English Abstract Crohn's disease together with ulcerative colitis, is a type of inflammatory bowel disease (IBD) with increasing incidence and prevalence in developed countries. IBD is an immunologically mediated multifactorial disease and it's mechanism of action is still unknown. Current well- established treatment targets the inflammation with corticosteroids and immunosuppressive drugs. Apart from the intestinal inflammation, which is the primary target of the treatment, patients are characteristically afflicted with intestinal dysbiosis. Therefore, possible interventions might be an adjuvant or biological therapy. Adjuvant therapy directly aims the microbiota with probiotics, whereas the target of biological therapy is TNF-α, a pro- inflammatory cytokine excessively secreted by macrophages. The aim of this thesis is to evaluate intestinal microbiota composition changes in IBD patients with regard to courses of adjuvant and biological therapy. Bacterial diversity was analyzed using three different DNA extraction techniques. Rapid beat beating + column (RBB+C) was chosen for analyzing patient samples, as it showed the highest DNA yield and the highest DNA purity. Primarily the bacterial diversity was analyzed using degradation gradient gel electrophoresis (DGGE) with subsequent sequencing of bands of...

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No. of bitstreams: 1 Tese - Antonio Luiz Ribeiro Boechat Lopes.pdf: 13031715 bytes, checksum: d88fbd133c649844648684a05e01cc76 (MD5) Previous issue date: 2010-12-15 / Não Informada / Rheumatoid Arthritis (RA) is a autoimmune disease that affects 1% of worldwide population and promotes polyarthritis and joint destruction with work disability. About 60% of the risk factors of RA are related to genetic characteristics of individuals. The functional polymorphisms of the TNF gene -308 G/A andPTPN22 1858 C/T were associated with RA in several studies.The aim of this study was to investigate the influence of the TNF 308 G/A polymorphism in the promoter region of the tumor necrosis factor- gene and PTPN22 1858 C/T on rheumatoid arthritisand tuberculosis patients from the Brazilian Amazon. A total of 545 individuals 205 healthy controls without arthritis and 132 individuals suffering from rheumatoid arthritis and 208 tuberculosis patients  were genotyped for these polymorphisms using a methodology based on PCR-RFLP. Rheumatoid factor, age more than 60 years old and more than 10 disease years, was found to be risk factors for systemic disease (p=0,0001). The frequency of the A allele (TNF2) in rheumatoid arthritis sufferers was not found to be significantly higher than in the controls (p=0.671; OR=1.16; confidence interval=0.59 – 2.25). However, using a logistic regression model when the patients were stratified according to whether the manifestations were preponderantly articular or systemic, there was a strong association between the TNF2 allele and the systemic disease (p=0.001; OR=4.75; confidence interval=1.82 – 12.40) as well as the use of anti-TNF immunotherapy (p=0.021; OR 2.93; confidence interval=1.15 –7.46). On the other hand, PTPN22 1858T allele was not associated with systemicdisease (p=0,071; OR=3,17; confidence interval=0.83 – 11.73), but we found association between this allele and biologic anti-TNF immunotherapy (p=0,021; OR=4.39; confidence interval=1.08 – 17.86). Moreover, there was found no association between PTPN22 15858 C/T and rheumatoid arthritis nor tuberculosis. These results suggest that the TNF2 allele is associated with the more serious forms of the disease in individuals from the Brazilian Amazon but not with a risk for developing RA. / A Artrite Reumatoide (AR) é uma doença autoimune que afeta 1% da população geral, promovendo poliartrite e destruição articular com variados graus de incapacidade. Cerca de 60% dos fatores de risco da AR são relacionados a caracteres genéticos do indivíduo. Os polimorfismos funcionais dos genes TNF -308 G/A e PTPN22 1858 C/T foram associados à AR e à Tuberculose em diversos estudos. O objetivo deste estudo foi analisar a influência do polimorfismo da região promotora do gene do Fator de Necrose Tumoral-α, TNF -308 G/A e do gene PTPN22 1858 C/T na Artrite Reumatoide (AR) e na Tuberculose pulmonar em indivíduos procedentes do Amazonas. Para isso, foram genotipados, pela técnica baseada em PCR-RFLP para o polimorfismo TNF 308 G/A (TNF2) e PTPN22 1858T, 545 indivíduos sendo 205 controles sem Artrite, 208 pacientes com Tuberculose e 132 portadores de AR. Não foi observado aumento da frequência dos alelosTNF2ouPTPN22 1858T em portadores de Artrite Reumatoide e Tuberculose em comparação aos controles (p=0,218; p=0,376, respectivamente). Foram identificados como fatores preditivos para manifestações sistêmicas da Artrite Reumatoide: o Fator Reumatoide positivo, a idade maior que 60 anos e o tempo de doença (p=0001).Quando os dados foram estratificados segundo as formas predominantemente articulares ou sistêmicas, o alelo TNF2 esteve fortemente associado às formas sistêmicas (p=0,001; OR=4,75; Intervalo de confiança = 1,82 – 12,40), além de estar associado ao uso de imunobiológicos Anti-TNF (p=0,021; OR=2,93; Intervalo de confiança=1,15 – 7,49). O alelo PTPN22 1858T também está associado ao uso de imunobiológicos (p=0,021; OR= 4,39; Intervalo de Confiança=1,08 – 17,86), mas não está relacionado às formas sistêmicas (p=0,071; OR = 3,17; Intervalo de Confiança = 0,83 – 11,73). Entretanto, o polimorfismo de PTPN22 1858 C/T não foi associado à Artrite Reumatoide ou à Tuberculose Pulmonar. Estes resultados sugerem que o alelo TNF2 está associado às formas mais graves da AR em indivíduos do Amazonas, mas não ao risco de desenvolver Artrite Reumatoide.