Os alelos HLA-DRB1, que codificam uma sequência de aminoácidos (QKRAA/QRRAA/RRRAA) nas posições 70 a 74 da terceira região hipervariável da cadeia &#61538;1 do gene DRB1, denominada epítopo compartilhado (EC), estão associados com maior susceptibilidade e gravidade para artrite reumatóide (AR) em diversas populações. Uma nova classificação proposta por Du Montcel et al tem sido desenvolvida para apurar a associação entre HLA-DRB1 e AR. Este estudo foi desenhado com o objetivo de determinar a frequência dos alelos HLA-DRB1 em pacientes brasileiros com AR, e sua associação com o fator reumatoide (FR), anticorpos antipeptídeos citrulinados (ACPA) e lesão radiográfica articular e óssea. Quatrocentos e doze pacientes com AR e 215 controles foram incluídos. A tipificação HLA-DRB1 foi realizada pela reação em cadeia de polimerase (PCR) usando primers específicos e hibridação com oligonucleotídeos de sequência específica (SSOP). A pesquisa de ACPA foi determinada pela técnica de ELISA e a do FR por nefelometria, a avaliação radiográfica realizada pelo método do índice de Sharp modificado de Van Der Heijde. Para análises estatísticas foram utilizados os testes do qui-quadrado, t de Student e a regressão logística. Nos pacientes com AR alelos HLA-DRB1*04:01, *04:04, *04:05 se associaram com AR (p<0,05), embora o amplo intervalo de confiança, vale a pena ressaltar a associação observada com o alelo DRB1*09:01 e a doença (p<0,05). Alelos HLA-DRB1 EC+ foram observados em 62,8% dos pacientes e em 31,1% do grupo controle (OR 3,62; p <0,001) e estiveram associados com ACPA (OR 2,03; p<0,001). Alelos DRB1 DERAA mostraram efeito protetor para a AR (OR 0,42; p<0,001). A análise da nova classificação de HLA-DRB1 mostra que S2 e S3P se associaram a AR (p<0,05). Alelos S2 e/ou S3P esteve presente em 65% dos pacientes e 32% do grupo controle (OR 3,86; p<0,001) e estiveram associados a ACPA (OR.2,11; p=0,001). Alelos S3D, S1, X mostraram efeito protetor para a AR. Os resultados obtidos neste estudo demonstram que pacientes brasileiros com AR de etnia majoritariamente mestiça, alelos HLA-DRB1 avaliados segundo a hipótese do EC e a classificação proposta por Du Montcel estiveram associados à suscetibilidade à doença e à presença de ACPA. / HLA-DRB1 alleles that encode an amino acid sequence at positions 70-74 of the third hypervariable region of the B chain of the DRB1 gene, called shared epitope (SE), are associated with increased susceptibility and severity to rheumatoid arthritis (RA) in different populations. A new classification proposed by Du Montcel et al has been developed to determine the association between HLA-DRB1 and RA. This study was designer to determine the frequency of HLA-DRB1 alleles in Brazilian patients with RA, and its association with rheumatoid factor (RF), citrullinated peptide antibodies (ACPA) and radiographic joint damage and bone. Four hundred and twelve patients with RA and 215 controls were included. The HLA-DRB1 typing was performed by polymerase chain reaction (PCR) using specific primers and hybridization with sequence specific oligonucleotides (SSOP). The survey of ACPA was determined by ELISA and the RF by nephelometry, the radiographic evaluation by index method modified Sharp Van Der Heijde. For statistical analysis we used the chi-square, Student and logistic regression. In patients with rheumatoid arthritis HLA-DRB1*04:01, *04:04, *04:05 are associated with RA (p<0,05), although the wide confidence interval, it is worth noting the association observed with the DRB1*09:01 allele and the disease (p<0,05). HLA-DRB1 SE+ were observed in 62.8% of patients and in 31.1% of the control group (OR 3.62, p<0,001) and were associated with ACPA (OR 2.03, p<0,001). DERRA alleles showed a protective effect against RA (OR 0,42, p<0,001). The analysis of the new classification of HLA-DRB1 shows that S2 and S3P were associated with RA (p<0,05). Alleles S2 and/or S3P was present in 65% and 32% of patients in the control group (OR 3,86, p<0,001) and were associated with ACPA (OR 2.11, p=0,001). S3D alleles, S1, X showed a protective effect against RA. The results of this study demonstrate that Brazilian patients with RA from mostly mixed ethnicity, HLA-DRB1 evaluated under the hypothesis of the SE and the classification proposed by Du Montcel et al were associated with disease susceptibility and the presence of ACPA.

