Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-05-14T11:36:20Z No. of bitstreams: 1 468608 - Texto Parcial.pdf: 483189 bytes, checksum: b04b032c1dfe3f63ec203e3d29e68a0a (MD5) / Made available in DSpace on 2015-05-14T11:36:20Z (GMT). No. of bitstreams: 1 468608 - Texto Parcial.pdf: 483189 bytes, checksum: b04b032c1dfe3f63ec203e3d29e68a0a (MD5) Previous issue date: 2015-02-26 / Introduction: Viruses cause diseases in humans and can cause epidemics and pandemics leading to death of thousands of people. When a virus infects the body, the immune system is activated to control the infection. The primary response against viruses consist mainly of cytotoxic CD8+ T cells and antibodies produced by B cells, however, both B and CD8 + T cells require auxiliary CD4 + T cells to generate an effective response. The production of antibodies needs a specific type helper cells, CD4 + follicular T cells (TFH). Both responses can generate memory, so in a second infection by the same virus strain can respond more efficient way. In these next two articles we discuss the TFH cell function towards viral response. First we produced an article that reviews all studies that performed a connection between this cell population and different viruses, such as HIV, HBV, EBV, LCMV and influenza. We next present an original article trying to unravel the mechanism by which the Respiratory Syncytial Virus (RSV) inhibits adequate humoral immune responses. Antibodies generated in RSV infections often have low affinity and little neutralizing activity, resulting in the accumulation of inflammatory immune complexes in the lower airways of children under 2 years of age. RSV can lead to death by bronchiolitis and pneumonia in approximately 50% of cases and increases the chance of developing asthma later in life. The TFH cells play an important role in helping B cells for class switching and the production of high affinity immunoglobulin. Objective: To understand the immunological mechanisms related to TFH cells in viral responses and investigate the mechanisms underlying the effect of RSV in decreasing antibody efficiency. Methods and Results: In the first article we reviewed Medline / Pubmed for original articles published from 2008 to 2014, using the terms "TFH" and "Virus" together. In the second article was conducted original research with animal models, focused on determining the interaction of RSV with the TFH cells. In our model, RSV was able to increase the amount of TFH cells in vitro, promoting their proliferation (Ki67) and death (Annexin-V). In vivo, RSV did not induce an increase in germinal centers and decreased the amount of IgG antibodies. We observed that RSV was capable of PD-L1 induction in dendritic cells as well in B cells. Conclusion: TFH cells are crucial for anti-viral humoral responses. Different types of viruses interact through varied mechanisms with the immune system. Our results suggest that the RSV induces PD-L1 in dendritic cells and B cells, and may be able to cause death by apoptosis in TFH cells, reducing the size of germinal centers and decreasing the amount of antibodies / Introdu??o: Os v?rus que causam doen?as em humanos podem causar epidemias e pandemias levando ? morte de milhares de pessoas. Quando um v?rus infecta o organismo, o sistema imunol?gico ? ativado para tentar controlar essa infec??o. A principal resposta contra v?rus em geral consiste em c?lulas T CD8+ citot?xicas e os anticorpos produzidos por c?lulas B. Contudo, tanto as c?lulas B como as T CD8+ necessitam de c?lulas auxiliares T CD4+ para uma resposta eficiente, sendo que a produ??o de anticorpos precisa de um tipo especifico de c?lulas auxiliares, as c?lulas T CD4+ foliculares (TFH). Ambas as respostas podem gerar mem?ria, assim em uma segunda infec??o pela mesma cepa viral podemos responder de maneira mais eficaz. Nesses dois artigos seguintes discutiremos a fun??o de c?lulas TFH na resposta viral. Primeiramente temos um artigo que revisa todos os estudos que conseguiram fazer uma liga??o entre essa popula??o celular e v?rus diversos, como HIV, HBV, EBV, LCMV e Influenza. Ap?s temos um artigo original tentando desvendar o mecanismo pelo qual o V?rus Sincicial Respirat?rio (RSV) inibe uma resposta imunol?gica humoral adequada. Anticorpos gerados em infec??es pelo RSV frequentemente