting in dramatic reduced B cell numbers and impaired antibody response against pathogens. The murine ICOS-deficiency also leads to a disturbed T cell dependent immune response resulting in a reduced germinal center formation. Various in vitro and in vivo studies attributes this phenomenon to impaired upregulation of cell surface communication molecules and cytokine synthesis by ICOS-deficient T cells. In this work the investigations with ICOS KO mice should clarify the impact of ICOS in B cell development. As observed, ICOS can only play a role in the late phase B cell development, because the interaction partner is expressed on transitional B cells in the spleen. The establishment of an in vivo adoptive T-B transfer system could determine for the first time the role of ICOS in T-B cooperation in early immune response stages on antigen specific T and B cell levels. As shown, ICOS deficiency influences in a dramatic extend the B cell expansion and B cell proliferation. For the first time in vivo, we could demonstrate that ICOS plays a significant role by influencing the regulation of various B cell surface markers, which affects the B cell activation, B cell proliferation and B differentiation in germinal center or plasma cell reaction. Histological investigations revealed in the ICOS-deficiency that follicular T helper cells could not migrate into the germinal center microenvironment and therefore could not provide T cell help for B cells. As a result, the germinal center reaction could not maintained and therefore the formation of little germinal centers occurred. The missing interaction between T and B cells leads to a dysfunction in plasma cell generation and also influences the detectable amounts of serum immunglobulines. An administration of higher ICOS KO T cell numbers could not fully compensate these effects. Therefore, ICOS bias multitudes of additional factors, which are responsible for the ICOS dependent B cell effects.

Depuis la révolution industrielle, les croissances économique et démographique ont augmenté de manière exponentielle induisant l’augmentation de la combustion d’énergies fossiles, telles que le charbon, le pétrole, et le gaz naturel. La combustion de ces sources d’énergie conduit à l’émission de gaz à effet de serre, principalement le dioxyde de carbone (CO2) et le méthane (CH4), qui par leur accumulation dans l’atmosphère entraînent une augmentation de l’effet de serre. Selon le GIEC (Groupe d'experts Intergouvernemental sur l'Évolution du Climat), l’implication des émissions anthropiques dans l’augmentation de l’effet de serre est extrêmement probable avec un pourcentage de certitude qui dépasse 95%. Toutefois, l’estimation des bilans régionaux d'émissions de GES reste très incertaine. L’objectif de cette thèse est de contribuer à l’amélioration de l’estimation des bilans régionaux de GES en France, en utilisant pour la première fois les concentrations atmosphériques du CO2 et de CH4 mesurées par le réseau ICOS (Integrated Carbon Observation System) et la modélisation inverse à l’échelle régionale.Dans un premier temps, on s’est focalisé sur l’étude des concentrations mesurées de CO2, CH4 et CO (monoxyde de carbone) fournis par des stations de surface. Cette étude a pour objectif l'identification des mesures atmosphériques contaminées par les émissions locales (quelques kilomètres au tour de la station) et qui provoque ce qu’on appelle « les pics de concentrations ». Trois méthodes ont été appliquées sur des séries temporelles fournies par quatre stations du réseau ICOS, afin de déterminer leur degré de contamination. Les résultats des différentes méthodes ont été comparés entre eux, puis comparés à un inventaire de données contaminées préparé manuellement par les gestionnaires des stations. Cette comparaison a permis l’évaluation de la performance des trois méthodes pour la détection réussie des pics. À l’issue de ce travail, la méthode la plus performante a été proposée pour effectuer un nettoyage automatique des séries de mesure du réseau ICOS.Dans un deuxième temps, le modèle régional de chimie-transport CHIMERE est utilisé pour simuler les concentrations atmosphériques du CO2 et du CH4 de l’année 2014 sur un domaine centré sur la France. L’objectif de cette étude est d’étudier la sensibilité des concentrations simulées en utilisant différentes données d’entrées. Premièrement, on étudie la sensibilité des concentrations simulées par rapport au transport en utilisant deux modèles météorologiques AROME et ECMWF. Deuxièmes, on analyse la sensibilité des concentrations simulées face aux différentes cartes d’émissions. Dans cette dernière étape, on étudie les différences entre les cartes d’émissions anthropiques séparément