No description available.

No description available.

Ulm, Univ. Diss., 2007.

In einer Vielzahl von Arbeiten konnten anti-inflammatorische Eigenschaften von Ethylpyruvat (EP) aufgezeigt werden. An verschiedenen Modellen der Sepsis, des hämorrhagischen Schocks, von Verbrennungsschäden, des Apoplex oder der Ischämie und Reperfusion wurde bei der Behandlung mit EP ein protektiver Effekt sowie eine verminderte Produktion von pro-inflammatorischen Zytokinen nachgewiesen. Als biochemische Grundlage wurde die Interaktion von EP mit dem Transkriptionsfaktor NF-κB identifiziert, die spezifischen Regulationsmechanismen konnten bisher allerdings nicht zufriedenstellend aufgeklärt werden. In dieser Arbeit wurde als eine neue mögliche Erklärung für die anti-inflammatorischen Eigenschaften des EP und weiterer α-oxo-Karbonsäureester die Inhibierung der Glyoxalase I (Glo-I) aufgezeigt. In vitro-Experimente zur Enzymaktivität belegten die Hemmung der Glo-I durch EP, während α-Hydroxy-Karbonsäureester wie L-Ethyllaktat (EL) keine inhibierenden Eigenschaften aufwiesen. Dennoch waren sowohl EP als auch EL und weitere Laktatester in der Lage, die LPS-induzierte Produktion von pro-inflammatorischen Zytokinen wie IL-1β, IL-6, IL-8 und TNF-α von humanen immunkompetenten Zellen zu supprimieren und die Expression von Immunrezeptoren wie HLA-DR, CD14 und CD91 zu modulieren. Somit konnten erstmals anti-inflammatorische Eigenschaften von Laktatestern nachgewiesen sowie eine Verbindung zwischen den Glyoxalase-Enzymen und dem Immunsystem etabliert werden. Diese und weitere Ergebnisse zur Einflussnahme der Karbonsäureester auf die Zellvitalität präsentieren das Glyoxalasesystem als mögliches Ziel neuer Therapiekonzepte für die Immunsuppression und bestätigen dessen Bedeutung für die Entwicklung von Anti-Tumor-Agenzien.