Os alelos HLA-DRB1, que codificam uma sequência de aminoácidos (QKRAA/QRRAA/RRRAA) nas posições 70 a 74 da terceira região hipervariável da cadeia &#61538;1 do gene DRB1, denominada epítopo compartilhado (EC), estão associados com maior susceptibilidade e gravidade para artrite reumatóide (AR) em diversas populações. Uma nova classificação proposta por Du Montcel et al tem sido desenvolvida para apurar a associação entre HLA-DRB1 e AR. Este estudo foi desenhado com o objetivo de determinar a frequência dos alelos HLA-DRB1 em pacientes brasileiros com AR, e sua associação com o fator reumatoide (FR), anticorpos antipeptídeos citrulinados (ACPA) e lesão radiográfica articular e óssea. Quatrocentos e doze pacientes com AR e 215 controles foram incluídos. A tipificação HLA-DRB1 foi realizada pela reação em cadeia de polimerase (PCR) usando primers específicos e hibridação com oligonucleotídeos de sequência específica (SSOP). A pesquisa de ACPA foi determinada pela técnica de ELISA e a do FR por nefelometria, a avaliação radiográfica realizada pelo método do índice de Sharp modificado de Van Der Heijde. Para análises estatísticas foram utilizados os testes do qui-quadrado, t de Student e a regressão logística. Nos pacientes com AR alelos HLA-DRB1*04:01, *04:04, *04:05 se associaram com AR (p<0,05), embora o amplo intervalo de confiança, vale a pena ressaltar a associação observada com o alelo DRB1*09:01 e a doença (p<0,05). Alelos HLA-DRB1 EC+ foram observados em 62,8% dos pacientes e em 31,1% do grupo controle (OR 3,62; p <0,001) e estiveram associados com ACPA (OR 2,03; p<0,001). Alelos DRB1 DERAA mostraram efeito protetor para a AR (OR 0,42; p<0,001). A análise da nova classificação de HLA-DRB1 mostra que S2 e S3P se associaram a AR (p<0,05). Alelos S2 e/ou S3P esteve presente em 65% dos pacientes e 32% do grupo controle (OR 3,86; p<0,001) e estiveram associados a ACPA (OR.2,11; p=0,001). Alelos S3D, S1, X mostraram efeito protetor para a AR. Os resultados obtidos neste estudo demonstram que pacientes brasileiros com AR de etnia majoritariamente mestiça, alelos HLA-DRB1 avaliados segundo a hipótese do EC e a classificação proposta por Du Montcel estiveram associados à suscetibilidade à doença e à presença de ACPA. / HLA-DRB1 alleles that encode an amino acid sequence at positions 70-74 of the third hypervariable region of the B chain of the DRB1 gene, called shared epitope (SE), are associated with increased susceptibility and severity to rheumatoid arthritis (RA) in different populations. A new classification proposed by Du Montcel et al has been developed to determine the association between HLA-DRB1 and RA. This study was designer to determine the frequency of HLA-DRB1 alleles in Brazilian patients with RA, and its association with rheumatoid factor (RF), citrullinated peptide antibodies (ACPA) and radiographic joint damage and bone. Four hundred and twelve patients with RA and 215 controls were included. The HLA-DRB1 typing was performed by polymerase chain reaction (PCR) using specific primers and hybridization with sequence specific oligonucleotides (SSOP). The survey of ACPA was determined by ELISA and the RF by nephelometry, the radiographic evaluation by index method modified Sharp Van Der Heijde. For statistical analysis we used the chi-square, Student and logistic regression. In patients with rheumatoid arthritis HLA-DRB1*04:01, *04:04, *04:05 are associated with RA (p<0,05), although the wide confidence interval, it is worth noting the association observed with the DRB1*09:01 allele and the disease (p<0,05). HLA-DRB1 SE+ were observed in 62.8% of patients and in 31.1% of the control group (OR 3.62, p<0,001) and were associated with ACPA (OR 2.03, p<0,001). DERRA alleles showed a protective effect against RA (OR 0,42, p<0,001). The analysis of the new classification of HLA-DRB1 shows that S2 and S3P were associated with RA (p<0,05). Alleles S2 and/or S3P was present in 65% and 32% of patients in the control group (OR 3,86, p<0,001) and were associated with ACPA (OR 2.11, p=0,001). S3D alleles, S1, X showed a protective effect against RA. The results of this study demonstrate that Brazilian patients with RA from mostly mixed ethnicity, HLA-DRB1 evaluated under the hypothesis of the SE and the classification proposed by Du Montcel et al were associated with disease susceptibility and the presence of ACPA.