des cartes d’émissions biogéniques. Ce travail nous permet de quantifier à la fois les erreurs liées aux transports et les erreurs liées aux flux d’émissions. La meilleure combinaison des données d’entrée va être sélectionnée pour l’étape d’inversion des flux.Dans un dernier plan, les mesures atmosphériques des concentrations de CO2 et du CH4 sont utilisées par le système d’inversion PYMAI (Berchet et coll., 2013 et 2015) afin d’estimer les bilans régionaux d'émissions des principaux GES en France. L’inversion s’est exécutée pour un mois d’hiver (janvier) et un mois d’été (juillet) en utilisant le modèle de transport CHIMERE forcé par ECMWF et les flux de surface (EDGAR et VPRM). Le résultat de ce travail permet une réduction des incertitudes des bilans nationaux à hauteur de 35 %, et la quantification les émissions de CO2 et de CH4 à l'échelle nationale et régionale. Par contre, cette inversion ne contraint que partiellement les flux d’émissions. Cependant, la question sur l’efficacité de la quantité d’informations disponibles ressort à nouveau. / Since the industrial revolution, the economic and the demographic growths have increased exponentially,leading to an enhancement of the fossil fuels combustion, such as coal, oil, and natural gas. Consumingthese source of energy amplifies the greenhouse gas emissions, mainly carbon dioxide (CO2) and methane(CH4), whose accumulation in the atmosphere lead to the increase of the greenhouse effect. According tothe 5th assessment report of the Intergovernmental Panel on Climate Change (IPCC), it is extremely likely(95-100% of certainty) that the observed increase in the greenhouse effect is related to the increase of theanthropogenic emissions. However, the estimations of the GHG budget at the regional and the nationalscales remains highly uncertain. The aim of this thesis is to improve the estimation of the CO2 and CH4fluxes in France, using data assimilation techniques and atmospheric measurements provided by theIntegrated Carbon Observation System (ICOS) network.The first phase focuses on analyzing the measured CO2, CH4, and CO (Carbon monoxide) atmosphericconcentrations provided by surface monitoring stations. This study is concerned with the problem ofidentifying atmospheric data influenced by local emissions that can result in spikes in the GHG time series.Three methods are implemented on continuous measurements of four contrasted atmospheric sites. The aimof this analysis is to evaluate the performance of the used methods for the correctly detect the contaminateddata. This work allows us to select the most reliable method that was proposed to perform daily spikedetection in the ICOS Atmospheric Thematic Centre Quality Control (ATC-QC) software.Secondly, we simulate the atmospheric concentrations of CO2 and CH4 using the chemistry transport modelCHIMERE in a domain centered over France for the year 2014. The objective of this study is to evaluate thesensitivity of simulated concentrations using different input data (sensitivity to the meteorological transportand sensitivity to the surface fluxes). This work led to the quantification of both the transport and surfacefluxes errors based on the combination of different simulations. Thus, the most reliable combination of thebest input data was selected for the flux inversion study.Lastly, the measured CO2 and CH4 concentrations are used by the PYMAI inversion system (Berchet et al.,2013 and 2015) in order to estimate the CO2 and CH4 fluxes in France. The Inversion is performed for onemonth in winter (January) and one month in summer (July), using the transport model CHIMERE. Theinversion results have provided very interesting results for the regional estimation of the CO2 and CH4surface fluxes in France with an uncertainty reduction that may attain 35% of the national totals.

Graves¡¦ disease (GD), an organ specific autoimmune disease, requires two signals to activate the T cells. In addition to the specific binding of T cell receptor to the antigenic peptide-MHC complex, an antigen-independent costimulatory pathway reportedly require generate subsequent cytokines and cell surface molecules. This regulation of T-cell response is a highly-organized multiple step program. T cell costimulatory signals is found to regulate the magnitude and duration of various type of autoimmune diseases. This study is to test whether genetic polymorphism of these costimulatory genes is related with the disease susceptibility or progression. We anticipated that the candidate genetic makers are beneficial for importing GD management. We recruited 262 GD patients from the Outpatient Department of Endocrine and 200 healthy controls from the Health Screening Center of Chang Gung