Orientador: Manoel Barros Bertolo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T03:37:39Z (GMT). No. of bitstreams: 1 Pires_CatarinaFernandes_D.pdf: 2601322 bytes, checksum: 1acb7f604b763cb1fd45c71455e387bf (MD5) Previous issue date: 2014 / Resumo: O presente estudo de caso-controle avaliou 74 crianças piauienses com Artrite Idiopática Juvenil (AIJ) em suas diversas formas: Oligoarticular, Sistêmica, Poliarticular Fator Reumatoide Positivo (FR+), Poliarticular Fator Reumatoide Negativo (FR-), Artrite relacionada a Entesite (ERA), Artrite Psoriásica e Artrite indeterminada, classificadas de acordo com os Critérios da Liga Internacional de Associações de Reumatologia (ILAR) e cento e um controles saudáveis pareados de acordo com idade, sexo, procedência e etnia. Os objetivos foram identificar e determinar os alelos HLA-DR e HLA-DQ em uma amostra da população infanto-juvenil piauiense com AIJ nas formas oligoarticular, sistêmica, poliarticular FR+, poliarticular FR-, artrite psoriásica, artrite relacionada a entesite e artrite indeterminada e compará-las com as frequências observadas no grupo de controle saudáveis; conhecer os alelos HLA-DR e HLA-DQ que estão associados a maior susceptibilidade e os que conferem maior proteção na população de crianças com AIJ em suas diversas formas. As tipagens dos alelos HLA-DR e HLA-DQ foram realizadas por meio da técnica de amplificação pela Reação de Cadeia da Polimerase (PCR), utilizando cadeias específicas de primers DR e DQ. O resultado da análise demostrou que, entre todas as formas de apresentação da AIJ, houve associação estatística significativa para a susceptibilidade da doença com o HLA-DRB1*10 OR 8,8 (IC 1,1 a 74,9) e HLA-DRB1*16 OR 2,8 (IC 1,1 a 7.5). Na forma oligoarticular a associação estatística significativa para a susceptibilidade ocorreu com o alelo HLA-DRB1*08 OR 4,6 (IC 1,4 a 14,6). Na forma sistêmica, essa associação aconteceu com o alelo HLA-DRB1*10 OR 22,2 (IC 1,8 a 269,5).Na forma Poliarticular FR+ a associação estatística significativa para a susceptibilidade ocorreu com os alelos DRB1*09 OR 12,1 (IC 2,2 a 66,9) e DRB1*10 OR 28,5 (IC 2,3 a 355,0). Na forma Poliarticular FR-, a associação estatística significativa para a susceptibilidade foi com o alelo DRB1*16 OR 13,4 (IC 1,6 a 111,2). A Artrite Psoriásica não apresentou nenhuma associação estatística significativa com os HLA pesquisados. O resultado da análise demostrou que, entre todas as formas de apresentação da AIJ, houve associação estatística significativa para a proteção da doença com o HLA-DRB1*03 OR 0,7 (IC 0,5 a 0,9) e, na forma sistêmica, essa associação aconteceu igualmente com DRB1*03 OR 0,7 (IC 0,6 a 0,8). As outras formas de apresentação da doença não demonstraram associação estatística significativa para a proteção com nenhum tipo da HLA pesquisado. A população estudada não apresentou nenhum caso de ERA e de Artrite indeterminada. O estudo encontrou ainda, associação para o risco entre o HLA-DQB1*03 OR = 6,06 (IC 1,3 a 27,2) e uveíte em pacientes com a forma oligoarticular da AIJ. Desta forma, concluímos que os nossos resultados apresentam tanto semelhanças em relação a susceptibilidade, quanto diferenças, principalmente quanto a proteção, nas associações tipicamente encontradas na literatura / Abstract: This case-controle valuated 74 children from Piauí with Juvenile Idiopathic Arthritis (AIJ) in its various forms: Oligoarticular, Systemic, Polyarticular Rheumatoid Factor Positive (FR +), Polyarticular Rheumatoid Factor Negative (FR-), Enthesitis-related Arthritis (ERA), Arthritis Psoriatic and Arthritis indeterminate, classified according to the Criteria of the International League of Associations for Rheumatology(ILAR), and one hundred and one healthy control smatched according to age ,sex, origin and ethnicity. The objectives were to identify and determine the HLA-DR and HLA-DQ in a sample of Piauí juvenile population subtypes JIA in oligoarticular, systemic, polyarticular RF + polyarticular RF -, psoriatic arthritis, enthesitis-related arthritis and arthritis indeterminate and compares them with the observed frequencies in the group of healthy control; know the HLA-DR and HLA-DQ alleles are associated with increased susceptibility and conferring greater protection in the population with JIA in its various forms and identify a possible relationship between the alleles HLA-DR and HLA-DQ and the most frequent and most aggressive form of the disease in the population studied. Polymerase Chain Reaction (PCR) using primers specific chains of DR and DQ performed the typing of HLA-DR and HLA-DQ using the technique of amplification. The result of the analysis demonstrate damong all cases of JIA was statistically significant association for disease susceptibility with HLA-DRB1*10 OR 8.8 (CI 1.1 to 74.9) and HLA-DRB1*16 OR 2.8 (CI 1.1 to 7.5). In oligoarticular JIA significant statistical association to susceptibility occurred with HLA-DRB1*08 allele OR 4.6 (CI 1.4 to 14.6), association systemic form happened to HLA-DRB1*10 OR 22.2 (CI 1.8 to 269.5) allele in polyarticular RF + the statistically significant association to susceptibility occurred with the DRB1*09 alleles OR 12.1(CI 2.2 to 66.9) and DRB1*10 OR 28.5(CI 2.3 to 355.0) in polyarticular RF- significant statistical association to susceptibility was with DRB1*16 allele OR 13.4 (CI 1.6 to 111.2). The Psoriatic Arthritis showed no statistically significant association with HLA surveyed. The result of the analysis demonstrate damong all cases of JIA was statistically significant association for disease protection with HLA-DRB1*03 OR 0.7(CI 0.5 to 0.9) and the systemic form this association also happened DRB1*03 OR with 0.7 (CI 0.6 to 0.8). The other forms of the disease showed no statistically significant association for protection on any type of HLA searched. The study population did not show any cases of ERA and indeterminate Arthritis. The study also found, statistically significant difference between the HLA-DQB1*03 OR 6.0 (CI 1.3 to 27.2), and uveitis in patients with oligoarticular form of JIA. Thus, we conclude that our results show both similarities in terms of susceptibility, and differences, especially regarding the protection, associations typically found in the literature / Doutorado / Medicina Interna / Doutora em Ciências Médicas