Submitted by Lúcia Brandão (lucia.elaine@live.com) on 2015-12-11T19:01:49Z No. of bitstreams: 1 Dhêmerson - Dhêmerson Souza de Lima.pdf: 2134258 bytes, checksum: 027d2a19329c1030eb634da31db37ae6 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-01-19T14:57:53Z (GMT) No. of bitstreams: 1 Dhêmerson - Dhêmerson Souza de Lima.pdf: 2134258 bytes, checksum: 027d2a19329c1030eb634da31db37ae6 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-01-19T15:02:05Z (GMT) No. of bitstreams: 1 Dhêmerson - Dhêmerson Souza de Lima.pdf: 2134258 bytes, checksum: 027d2a19329c1030eb634da31db37ae6 (MD5) / Made available in DSpace on 2016-01-19T15:02:05Z (GMT). No. of bitstreams: 1 Dhêmerson - Dhêmerson Souza de Lima.pdf: 2134258 bytes, checksum: 027d2a19329c1030eb634da31db37ae6 (MD5) Previous issue date: 2015-05-08 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Tuberculosis (TB) is a serious public health problem, considered as a priority by the government of Brazil since 2003. This disease is endemic in low and middle-income countries, mainly affecting the population living in urban peripheries when associated with poor living conditions, malnutrition and drug abuse. Brazil is one of 22 countries contributing to the global high TB burden; it is on 16th position in number of TB cases. In 2014, Amazonas state had the highest national incidence rate of TB (68.4 per 100.000 inhabitants). Poverty, social exclusion, operational difficulties for diagnosis and treatment of TB are crucial to maintaining the high rate of incidence. In addition, the host immunogenic factors are associated with TB. Human Leukocyte Antigen (HLA) genes are associated with susceptibility or resistance to TB. This study verifies the frequency of HLA-DRB1*04 allele and subtypes in 622 subjects (316 patients with pulmonary TB and 306 controls). HLA-DRB1*04 was more frequent in TB patients (187/316; 59,2%) than control group (101/306; 33,0%). In this study, nine subtypes of HLA-DRB1*04 were identified. The subtype HLA-DRB1*04:11:01 was associated with susceptibility to pulmonary TB (p = 0.0019; OR = 2.23; 95% CI = 1.34 to 3.70), while HLA-DRB1*04:07:01 was associated with protection (p < 0.0001; OR = 0.02; 95% CI = 0.001 to 0.33) and HLA-DRB1*04:92 was associated with transmission to pulmonary TB disease (p = 0.0112; OR = 8.62; 95% CI = 1.63 to 45.5). These results suggest three subtypes of HLA-DRB1*04 as potential immunogenetic markers in TB and can help in better understanding of mechanisms involved in disease, as well as the reasons for high rates of TB incidence in Amazonas state. / A tuberculose (TB) é um grave problema de saúde pública, considerada como prioridade pelo governo do Brasil desde 2003. A doença é endêmica nos países de baixa e média renda, acometendo principalmente a população que reside nas periferias urbanas (favelas e invasões), estando associada às más condições de moradia, à alimentação, ao saneamento básico e ao uso de drogas. O Brasil é um dos 22 países que contribuem com as maiores cargas de TB no mundo, ocupando a 16.a posição em número absoluto de casos. O Estado do Amazonas, em 2014, apresentou o maior coeficiente de incidência nacional de casos de TB (68,4 por 100 mil habitantes). Miséria, exclusão social, dificuldades operacionais de diagnósticos e tratamento da TB são preponderantes para manutenção da alta taxa de incidência. Além disso, os fatores imunogenéticos do hospedeiro são descritos e associados à