Memorial Hospital in Kaohsiung.The GD patients were divided into three groups: recurred within 9 months (n=91), between 10-36 months (n=65), and more than 36 months (n=106). Clinical and laboratory attributes included: the genotypes of CTLA-4, CD28, ICOS, PD-1 and CD40; serum levels of T4, T3 and TSH; goiter size and TSH-receptor antibodies at the beginning and end of treatment. Genomic DNA was extracted from peripheral blood leucocytes by kit. The single nuclotide polymorphisms of the candidate genes were genotyped by polymerase chain reaction- restriction fragment length polymorphism and TaqMan® SNP Genotyping Assays with specific primers. Linkage disequilibuium between pairs of polymorphism was estimated by Haploview software. Haplotype analyses were performed using the Hap-Clustering program. Variance and correlation of data was statistically analyzed by Chi-square, general liner model, multiple logistic regression analysis and Kaplan-Meier plot. A p value <0.01 was considered significant. The results showed:(1) Genetic polymorphism within the costimulatory molecules affected the susceptibility and progression of GD; (2) GD patients carried more risk alleles than the controls; (3) Within the GD group, patients harboring more risk alleles wound relapse earlier after drug withdrawal; (4) Number of risk alleles, goiter size and TBII levels at end of treatment were independent predictors of disease relapse; (5) A risk score calculation based on odds ratio of risk alleles correlated with patients¡¦ relapse time after drug withdrawal. We concluded that patients¡¦ genetic makers of costimulatory molecules may be helpful in choosing appropriate treatment for GD.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-11-22T20:32:41Z No. of bitstreams: 1 AlcineyMirandaCabral_TESE.pdf: 9055485 bytes, checksum: 069ff7e30ae747a94ed4317fcd5f7186 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-11-23T22:39:21Z (GMT) No. of bitstreams: 1 AlcineyMirandaCabral_TESE.pdf: 9055485 bytes, checksum: 069ff7e30ae747a94ed4317fcd5f7186 (MD5) / Made available in DSpace on 2017-11-23T22:39:21Z (GMT). No. of bitstreams: 1 AlcineyMirandaCabral_TESE.pdf: 9055485 bytes, checksum: 069ff7e30ae747a94ed4317fcd5f7186 (MD5) Previous issue date: 2017-06-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O desafio inovador deste trabalho foi aplicar o m?todo qu?mico de S?ntese por Rea??o de Combust?o adaptada ?s perovskitas multiferr?icas BiFeO3, nas suas formas monof?sicas. Inicialmente, foi realizado a s?ntese para o BFO nas temperaturas ap?s a rea??o (BFOP.C.), a 700 oC (antes da Temperatura de Curie, BFO700) e a 830oC (pr?xima a Temperatura de Curie, BFO830). Na segunda etapa da s?ntese, foi definida a temperatura de 700 oC no sistema de dopagens LaxBi1-xFeO3 (x= 0.00, 0.10, 0.20 e 0.30), formando BFO700, L0.10BFO700, L0.20BFO700 e L0.30BFO700. Os materiais sintetizados foram caracterizados por Refinamento de Rietveld (DRX ? Rietveld), Espectroscopia no Infravermelho M?dio com Transformada de Fourier (FTIR), Microscopia Eletr?nico de Varredura com Espectroscopia por Energia Dispersiva (MEV/EDS/MAPEAMENTO), An?lises T?rmicas (TG/DTA), Propriedades Magn?ticas e El?tricas (???,??,??? ? ??? ?). Os resultados das an?lises dos par?metros estruturais, el?tricos e magn?ticos das perovskita de BiFeO3 (BFO) nas fases monof?sicas, indicaram simetria/grupo espacial Rombo?drica/?3? (BFOP.C.), Rombo?drica/?3? (BFO700) e Tricl?nica/?1 (BFO830) nas cer?micas. As dopagens por La3+ nas amostras BFO700, L0.10BFO700, L0.20BFO700 e L0.30BFO700 proporcionam diferentes valores Magn?ticos de Satura??es e resultaram Romboedral/?3? - 0.011314 ??.[?.?.]?1, Ortorr?mbico/???? - 0.006516 ??.[?.?.]?1, Ortorr?mbico/???? - 0.011503 ??.[?.?.]?1e Ortorr?mbico/???? - 0.009509 ??.[?.?.]?1respectivamente. As an?lises por FTIR/DRX ? Rietveld evidenciaram vibra??es caracter?sticas em 530.34, 531.61 e 551.51cm-1 que atribu?dos s?o ?s perovskitas de BiFeO3 com distor??o octaedral (FeO6). O estudo das estruturas sintetizadas apontaram para as cer?micas antiferromagneticas com distor??o octaedral. Na faixa 0 ? 