possuem baixa afinidade e pouco poder de neutraliza??o, gerando ac?mulo de imunocomplexos inflamat?rios nas vias a?reas inferiores em crian?as at? os 2 anos de idade. Isso pode levar ? morte por bronqueolite e pneumonia em cerca de 50% dos casos, e aumenta a chance de desenvolvimento de asma na vida adulta. As c?lulas TFH tem um importante papel na ajuda ?s c?lulas B, para a produ??o de anticorpos de alta afinidade, e troca de classe de imunoglobulinas. Objetivo: Entender os mecanismos imunol?gicos relacionados ?s c?lulas TFH nas respostas virais e descobrir como o RSV diminui a efici?ncia dos anticorpos. M?todos e Resultados: No primeiro artigo foi realizada uma revis?o nas bases de dados Medline/Pubmed por artigos originais publicados no per?odo de 2008 a 2014, utilizando os termos ?TFH? e ?Virus? juntos. J? no segundo artigo foi realizada uma pesquisa original com modelo animal, focada em determinar a intera??o do RSV com as c?lulas TFH. O RSV foi capaz de aumentar a quantidade de c?lulas TFH in vitro, aumentando tamb?m sua prolifera??o (Ki67) e morte (Anexina-V), diminuindo o tamanho dos centros germinativos in vivo e diminuindo a quantidade de anticorpos IgG contra ovalbumina. In vitro, o RSV induziu PD-L1 em c?lulas B e em c?lulas dendr?ticas. Conclus?o: As c?lulas TFH s?o de fundamental import?ncia para a resposta humoral viral. Diferentes tipos de v?rus t?m formas peculiares de interagir com o sistema imune e assim com as c?lulas TFH, como podemos notar no artigo de revis?o. Os resultados do artigo original sugerem que o RSV pode comprometer a fun??o de c?lulas TFH ao induzir PD-L1 em c?lulas dendr?ticas e c?lulas B, impedindo o desenvolvimento do centro germinativo e diminuindo a quantidade de anticorpos.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-05-14T20:09:46Z No. of bitstreams: 1 468627 - Texto Completo.pdf: 868809 bytes, checksum: da322f4444f2d9e11e6d453c3af66303 (MD5) / Made available in DSpace on 2015-05-14T20:09:46Z (GMT). No. of bitstreams: 1 468627 - Texto Completo.pdf: 868809 bytes, checksum: da322f4444f2d9e11e6d453c3af66303 (MD5) Previous issue date: 2015-03-12 / Tooth movement in response to orthodontic forces is results from remodeling changes that affect the periodontal ligament and the bone surrounding teeth. This movement may be faster or slower depending on the physical characteristics of the force applied and the biological response induced, and is characterized by zones of compression and tension on the periodontal ligament. Knowledge about the tissue changes generated in this process is essential to plan the movement of teeth and their adjoining structures into a new position without any damage to tissues. In this study, the literature was reviewed to improve our understanding of the cellular, biochemical and molecular phenomena that affect the structures of the periodontal ligament (PL), as well as the alveolar bone surrounding the tooth and involved in the tooth movement that is induced in the process. The purpose of our systematic review was to evaluate studies about cytokines in gingival fluid (GF) during orthodontic treatment, summarize the patterns of cytokine regulation that have been more frequently studied and discuss their clinical implications. Additionally, we investigated changes in immunological and bone modulators in response to the application of orthodontic forces. For that purpose, GF of teeth that were involved in maxillary expansion was evaluated qualitatively to measure interleukin concentrations and their predictive value as a market of inflammatory response intensity and bone turnover during orthodontic movement. The purpose of this investigation was to evaluate whether the force applied on the PL during maxillary expansion affected the composition of the gingival crevicular fluid (GCF), specifically the level of IL-17 on the sides where tension and compression were applied. GCF samples were collected from the mesiobuccal and mesiolingual areas of 32 molars of 16 patients aged 7 to 14 years who had to undergo maxillary expansion. These patients started orthodontic treatment sometime between 2012 and 2013 at the School of Dentistry of the Pontif?cia Universidade Cat?lica do Rio Grande do Sul (PUCRS), Porto Alegre, Brazil. Metal bands were placed around the permanent maxillary first molars to hold a modified Hyrax expander. One of the investigators collected clinical data about the sites listed above during activation of the expander. An enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin levels. The sites were examined to measure IL-17 before the device was placed and at 1, 7 and 14 days of active use of the expander. The forces applied resulted in an increase in IL-17 expression in periodontal tissues one day after the application of tension and compression at both measurement sites. IL-17 levels tended to be higher (p ? 