suscetibilidade ou à resistência da doença. Dentre os genes mais fortemente associados à TB, estão os do Antígeno Leucocitário Humano (HLA). No presente estudo, foi investigada a frequência do alelo HLA-DRB1*04 em 622 indivíduos, 316 pacientes com TB pulmonar e 306 controles. O HLA-DRB1*04 foi frequente nos pacientes (187/316; 59,2%), quando comparado aos dados dos controles (101/306; 33,0%). Neste estudo, foram identificados nove subtipos de HLADRB1*04; destes, o HLA-DRB1*04:11:01 foi associado à suscetibilidade (p = 0,0019; OR =2,23; IC 95% = 1,34 – 3,70) e à transmissão de TB pulmonar, enquanto o HLA-DRB1*04:07:01 foi associado à proteção (p < 0,0001; OR = 0,02; IC 95% = 0,001 – 0,33), e o HLA-DRB1*04:92 foi associado à transmissão da doença (p = 0,0112; OR = 8,62; IC 95% = 1,63 – 45,5). Esses resultados sugerem três subtipos do HLA-DRB1*04 como potenciais marcadores imunogenéticos na TB, os quais podem ajudar na compreensão dos mecanismos envolvidos na doença, bem como esclarecer os motivos das altas taxas de incidência de TB no Estado do Amazonas.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23773号 / 医博第4819号 / 新制||医||1057(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 金子 新, 教授 杉田 昌彦, 教授 松田 秀一 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Kidney failure is an important cause of morbidity and mortality, frequently associated with chronic inflammation of glomeruli and immune hypersensitivity reactions. In this study we aimed to determine if there was an association between HLA alleles and end stage kidney diseases. Towards this objective, we analyzed 87 patients with an average age of 51 years and a mean of 4.5 years of hemodialysis. The main clinical diagnosis of kidney failure in this group was hypertension (38%), diabetes (25%) and glomerulopathies (23%). As a control, we utilized the HLA typing data of 17,541 voluntary marrow donors from the Brazilian national registry. HLA typing was determined by SSO kits (One Lambda, Inc.) and flow cytometry (Luminex technology). Our results demonstrated that, when compared to the normal population, there was a significant association of: 1) hypertension with HLA-A*23 (p=0.014), HLA-A*30 (p<0.001) and HLA-B*41 (p=0.016); 2) diabetes with HLA-A*23 (p=0.004), HLA-A*30 (p=0.033), HLA-B*41 (p=0.023), HLA-B*81 (p=0.020), HLADRB 1*1 (p=0.030) and HLA-DRB1*3 (p=0.013); and 3) glomerulopathies with HLA-A*32 (p<0.001), HLA-B*13 (p<0.001), HLA-B*14 (p=0.004), HLA-DRB1*4 (p=0.030), HLADRB 1*11 (p=0.008) and HLA-DRB1*15 (p<0.001). There was an association between allele frequency and protection against kidney disease in the following situations: 1) hypertension with HLA-A*3 and HLA-DRB1*4; 2) diabetes with HLA-DRB1*11; 3) glomerulopathies with HLA-DRB1*1 and HLA-DRB1*13. In conclusion, HLA alleles may be an important marker of prognosis in chronic renal disease. / A insuficiência renal, frequentemente associada com inflamação glomerular crônica e com reações de hipersensibilidade, é importante causa de morbidade e mortalidade. O objetivo deste estudo foi determinar se havia associação entre alelos HLA e doença renal crônica terminal. Foram analisados 87 pacientes com idade média de 51 anos e realizando hemodiálise há, em média, 4,5 anos. Os principais diagnósticos clínicos da insuficiência renal neste grupo foram hipertensão (38%), diabetes (25%) e glomerulopatias (23%). Como controle foram utilizados dados de tipagens de HLA de 17.541 doadores