0.5 GHz a dieletricidade das cer?micas foi maior que 6 e a tangente de perdas menores que -0.13. O fator de qualidade da antena com as amostras cer?micas apresentaram valores entre -10 dB e -25dB. Os resultados indicaram o alto potencial do BFO e LBFO como substratos cer?micos qu?micos em antenas magnetoferr?icas. / The innovative challenge of this work was to apply the chemical synthesis method by combustion reaction adapted to multiferroic perovskite in the forms BiFeO3 single. Initially, the synthesis of the BFO in temperatures after the reaction (BFOP. C.), 700 oC (before to curie temperature, BFO700) and 830 oC (near to curie temperature, BFO830). Secondly, the synthesis was defined the temperature of 700 oC in the doping system LaxBi1-xFeO3 (x= 0.00, 0.10, 0.20 and 0.30), forming BFO700, L0.10BFO700, L0.20BFO700 and L0.30BFO700. The sintetized materials were characterized by Rietveld Refinement (DRX ? Rietveld), Medium Fourier Transform Infrared Spectroscopy (FT ? IR), Scanning Electron Microscopy and Energy Dispersive X ? ray Spectroscopy (SEM/EDX/MAPPING), Thermical Analysis (TG/DTA), Magnetic and Electrical Properties (???,??,??? and ??? ?). The analytical results of the structural parameters, eletrics and magnetics of the perovskites of BiFeO3 (BFO), in the single phases, indicated symmetry/space group Rhombohedral/?3? (BFOP.C.), Rhombohedral/?3? (BFO700) and Triclinic/?1 (BFO830) in the ceramics. The doping by La3+ in the samples BFO700, L0.10BFO700, L0.20BFO700 and L0.30BFO700 showed different magnetic saturation of materials, resulting in Rhombohedral/?3? - 0.011314??.[?.?.]?1, Orthorhombic/???? - 0.006516 ??.[?.?.]?1, Orthorhombic/???? - 0.011503 ??.[?.?.]?1and Orthorhombic/???? - 0.009509 ??.[?.?.]?1 respectively. The analysis by FT ? IR/XRD ? Rietveld showed vibration characteristics in 530.34, 531.61 and 551.51cm-1 that are attribute to perovskites of BiFeO3 with octahedral distortion (FeO6). The study of the sintetized structures pointed for the antiferromagnetic ceramics with an behaviour octaedral distortion. In the range 0 ? 0.5 GHz, the dielectricity of the ceramics was more that 6 and the smaller loss tangent that -0.13. The quality factor of the antenna with the ceramic samples presented value between -10 dB and -25dB. The results indicated the potential high of the BFO and LBFO as substrates chemical ceramics in magnetoferroic antennas.

Metal-induced autoimmunity is an experimental model of environmentally induced autoimmune syndrome. Subtoxic doses of heavy metals administered to genetically susceptible mice resulted in the production of highly specific IgG antinucleolar antibodies (ANoA) accompanied by lymphoproliferation and serum increases in IgG1 and IgE. In this study, the induction of tolerance to mercuric chloride (HgCl2)-induced autoimmunity by pre-exposure to the low-dose mercury was reported. The ultimate mechanisms through which the immune system obtains the tolerance to a low dose of heavy metals remain unknown. The previous experiment showed that CD4+CD25+ regulatory T cells (Tregs) contributed to the maintenance of immunological self-tolerance and to the prevention of autoimmune diseases. The tolerized mice had a higher percentage of Tregs and ICOS+ regulatory T cells than the nontolerized mice. ICOS (Inducible T-cell COStimulator) is a costimulatory receptor homologous to CD28 and CTLA-4. The expression of ICOS occurs on activated T cells and is dependent upon TCR and CD28 signals. The anti-ICOS blockade restored the ability of tolerized mice to produce elevated amounts of IgG1, IgE and anti-nucleolar antibodies. The ICOS expression on Tregs and Teff cells increased after the mercury challenge. Mice that received anti-ICOS had a low percentage of Tregs and showed an increased production of several cytokines. Taken together, these results suggested that Tregs maintained the immune tolerance in response to chemical challenges. The ICOS pathway is important for the differentiation of Tregs and blocking this pathway could prevent peripheral tolerance to the low dose mercury. The results also showed the splenocytes from the tolerized group produced a higher amount of IL-10 than the nontolerized group. This promoted us to study the role of IL-10 in tolerance induction. To validate the role of Interleukin-10 in tolerance maintenance, the tolerized mice were treated with blocking anti-IL-10 and anti-IL-10 receptor mAb. Those tolerized mice treated with IL-10 blocking antibodies produced a higher amount of serum IgG1, IgE and anti-nucleolar antibodies compared with the group treated with control antibodies. This suggested IL-10 is critical in maintaining the peripheral tolerance in a mercury treated mouse model. All Trans retinoic acid (ATRA) is the metabolically active derivative of vitamin A and functions as potent regulator of gene expression. Vitamin A and retinoic acid play vital roles in the homeostatic control of the immune system because vitamin A-deficient individuals are incapable of controlling bacterial, viral, and protozoan diseases. In this study, the role of ATRA was reported in the first time in mercuric chloride (HgCl2)-induced autoimmunity by feeding the ASW mice ATRA. The results showed that retinoid acid exacerbated the mercury-induced autoimmunity. To study whether retinoic acid plays a role in low dose mercury induced tolerance, three groups of tolerized ASW mice were treated with either ATRA, mercury or both. The results showed retinoic acid could break the low-dose mercury-induced tolerance. The splenocytes from the group that received both mercury and ATRA treatment produced much more IL-2 and IFN-γ. This group had the lowest percentage of IL-10+ cells. The group that received ATRA had a lower percentage of early apoptosis cells. To further study the mechanism, a PCR array that included 84 genes, involved in T cell and B cell activation, was used to study four groups of mice treated with mercury, ATRA, both or neither. 