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

In der vorliegenden Arbeit wurde die Präsentationsfunktion der LEW-Ratten-MHC-Klasse-II-Moleküle RT1B und RT1D für verschiedene Super- und Peptidantigene sowie die Generierung gemischter MHC-Klasse-II-Isotypen in der LEW-Ratte untersucht. Sag sind Proteine bakterieller und viraler Herkunft und führen nach Bindung an MHC-Klasse-II-Moleküle durch Interaktion mit dem TZR-Vb-Teil zu einer von der TZR-Spezifität unabhängigen Aktivierung von T-Zellen, die bis zu 30 % der Gesamt-T-Zellpopulation eines Organismus erfassen kann. Die dadurch bedingte Mediatorenfreisetzung aus T- oder konsekutiv aktivierten Zellen ist einerseits für die Entwicklung bestimmter akuter Krankheitsbilder wie des toxischen Schocksyndroms, Gastroenteritiden u. a. verantwortlich, kann aber auch potentiell zu einer Aktivierung autoreaktiver T-Zellen und der Entstehung von Autoimmunkrankheiten beitragen. Zur Untersuchung der LEW-MHC-Klasse-II-Charakteristika wurden zunächst mittels retroviralen Gentransfers RT1B- und RT1D-Gene in L929-Zellen übertragen und die Oberflächenexpression durch die mAK Ox6 und 14-4-4S nachgewiesen. Anschließend erfolgte der Nachweis einer Sag-Präsentation durch Stimulation des LEW-Vb8.2-TZH 53/4 durch die bakteriellen Sag SEB, SEC1-3, MAS und YPM und von aus Lymphknoten isolierten LEW-T-Zellen durch SEC1, MAS und YPM, die beide mit der durch eine HLA-DR1-positive Zelllinie getragenen Aktivierung verglichen wurden. Beide Experimente ergaben für die Sag des primär humanpathogen Staph. aureus eine weitaus stärkere Reaktivität in Anwesenheit humaner gegenüber LEW-MHC-Klasse-II-Molekülen bei RT1B-dominierter Antwort innerhalb der präsentatorischen Rattenmoleküle. Für SEB ergaben sich zusätzlich Hinweise auf eine MHC-Klasse-II-unabhängige Aktivierung. Das von Yersinia pseudotuberculosis produzierte Sag YPM wurde ebenfalls wesentlich besser von humanen als LEW-MHC-Klasse-II-Molekülen präsentiert, zeigte allerdings nur geringe Unterschiede zwischen RT1B und RT1D. Für das aus Nagern isolierte Sag von Mykoplasma arthritidis MAS konnte eine präferentielle Bindung an RT1D mit HLA-DR1-ähnlicher Stimulationskapazität nachgewiesen werden. Darüber hinaus wurden die generierten Zelllinien auf Präsentation der definierten Antigene L.casein und gpMBP gegenüber reaktiven TZH getestet. Dabei konnte die RT1D-restringierte Antwort von Klon19 auf L.casein und die RT1B-restringierte Antwort von 53/4 auf gpMBP bestätigt werden. Ebenfalls wurden die erstellten Transfektanten mit einem viralen Sag der Maus, dem vSag7-Gen des mtv7, transfiziert und auf Stimulation des TZH RG17 und von LEW-Lymphozyten getestet. Dabei zeigte sich eine geringe Reaktivität gegenüber den erstellten Transfektanten, die RT1B-dominiert war. Gleichzeitig ergaben sich in der Auswertung Hinweise für einen vSag7-Transfer von MHC-Klasse-II-negativen Produzenten auf MHC-Klasse-II-positive Rattenzellen, die durch weitere Experimente inklusive eines In-vivo-Tests bestätigt werden konnten. In der Generierung gemischter Isotypen aus MHC-Klasse-II-Einzelkettengenen der LEW-Ratte konnte gezeigt werde, dass die Übertragung der RT1DaBb-Genkombination mit Hilfe eines retroviralen Gentransfers auf P80- und L929-Zellen nicht zu einer sicher detektierbaren Oberflächenexpression führte, auch nicht bei Koübertragung der invarianten Kette der