Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 172-200).

Thesis (Ph.D.)--Baylor University, 2009. / Includes bibliographical references (p. 126-143).

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20757号 / 医博第4287号 / 新制||医||1024(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 生田 宏一, 教授 濵﨑 洋子, 教授 髙折 晃史 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Background: The major histocompatibility complex (MHC) molecules play an important role in the immune response regulation and in the maintenance of the immune homeostasis. Regulation of their expression is therefore a key factor influencing the adaptive immune response. DNA methylation of gene regulatory regions is one of the mechanisms of gene expression control that affects the accessibility of DNA to transcription factors. Ageing is connected with changes in DNA methylation and increased predisposition to autoimmune diseases in older age could be associated with changes in MHC class II genes methylation. Aims: The aim of this diploma thesis is to analyze the methylation profile of DQA1 and DQB1 genes regulatory regions and to compare its differences between the generations and between individual alleles. The next aim is to compare DQA1 mRNA expression between the generations and between single alleles. Methods: DNA and RNA were isolated from blood of three age group donors. DNA was converted by the bisulfite treatment and regulatory regions of HLA class II genes were amplified and cloned into bacteria. Positive clones were sequenced and then analyzed. RNA was reverse transcribed and its expression level was determined by real-time PCR. Results: Statistically significant differences were found by...

<p>Human Papillomavirus (HPV) is a key factor in the development of cervical cancer. Out of the more than 100 known HPV types 13 are considered oncogenic. In addition to presence of the virus several other factors have been proposed to influence risk of cervical cancer. This thesis focuses on viral load and HLA class II alleles as risk factors for cervical cancer.</p><p>To enable quantification of the most common oncogenic HPV types, a real-time PCR-based assay was developed and evaluated in terms of technical sensitivity and specificity.</p><p>This assay was then employed on archival smears from 457 cases and 552 controls to assess associations between viral load and cervical carcinoma <i>in situ</i> (CIS). Whereas the data indicate a pronounced dose dependent effect of HPV 16 load on the risk of CIS, other HPV types only seem to increase CIS risk at higher viral loads. These effects were observed even when cytology indicated that cells were normal.</p><p>We then investigated viral load as a risk factor for invasive cervical carcinoma (ICC) in a retrospective study comprising 139 cases and 550 controls. Viral load contributed similarly to the risk of ICC as to the risk of CIS.</p><p>Finally, associations between HLA class II alleles, viral load and CIS were investigated. Carriers of the DRB1*1301 allele were less prone to infections and high viral loads of HPV 31 and -18/45. Moreover, DRB1*1301 had a protective effect against CIS among women infected by HPV 31 or -18/45. In contrast, carriers of DRB1*1501 and DQB1*0602 were more susceptible to infections and high viral loads of HPV 16.</p><p>These results indicate that HPV load may have HPV-type specific effects on cervical cancer risk. Furthermore, HLA class II alleles may confer either susceptibility or protection against cervical cancer by acting on the HPV infections preceding tumor development.</p>

Human Papillomavirus (HPV) is a key factor in the development of cervical cancer. Out of the more than 100 known HPV types 13 are considered oncogenic. In addition to presence of the virus several other factors have been proposed to influence risk of cervical cancer. This thesis focuses on viral load and HLA class II alleles as risk factors for cervical cancer. To enable quantification of the most common oncogenic HPV types, a real-time PCR-based assay was developed and evaluated in terms of technical sensitivity and specificity. This assay was then employed on archival smears from 457 cases and 552 controls to assess associations between viral load and cervical carcinoma in situ (CIS). Whereas the data indicate a pronounced dose dependent effect of HPV 16 load on the risk of CIS, other HPV types only seem to increase CIS risk at higher viral loads. These effects were observed even when cytology indicated that cells were normal. We then investigated viral load as a risk factor for invasive cervical carcinoma (ICC) in a retrospective study comprising 139 cases and 550 controls. Viral load contributed similarly to the risk of ICC as to the risk of CIS. Finally, associations between HLA class II alleles, viral load and CIS were investigated. Carriers of the DRB1*1301 allele were less prone to infections and high viral loads of HPV 31 and -18/45. Moreover, DRB1*1301 had a protective effect against CIS among women infected by HPV 31 or -18/45. In contrast, carriers of DRB1*1501 and DQB1*0602 were more susceptible to infections and high viral loads of HPV 16. These results indicate that HPV load may have HPV-type specific effects on cervical cancer risk. Furthermore, HLA class II alleles may confer either susceptibility or protection against cervical cancer by acting on the HPV infections preceding tumor development.