10 candidate genes were selected and analyzed by Real-Time PCR to validate the PCR array results. Further study is needed to characterize the expression and the role of molecules that are upregulated by mercury and ATRA. / Microbiology and Immunology

La réponse immune représente une réponse complexe à laquelle correspond une succession d’événements orchestrés finement. Parmi les mécanismes qui régulent la réponse immune, les lymphocytes T régulateurs (Tregs) assurent le maintien de la tolérance en périphérie et le contrôle des réponses immunes adaptatives. Ils représentent une population hétérogène et leurs mécanismes de suppression sont toujours l’objet d’intenses recherches. Suivant le contexte de suppression et leur nature, les lymphocytes Tregs réalisent une inhibition de l’activation des lymphocytes Th, soit directement, soit via la modulation de la fonction des cellules dendritiques (DC). Dans un modèle d’immunisation par des cellules dendritiques chargées de KLH, les lymphocytes Tregs naturels contrôlent sélectivement l’initiation des réponses de type Th1/CTL spécifiques de l’antigène. Le but de ce travail était de définir quels sont les acteurs potentiels du contrôle de cette réponse. A l’aide de l’anticorps PC61 dirigé contre le récepteur CD25 et éliminant les lymphocytes Tregs naturels, nous avons montré que le ligand de costimulation CD70 joue un rôle clé dans leur régulation de la réponse Th1/CTL (Article 1). Ainsi, dans des conditions normales, la cytokine IL-12 induit principalement l’initiation de la réponse Th1 in vivo, tandis qu’en l’absence de lymphocytes Tregs naturels, la voie CD70/CD27 est une voie alternative d’induction de l’IFN-γ. Cette voie d’activation pourrait être opérationnelle dans certains contextes infectieux lorsque les lymphocytes Tregs sont déstabilisés voire éliminés, par exemple lors d’infections par Toxoplasma gondii ou par les virus HTLV1, SIV ou HIV. Nous avons montré que les lymphocytes Tregs naturels diminuent l’expression du ligand CD70 sur les DC, de manière dépendante de son récepteur CD27. Ensuite, nous nous sommes intéressés à une deuxième population de lymphocytes T régulateurs, les lymphocytes Tregs ICOShigh induits in vivo par le traitement avec l’anticorps anti-CTLA-4. Dans le cadre d’une colite induite par l’agent alkylant TNBS et mettant en jeu une réponse Th1, cette population de lymphocytes Tregs amplifiée par le traitement à l’anticorps anti-CTLA-4 régule la réponse immune via la cytokine anti-inflammatoire IL-10 et l’enzyme immunosuppressive IDO (Article 2). Ainsi, les résultats obtenus nous ont permis de répondre à notre objectif et de définir certains mécanismes de suppression des lymphocytes Tregs naturels et des lymphocytes Tregs induits. Dans la dernière partie de ce travail, nous avons cherché à comparer les populations de lymphocytes Tregs naturels et ICOShigh présentes dans l’intestin d’une souris naïve. Une analyse transcriptomique a révélé que ces deux populations s’opposent sur les plans phénotypique et fonctionnel. Nous proposons un modèle dans lequel les deux populations de lymphocytes Tregs agiraient en synergie pour maintenir l’homéostasie intestinale. Les lymphocytes Tregs ICOShigh différenciés au niveau local et continuellement activés contrôleraient la réponse inflammatoire associée à la présence de la flore commensale. Les lymphocytes Tregs naturels, en quiescence dans les ganglions mésentériques, n’interviendraient qu’en cas d’infection par des pathogènes. L’étude des lymphocytes T régulateurs soulève un certain nombre de concepts clés de l’immunité : la spécificité des réponses, la distinction des microorganismes commensaux et pathogènes… Mieux connaître les lymphocytes Tregs dans un modèle murin permettra de mieux comprendre les réponses inflammatoires intestinales chroniques observées chez l'homme et d’envisager, à terme, de nouveaux traitements. / An immune response is complex and implies numerous sequential events. It is regulated by different mechanisms, among which regulatory T cells maintain peripheral tolerance and control adaptive immune