Maus. Durch einen Western Blot unter reduzierenden Bedingungen konnte eine bezüglich Molekulargewicht und Quantität zu einem regulären RT1B-Molekül differente RT1Bb-Einzelkette in der Kombination RT1DaBb nachgewiesen werden. / In the work at hand the presenting function of LEW rat MHC class II molecules RT1B and RT1D for various superantigens and antigens as well as the generation of mixed MHC class II isotypes in the LEW rat have been investigated. Superantigens are proteins of bacterial and viral origin, which lead to a TCR-specificity-independent activation of up to 30 % of the individual's T-cell population by interacting with the Vb part of the T-cell receptor after having bound to an MHC class II molecule. The release of mediators by T- and consecutively activated cells causes on the one hand development of acute diseases like toxic shock syndrome, gastroenteritis and other, but can also potentially activate autoreactive T-cells and lead to autoimmune diseases. In order to examine characteristics of LEW MHC class II molecules, first RT1B and RT1D chain genes were transferred into L929 cells via a retroviral transfection system and surface expression was demonstrated by using the monoclonal antibodies Ox6 and 14-4-4S. Successively, superantigen presentation was verified by stimulation of the LEW Vb8.2+ T-cell hybridoma 53/4 by bacterial superantigens SEB, SEC1-3, MAS and YPM and of LEW T-cells isolated from lymph nodes by SEC1, MAS and YPM. Both results were compared to activation in context of an HLA-DR1+ cell line. Experiments showed a much higher reactivity for the superantigens of human pathogen staph. aureus in presence of human versus LEW MHC class II molecules and an RT1B dominated answer amongst LEW presentatory molecules. Additionally, clues for MHC class II independent activation were found in case of SEB. The superantigen of yersinia pseudotuberculosis YPM was also much better presented by human than LEW MHC class II molecules while showing little differences between RT1B and RT1D. MAS bound preferentially to RT1D and equal stimulative capacity compared to HLA-DR1 could be detected. Accessorily, generated cell lines were analysed for presentation of peptide antigens L.casein and gpMBP towards reactive T-cell hybridomas, in which RT1D-restricted answer of Klon19 to L.casein and RT1B-restricted answer of 53/4 to gpMBP could be confirmed. Also generated cell lines were transfected with a viral mouse superantigen, the vsag7 gene of mtv7, and tested for stimulation of the RG17 T-cell hybridoma and LEW lymphocytes. Low reactivity towards transfected cell lines was detected, which was dominated by RT1B. Additionally, evidence for a transfer of vsag7 from MHC class II- producers to MHC class II+ rat cells could be found, which was enhanced by additional experiments including in vivo testing. Attempting to create mixed isotypes consisting of LEW rat MHC class II chains, it was demonstrated, that transferring the gene combination RT1DaBb via retroviral gene transfer into P80 and L929 cell lines resulted in no certain surface expression, also not under cotransfection of these cells with mouse invariant chain. Using western blot method under reducing conditions, a RT1Bb single chain different to the one of the regular RT1B molecule concerning molecular weight and quantitiy could be detected in the combination RT1DaBb.