Thesis (M.Sc.)--Memorial University of Newfoundland, 2002. / Restricted until June 2003. Bibliography: leaves 189-214.

Made available in DSpace on 2014-06-11T19:32:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-11Bitstream added on 2014-06-13T19:21:45Z : No. of bitstreams: 1 melo_lms_dr_sjrp.pdf: 6146654 bytes, checksum: 861736c4cd6f9884be922b051977e3fb (MD5) / O presente estudo avaliou a resposta de anticorpos contra diferentes antígenos de merozoíto e esporozoíto de Plasmodium vivax, relacionando com as variantes da porção repetitiva do domínio central do gene da Proteína Circunsporozoítica (CSP) do parasito (VK210, VK247 e P. vivax-like) e com os polimorfismos do HLA-DRB1 no hospedeiro humano. A resposta de anticorpos foi avaliada para peptídeos das regiões conservadas e centrais variáveis da CSP, da porção N-terminal da Proteína de Superfície do Merozoíto 1-MSP1 (Pv200L), e recombinante do Antígeno 1 de Membrana Apical (AMA-1) e a Proteína de ligação ao Duffy (DBP) por ELISA, em amostras de plasma de pacientes naturalmente infectados com P. vivax. Inicialmente nós avaliamos a distribuição destas variantes da CSP em cinco diferentes áreas da Amazônia a fim de entender sua atual dinâmica de transmissão. A variante VK210 continua sendo a mais prevalente em todas as áreas estudadas. No entanto, pela primeira vez documentamos a presença das variantes VK247 e P. vivax-like como infecções simples na Amazônia brasileira evidenciando um novo perfil distribuição destas, o que possa sugerir um processo de adaptação das mesmas. Quando comparamos a resposta de anticorpos e a infecção pelas variantes de P. vivax, não foram observadas associações significativas entre a presença de determinada variante da CSP e a freqüência de resposta de anticorpos contra os três peptídeos do merozoíto analisados, MSP1 (Pv200L), AMA-1 e DBP e nem contra as frações conservadas da CSP no esporozíto, N-terminal [N] e C-terminal [C]. A falta de associações significativas entre resposta sorológica contra esses peptídeos fornece informações promissoras quanto à utilização destes antígenos para o desenvolvimento de uma vacina contra malária. Todavia, a variação na porção central da CSP deve ser considerada... / The present study evaluated the antibody response against merozoite and sporozoite antigens of Plasmodium vivax and its relationship with the variants of the repetitive central region of the gene for Circunsporozoite protein (CSP) in parasite (VK210, VK247 and P. vivax-like) and, with the HLA-DRB1 polymorphisms in human host. The antibody response to synthetic peptides of the CSP conserved and variable regions and of the N-terminal portion of Merozoite surface protein - MSP1 (Pv200L), and, to recombinants peptides of the Apical Membrane Antige 1 (AMA-1) and of the Duffy Binding Protein (DBP) was evaluable by ELISA in plasma samples of malaria patients naturally infected with P. vivax. Firstly, we evaluated the CSP variants distribution among five different areas from Brazilian Amazon, in order to understand their current dynamic of transmissions. VK210 variant remains the most prevalent in all study areas. However, it is the first detection of VK247 e P. vivax-like variants as simple infection in the Brazilian Amazon, showing a new distribution profile, which may suggest an adaptation process of them. When comparing the antibody response and infection by variants of P. vivax, there were no significant associations between the presence of particular CSP variant and the frequency of antibody response against all three merozoite peptides analyzed, MSP1 (Pv200L), AMA-1, DBP and against the CSP conserved fractions in the sporozoite, N-terminal and C-terminal. The lack of significant associations among immune response against these peptides provides promising information regarding the use of these antigens for malaria vaccine development. On the other hand, the central variability of CSP should be considered to employment of this region as an immunogen, since the antibody response appears to be variant-specific. In order to evaluate the polymorphisms... (Complete abstract click electronic access below)