responses. Regulatory T cells are very heterogeneous and suppress immune responses through different mechanisms, still under investigation. They can inhibit T cell activation directly or through the modulation of dendritic cell function, depending on their nature and the tissular context. In a dendritic cell-mediated immunization model, naturally occurring regulatory T cells selectively control the priming of antigen-specific Th1/CTL responses. Our goal was to define the potential actors of this control, targeted by natural regulatory T cells. Using the PC61 antibody which targets and depletes these cells, we showed that the costimulation ligand CD70 plays a key role in their control of Th1/CTL responses (first article). We showed that mainly IL-12 provokes Th1 development in normal conditions, wheras CD70 plays a major role in priming Th1 responses in the absence of natural Tregs. This pathway can be operational if regulatory T cells are destabilized or even depleted, for example during infection with Toxoplasma gondii or with HTLV1, SIV or HIV. We showed that natural Tregs downregulate CD70 expression on the surface of DCs. Next, we focused on another regulatory T cell population, induced in vivo by the anti-CTLA-4 mAb treatment. In a model of pro-Th1 colitis, induced by the alkylating agent TNBS, these ICOShigh regulatory T cells exert an IL-10 and IDO-dependant control over the immune response (second article). Thus, we succeeded in determining some control mechanisms of the immune response targeted by two populations of regulatory T cells. Finally, we compared two regulatory T cell populations: naturally occurring regulatory T cells and ICOShigh regulatory T cells from the intestines of naïve mice. A transcriptional analysis revealed two populations phenotypically and functionally distinct. We proposed a model in which these populations act synergistically and both maintain intestinal homeostasis. ICOShigh regulatory T cells might control commensal gut flora-specific inflammatory responses and quiescent natural regulatory T cells from mesenteric lymph nodes might control potential pathogen infections. As a conclusion, this study raises some immunological issues: specificity of immune responses, distinction between commensal and pathogenic microorganisms… A better knowledge of these regulatory populations will lead to a better understanding of human intestinal responses and in the medium term will lead to new therapeutic approaches and tools.

Submitted by Alexandre Soares (alexandredesoares@yahoo.com.br) on 2016-08-25T13:32:23Z No. of bitstreams: 1 kariny_ferreira_moreira.pdf: 704247 bytes, checksum: 3674bfe36cc3e9be9a82f1bbfa71c745 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-09-08T18:43:13Z (GMT) No. of bitstreams: 1 kariny_ferreira_moreira.pdf: 704247 bytes, checksum: 3674bfe36cc3e9be9a82f1bbfa71c745 (MD5) / Made available in DSpace on 2016-09-08T18:43:13Z (GMT). No. of bitstreams: 1 kariny_ferreira_moreira.pdf: 704247 bytes, checksum: 3674bfe36cc3e9be9a82f1bbfa71c745 (MD5) Previous issue date: 2015 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A busca por alimentos alternativos dentro da nutri??o animal tem crescido bastante em virtude da demanda por alimentos vi?veis economicamente e oferta de coprodutos agroind?strias. Assim, objetivou-se com o presente trabalho avaliar a substitui??o, em n?veis crescentes: 0, 25, 50, 75% da prote?na do concentrado pela prote?na bruta oriunda do farelo de crambe em dietas destinadas a cordeiros confinados. Durante todo per?odo experimental os animais receberam dietas contendo 50% de volumoso e 50% de concentrado. Foram utilizados 24 cordeiros machos SRD, n?o castrados, com idade m?dia inicial de quatro meses e peso vivo m?dio inicial de 17,50?3,90 kg distribu?dos em delineamento inteiramente casualizado com seis repeti??es por tratamento confinados individualmente. Os animais foram pesados para avalia??o de ganho de peso (GP), ganho m?dio di?rio (GMD), efici?ncia alimentar (EA) e convers?o alimentar (CA) havendo controle di?rio de consumo individual. Em rela??o aos fatores antinutricionais causados pelos glicosinolato coletou-se amostras de sangue a cada 15 dias para avaliar os poss?veis efeitos hep?ticos atrav?s da enzimas alanina aminotransferase (ALT) e aspartato aminotransferase (AST) e para determina??o de nitrog?nio ur?ico s?rico. Foi realizado ensaio de cinco dias em que se avaliou-se a digestibilidade aparente total atrav?s de coleta total de fezes e consumo de nutrientes. Foram realizadas coletas spot de urina para a determina??o dos derivados de purina, efici?ncia de s?ntese de prote?na microbiana, teor de nitrog?nio ur?ico da urina e balan?o de nitrog?nio. Todos os ingredientes da