voluntários de medula óssea do registro nacional Brasileiro. A tipagem de HLA foi determinada através de kits SSO (One Lambda, Inc) e citometria de fluxo (tecnologia Luminex). Nossos resultados demonstraram que, quando comparado com a população normal, havia associação significativa de: 1) hipertensão com HLA-A*23 (p=0,014), HLA-A*30 (p<0,001) e HLAB* 41 (p=0,016); 2) diabetes com HLA-A*23 (p=0,004), HLA-A*30 (p=0,033), HLA-B*41 (p=0,023), HLA-B*81 (p=0,020), HLA-DRB1*1 (p=0,030) e HLA-DRB1*3 (p=0,013); 3) glomerulopatias com HLA-A*32 (p<0,001), HLA-B*13 (p<0,001), HLA-B*14 (p=0,004), HLA-DRB1*4 (p=0,030), HLA-DRB1*11 (p=0,008) e HLA-DRB1*15 (p<0,001). Houve associação entre a frequência de alelos e proteção contra doença renal nas seguintes situações: 1) hipertensão com HLA-A*3 e HLA-DRB1*4; 2) diabetes com HLA-DRB1*11; 3) glomerulopatias com HLA-DRB1*1 e HLA-DRB1*13. Conclusão: alelos HLA podem ser importantes marcadores do prognóstico em doença renal crônica. / Doutor em Imunologia e Parasitologia Aplicadas

Indiana University-Purdue University Indianapolis (IUPUI) / Background: Bacterial adherence to the acquired dental pellicle, important in caries, is mediated by receptor-adhesin interactions such as Streptococcus mutans antigen I/II (I/II). Ten I/II epitopes from the A, V, P and C regions were chosen to determine their reactivity in human saliva. Underlying the body’s ability to immunologically respond to bacteria that lead to caries are the human leukocyte antigen (HLA) genes, specifically HLA class II (HLA-II) genes that control antigen presentation. Previous studies suggested that a specific HLA biomarker group (HLA-DRB1*04) may have differential control of immune responses to I/II. However, it was not known whether secretory IgA (SIgA) responses to the selected epitopes from HLA-DRB1*04 positive subjects were different compared to their non-biomarker counterparts (negative), or across other caries factors, since no study to date had thus assessed these questions. Methods: Per IRB approval, the study population was divided into age, sex and race matched DRB1*04 positive (n=16) and negative groups (n=16). SIgA-epitope (and whole cell) reactivity was determined using ELISA. Other caries factors were measured. Subjects received a clinical exam by a trained examiner. ix Differences between DRB1*04 positive and negative groups were examined using a two-sided, two-sample t-test. Results: DRB1*04 positive subjects had numerically, but not statistically, higher reactivity to 9 out of 10 epitopes, the exception being residues 834-853 from the V and P regions of I/II across multiple measures. Though statistically insignificant, DRB1*04 positive subjects also exhibited 25-30 μg mL-1 less total IgA (TIgA) than negative counterparts. All clinical caries data proved inconclusive when comparing groups, likely due to exogenous factors and sample size. Conclusion: DRB1*04 positive subjects showed a trend toward lower TIgA. Moreover, they also showed a lower SIgA response across multiple measures to 834-853, the I/II V and P region epitope. This region forms a sort of functional epicenter involved in collaboration between domains along the entire I/II antigen, and governs the region involved in initial attachment to the acquired dental pellicle. This region may be involved in an in vivo discontinuous conformationally specific immunogenic epitope that serves as an HLA-II binding motif which remains elusive.