Thèse (M.Sc.) -- Université Laval, 1997. / Bibliogr.: f. 60-74. Publ. aussi en version électronique.

Thèse (M.Sc.) -- Université Laval, 1997. / Les chap. 2 à 5 rédigés par l'auteur et collab. en anglais, avec résumés en français, ont été soumis ou publiés dans différents titres de périodiques. Bibliogr.: f. 179-191. Publié aussi en version électronique.

Made available in DSpace on 2016-04-04T12:45:12Z (GMT). No. of bitstreams: 2 nathalia_sa_ioc_mest_2015.pdf: 3537847 bytes, checksum: 61867e9568232ee7e431513765bfe8c5 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / Atualmente, a tuberculose (TB) e a síndrome da imunodeficiência humana (HIV) são as duas principais doenças infecciosas que levam à óbito no mundo. A infecção pelo HIV aumenta o risco de adoecimento por TB, sendo essa uma das mais frequentes doenças oportunistas. Em certos pacientes com tuberculose e infectados pelo HIV-1 que recebem tratamento para os dois agravos, uma profunda reação patológica inflamatória pode surgir, causando um efeito contrário ao esperado. Esse quadro patológico paradoxal é denominado IRIS (Síndrome Inflamatória da Reconstituição Imune). Os fatores associados ao risco da IRIS ainda não estão completamente compreendidos. Estudos sobre a patogênese desta síndrome relatam que tanto a combinação da carga antigênica quanto a susceptibilidade genética do hospedeiro podem influenciar o aparecimento da síndrome. No presente estudo, avaliamos a distribuição e o impacto dos genótipos HLA-B, HLA-DRB1 e KIR em indivíduos com tuberculose infectados pelo HIV-1, além do papel desses genes na ocorrência da IRIS. O estudo é retrospectivo, e incluiu 61 pacientes acompanhados no período de 2006 a 2012 no âmbito do projeto \2018\2019Síndrome de reconstituição imune: avaliação da resposta imune em pacientes com tuberculose em uso de HAART\2019\2019, conduzido em colaboração com o Instituto Nacional de Infectologia (INI/FIOCRUZ) Os dados das frequências gênicas dos pacientes foram comparados com dados disponíveis para a população brasileira. Os alelos HLA-B mais frequentes foram: B*15; B*44; B*35 e B*07, enquanto que os alelos HLA-DRB1 mais frequentes no estudo foram: DRB1*07, DRB1*11, DRB1*04 e DRB1*15. Esses resultados corroboram com estudos prévios da população Brasileira e, apesar de terem sido observadas algumas diferenças nas frequências alélicas entre os grupos com IRIS e sem IRIS, estas não atingiram significância estatística. Uma tendência à significância envolvendo o alelo HLA-B*42 foi observada entre os grupos IRIS x não IRIS (p= 0,064). Com relação às frequências dos genes KIR, estas foram semelhantes às descritas para a população Brasileira, porém não houve diferenças estatisticamente significativas relativas à distribuição das frequências dos diferentes genótipos KIR e seus haplótipos quando se comparou o grupo de pacientes com IRIS versus sem IRIS. Portanto, com base nestes achados, não foi possível inferir associações entre estes marcadores genéticos e a ocorrência de IRIS. Contudo, esse trabalho foi pioneiro na descrição da distribuição dos alelos HLA-B, HLA-DRB1 e KIR em indivíduos com tuberculose infectados pelo HIV-1 que, no seu conjunto, visam contribuir para a discussão sobre o impacto de genes do hospedeiro no contexto dos dois agravos estudados e na ocorrência da IRIS / Currently, tuberculosis (TB) and human immunodeficiency syndrome (HIV) are the two major infectious diseases that lead to death in the world. HIV infection increases the risk of TB illness, being one of the most frequent opportunistic diseases. In some patients with tuberculosis and HIV - 1 that received treatment for the two diseases, a deep pathological inflammatory reaction can ar ise, causing an effect contrary to the expected. This paradoxical pathological condition is called IRIS ( inflammatory syndrome of reconstitution immunity ) . The factors associated with the risk of IRIS are not yet completely understood. Studies on the patho genesis of this syndrome report that both the combination of antigenic load and the genetic susceptibility of the host can influence the appearance of the syndrome. In the present study, we evaluated the distribution and the impact of HLA - B, HLA - DRB1 and K IR genotypes in individuals with tuberculosis infected with HIV - 1, as well as the role of these genes in the occurrence of IRIS. This study is retrospective and included 61 patients followed up between 2006 and 2012 in the context of the project ''Immune r econstitution syndrome: evaluation of immune response in patients with tuberculosis in use of HAART’', held in collaboration with the National Infectology Institute ( INI/FIOCRUZ). The gene frequency data of patients were compared with data from the Brazili an population. HLA - B alleles more frequent were B*15; B*44; B*35 and B*07, while HLA - DRB1 alleles frequent in the study were DRB1*07, DRB1*11, DRB1*04 and DRB1*15. These results corroborate previous studies in the Brazilian population and, although some di fferences in the allele frequencies could be observed between the groups with and without IRIS, none of these was statistically significant. A trend to significance involving the allele HLA - B*42 was observed between IRIS x non - IRIS groups (p =0.064). Conce rning KIR genes frequencies , they were similar to those described for the Brazilian population, but no statistically significant difference in the distribution of KIR genotypes and haplotypes was observed in the comparison of IRIS versus non - IRIS pa tients. Therefore, based on our findings it was not possible to infer any association between these genetic markers and the occurrence of IRIS. However, this study was pioneer in describing the distribution of HLA - B, HLA - DRB1 and KIR alleles among individu als with tuberculosis infected with HIV - 1. These results contribute to the discussion of the impact of host genes in the context of the two diseases studied and in the occurrence of IRIS.