dieta, sobras e fezes foram submetidos ?s an?lises para quantifica??o dos componentes nutricionais. Os resultados foram submetidos ? an?lise de vari?ncia e estudo de regress?o adotando-se o n?vel de signific?ncia de 5%, utilizando-se o programa SAS. N?o foi observado efeito dos n?veis de farelo de crambe nas dietas para ganho m?dio di?rio, efici?ncia alimentar e convers?o alimentar. Houve efeito linear decrescente em kg.dia-1 para o consumo de MS, MO, PB e CNFcp. N?o houve efeito das dietas para digestibilidade aparente da MS, MO, PB, FDNcp EE, CNFcp e CHOT. Verificou-se efeito linear crescente para os valores de efici?ncia de s?ntese microbiana em gPBmic.100gNDT-1consumido. N?o houve altera??o da atividade das enzimas hep?ticas AST e ALT. Para realizar as an?lises econ?mico-financeiras foram considerados o perfil tecnol?gico, indicadores de tamanho, indicadores zoot?cnicos e indicadores econ?micos dos sistemas de produ??o associados a diferentes estrat?gias nutricionais. Foram utilizados os dados de desempenho do presente estudo sendo consideradas as estruturas de avalia??o de custos de produ??o em custo efetivo, custo operacional total, custo total, margem bruta, margem l?quida, lucro e a taxa de retorno do capital investido. Contudo estrat?gia nutricional com inclus?o de 25% de PB do farelo de crambe (25FC) proporcionou maior n?mero de ciclos de termina??o por ano culminando em maior n?mero de animais vendidos. Verificou-se que a estrat?gia nutricionaal com inclus?o de 75% de PB do farelo de crambe (75FC) foi respons?vel pelo menor custo com a alimenta??o dos animais sendo ambas as estrat?gias 25FC e 75FC as mais rent?veis apresentando maiores valores da taxa de retorno do capital investido de 13,57% e 17,84% respectivamente. Todas as estrat?gias alimentares proporcionaram margem bruta e l?quida positiva, com m?dia de R$54326,60 ao ano e R$39643,54 ao ano, respectivamente. Pode-se concluir que o farelo de crambe, oriundo da produ??o de biodiesel, pode ser utilizado como alimento alternativo prot?ico podendo substituir em at? 75% da PB do concentrado da dieta por n?o interferir nas vari?veis de desempenho produtivo e na efici?ncia de s?ntese de prote?na microbiana apesar de causar a redu??o no consumo de MS e PB. Da mesma maneira, o aumento do consumo de glicosinolato, presente do farelo de crambe n?o causou aumento nas enzimas hep?ticas AST e ALT. A utiliza??o de at? 75% de substitui??o da prote?na do concentrado oriunda do farelo de crambe na produ??o de cordeiros de corte ? economicamente vi?vel permitindo a viabilidade da atividade em curto e ou longo prazo. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Zootecnia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / The search for alternative food in animal nutrition has grown considerably due to economically viable food supply demand, and offer of agro-industries co-products. The aim of the present study was to evaluate the replacement, at increasing levels: 0, 25, 50, 75% of feed concentrate protein, for crude protein derived from the crambe meal in diets designed to lambs. Throughout the trial period, animals were fed with a diet containing 50% roughage, and 50% concentrate. They were used 24 lambs, male not castratedbwith initial age of four months, and average body weight of 17.50 ? 3.90kg, distributed in a completely randomized design, composed with six replicates per treatment confined individually.The animals were weighed for further evaluation of weight gain (WG), average daily gain (ADG), feed efficiency (EA), and feed conversion (FC) with daily control of individual consumption. Regarding antinutritional glucosinolates factors, blood samples were collected on taken every 15 days to avaluate the possible effects caused by the liver, through the enzymes measured glucosinate aspartate aminotransferase (AST), and alanine aminotransferase (ALT) for determination of serum urea nitrogen. It was carried out apparent total digestibility assay, during five consecutive days, total and individual fecal where collected. Spot urine collections, for the purine derivatives determination, microbial protein synthesis efficiency, urea nitrogen and nitrogen balance. The results were submitted to analysis of variance compared by regression adopting the significance level of 5% using the SAS program. There was no effect of crambe meal levels in the diets for daily mean gain, feed efficiency, and feed conversion. A decreasing linear effect kg day-1 in DM, OM, CP, and CNFcp intake. There was increasing linear effect for microbial synthesis efficiency values at gPBmic.100gNDT-1 intake. There was no change in liver enzyme AST and ALT activity. To conduct the economic and financial analyzes, were considered