Background: Type 1 diabetes mellitus is a multifactorial disease caused by beta cell destruction of Langerhans pancreatic islets. From the genetic aspect the main predisposition lays on HLA class II genes (40 - 50%), molecules of which present exogenous peptides to CD4+ T lymphocytes. Enviromental factors play a crucial role in the etiopathogenesis of T1DM. Through epigenetic regulation (e.g. DNA methylation) the genetic and enviromental factors communicate. The level of methylation in the regulatory regions can significantly affect expression of these genes. Aims: The aim of the diploma thesis was to define methylation profile of HLA DQB1 alleles in type 1 diabetes mellitus patients and determine their expression. Methods: The genotyping of HLA class II genes (HLA-DRB1, HLA-DQA1, HLA-DQB1) was performed using sequence specific primers. DNA was treated with sodium bisulfite, regulatory region of HLA DQB1 was amplified and cloned into E.coli, strain DH5α/XL1-Blue. Positive clones were sent for sequencing and results analyzed. RNA was transcribed to cDNA by reverse transcription and the level of expression was analyzed by quantitative PCR. Results: Statistically significant differences were found in total methylation of DQB1*0201 and *0302 alleles in the B section of DQB1 gene. Difference in...

Orientador: Ricardo Luiz Dantas Machado / Banca: Cláudio Tadeu Daniel Ribeiro / Banca: Érika Martins Braga / Banca: Claudia Regina Bonini Domingos / Banca: Érika Cristina Pavarino Bertelli / Resumo: O presente estudo avaliou a resposta de anticorpos contra diferentes antígenos de merozoíto e esporozoíto de Plasmodium vivax, relacionando com as variantes da porção repetitiva do domínio central do gene da Proteína Circunsporozoítica (CSP) do parasito (VK210, VK247 e P. vivax-like) e com os polimorfismos do HLA-DRB1 no hospedeiro humano. A resposta de anticorpos foi avaliada para peptídeos das regiões conservadas e centrais variáveis da CSP, da porção N-terminal da Proteína de Superfície do Merozoíto 1-MSP1 (Pv200L), e recombinante do Antígeno 1 de Membrana Apical (AMA-1) e a Proteína de ligação ao Duffy (DBP) por ELISA, em amostras de plasma de pacientes naturalmente infectados com P. vivax. Inicialmente nós avaliamos a distribuição destas variantes da CSP em cinco diferentes áreas da Amazônia a fim de entender sua atual dinâmica de transmissão. A variante VK210 continua sendo a mais prevalente em todas as áreas estudadas. No entanto, pela primeira vez documentamos a presença das variantes VK247 e P. vivax-like como infecções simples na Amazônia brasileira evidenciando um novo perfil distribuição destas, o que possa sugerir um processo de adaptação das mesmas. Quando comparamos a resposta de anticorpos e a infecção pelas variantes de P. vivax, não foram observadas associações significativas entre a presença de determinada variante da CSP e a freqüência de resposta de anticorpos contra os três peptídeos do merozoíto analisados, MSP1 (Pv200L), AMA-1 e DBP e nem contra as frações conservadas da CSP no esporozíto, N-terminal [N] e C-terminal [C]. A falta de associações significativas entre resposta sorológica contra esses peptídeos fornece informações promissoras quanto à utilização destes antígenos para o desenvolvimento de uma vacina contra malária. Todavia, a variação na porção central da CSP deve ser considerada... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present study evaluated the antibody response against merozoite and sporozoite antigens of Plasmodium vivax and its relationship with the variants of the repetitive central region of the gene for Circunsporozoite protein (CSP) in parasite (VK210, VK247 and P. vivax-like) and, with the HLA-DRB1 polymorphisms in human host. The antibody response to synthetic peptides of the CSP conserved and variable regions and of the N-terminal portion of Merozoite surface protein - MSP1 (Pv200L), and, to recombinants peptides of the Apical Membrane Antige 1 (AMA-1) and of the Duffy Binding Protein (DBP) was evaluable by ELISA in plasma samples of malaria patients naturally infected with P. vivax. Firstly, we evaluated the CSP variants distribution among five different areas from Brazilian Amazon, in order to understand their current dynamic of transmissions. VK210 variant remains the most prevalent in all study areas. However, it is the first detection of VK247 e P. vivax-like variants as simple infection in the Brazilian Amazon, showing a new distribution profile, which may suggest an adaptation process of them. When comparing the antibody response and infection by variants of P. vivax, there were no significant associations between the presence of particular CSP variant and the frequency of antibody response against all three merozoite peptides analyzed, MSP1 (Pv200L), AMA-1, DBP and against the CSP conserved fractions in the sporozoite, N-terminal and C-terminal. The lack of significant associations among immune response against these peptides provides promising information regarding the use of these antigens for malaria vaccine development. On the other hand, the central variability of CSP should be considered to employment of this region as an immunogen, since the antibody response appears to be variant-specific. In order to evaluate the polymorphisms... (Complete abstract click electronic access below) / Doutor