the technological profile, size indicators, zootechnical indicators, and economic indicators of different feeding strategies. production costs for Structural Assessment in cost effective, total operating cost, total cost, gross margin, net margin, profit, and the rate of return on invested capital were considered. However, nutrional strategy with the inclusion of 25% CP crambe meal (25FC) provided a greater number of production cycle per year, culminating in more animals. It was found that the nutritional strategy with inclusion of 75% CP crambe meal most cost effective, and with greater values of capital invested return rate, 13.57%, and 17.84%, respectively. All food strategies provided positive gross and net margin, avering R$ 54,326.60 per annum and R$ 39,643.54 per annum respectively. It can be concluded that crambe meal, derived from biodiesel production, can be used as an alternative food protein, and can replace up to 75% protein diet not interfer in the productive performance, and in microbial protein synthesis efficiency despite to cause a reduction in the consumption of DM, and CP. Likewise, increasing glucosinate, coming from the crambe meal didn?t cause animals no increase in liver enzymes AST and ALT within studied levels. The use of 75% replacement of the concentrated protein derived with crambe mealbran the production cutting is economically viable enabling the viability in the short or long-term activity.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-11-08T17:45:13Z No. of bitstreams: 1 texto completo.pdf: 1216476 bytes, checksum: e23724e0d124500611def5f98c83fb29 (MD5) / Made available in DSpace on 2018-11-08T17:45:13Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1216476 bytes, checksum: e23724e0d124500611def5f98c83fb29 (MD5) Previous issue date: 208-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O futebol é o esporte mais popular do planeta, sendo este o fato que contribui para que ao longo do tempo ele tenha se tornado uma grande cadeia de geração de produtos e renda ao redor do mundo. Na América do Sul, em especial no Brasil e na Argentina, existe um enorme potencial financeiro ainda a ser explorado, visto que o processo de profissionalização da gestão dos clubes é algo recente na história desses países. Dada a grande capacidade de geração de receitas a ser alcançada pelos clubes, este trabalho buscou analisar quais os fatores preponderantes no desempenho das equipes nos campeonatos brasileiro e argentino de futebol da primeira divisão no período entre 2006 a 2017, visando assim, fornecer alternativas para os clubes, de modo que estas instituições possam aperfeiçoar a tomada de decisão e dessa forma otimizar a performance em suas ligas nacionais e, consequentemente, seus resultados financeiros. Ao utilizar o modelo MQO Pooled, foi possível observar que o valor de mercado médio do elenco, a troca de treinadores e de jogos simultâneos ao campeonato nacional são fatores que impactam significativamente no aproveitamento em número de pontos ao final da competição. Diante destes resultados é possível inferir que o nível de investimento da equipe, o planejamento quanto a escolha do comando técnico e a gestão do elenco são características que interferem no desempenho final dos times nas ligas nacionais sul-americanas. / Football is the most popular sport on the planet, this being the fact that, over time, it has become a great chain of product and income generation around the world. In South America, especially in Brazil and Argentina, there is an enormous financial potential still to be explored, since the process of professionalizing club management is a recent development in the history of these countries. Given the large revenue generating capacity to be achieved by the clubs, this work sought to analyze the preponderant factors in the performance of the teams in the Brazilian and Argentine first division soccer championships in the period between 2006 and 2017, in order to provide alternatives for the clubs, so that these institutions can improve decision making and thus optimize performance in their national leagues and, consequently, its financial results. By using the Pooled MQO model, it was possible to observe that the average market value of the cast, the exchange of coaches and games simultaneous to the national championship are factors that significantly impact the number of points at the end of the competition. In view of these results, it is possible to infer that the level of investment of the team, planning as to the choice of the technical command and the management of the cast are characteristics interfere in the final performance of the teams in the South American national soccer championships.