0.05) seven days after the Hyrax screw was activated. From that time to the fourteenth day, there was a tendency to a decrease in IL-17 levels (p ? 0.05). We concluded that there was an increase in IL-17 expression at both tension and compression sites in response to force application. / O movimento dent?rio promovido por for?as ortod?nticas ? baseado em altera??es remodeladoras que ocorrem no ligamento periodontal e no osso que circundam os dentes. Este movimento pode ocorrer de forma mais r?pida ou lenta, dependendo das caracter?sticas f?sicas da for?a aplicada e da resposta biol?gica induzida e ? caracterizado por cria??o de zonas de compress?o e tens?o no ligamento periodontal. O conhecimento das altera??es teciduais geradas ? essencial para o planejamento do movimento do dente e de suas estruturas anexas para uma nova posi??o, sem danos aos tecidos. Neste estudo, uma revis?o de literatura teve como objetivo possibilitar a compreens?o dos fen?menos celulares, bioqu?micos e moleculares que ocorrem nas estruturas do ligamento periodontal (PDL) e, tamb?m do osso alveolar ao redor do dente e que est?o envolvidos na movimenta??o dent?ria induzida. A revis?o sistem?tica teve como objetivo avaliar estudos sobre citocinas no fluido gengival (GCF) durante o tratamento ortod?ntico, resumindo os padr?es de regula??o das citocinas mais estudadas e explorar suas implica??es cl?nicas. Al?m disso, foram investigadas altera??es dos moduladores imunol?gicos e ?sseos em resposta a aplica??o da for?a ortod?ntica. Para tanto o fluido gengival de dentes submetidos ao procedimento de expans?o maxilar foi avaliado qualitativamente verificando concentra??es de interleucinas e seu valor preditivo como marcador da intensidade da resposta inflamat?ria e turnover ?sseo na movimenta??o ortod?ntica. O objetivo da investiga??o foi avaliar se a for?a aplicada sobre o ligamento periodontal durante a expans?o maxilar se reflete na composi??o do Fluido do Sulco Gengival (FSG), mais especificamente no n?vel de interleucina IL-17 dos lados de tens?o ou compress?o. As amostras do FSG foram coletadas das regi?es mesiovestibulares e m?siopalatinas de 32 molares de 16 pacientes com idade entre 7 a 14 anos, que tinham necessidade de expans?o maxilar. Estes pacientes iniciaram o tratamento ortod?ntico durante o per?odo compreendido entre 2012 e 2013, na Faculdade de Odontologia da Pontif?cia Universidade Cat?lica do Rio Grande do Sul (PUCRS). Eles receberam an?is nos primeiro molares permanentes superiores para a confec??o de um aparelho disjuntor de Hyrax modificado. Os dados cl?nicos foram coletados por um investigador nas regi?es supracitadas durante o per?odo de ativa??o do aparelho. Empregando-se o ensaio imunoenzim?tico (ELISA) foram detectados os n?veis da interleucina. Os locais foram analisados para IL-17 antes da coloca??o do aparelho, 1, 7 e 14 dias de uso ativo do aparelho. As for?as empregadas resultaram em um aumento da express?o de IL-17 nos tecidos periodontais ap?s um dia de aplica??o em ambos os lados, de tens?o e compress?o. Os n?veis de IL-17 tenderam a ser maiores (p ? 0,05) ap?s sete dias de ativa??o do parafuso. Deste momento, at? o d?cimo quarto dia, observou-se uma tend?ncia para diminui??o dos n?veis de IL-17 (p ? 0,05). Concluiu-se que houve um aumento na express?o de IL-17 em ambos os locais, de tens?o e compress?o, em resposta ? aplica??o de for?a.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-07-01T12:16:16Z No. of bitstreams: 1 471496 - Texto Completo.pdf: 3473251 bytes, checksum: 4c713013d9c0095e2989171bafa30fb2 (MD5) / Made available in DSpace on 2015-07-01T12:16:16Z (GMT). No. of bitstreams: 1 471496 - Texto Completo.pdf: 3473251 bytes, checksum: 4c713013d9c0095e2989171bafa30fb2 (MD5) Previous issue date: 2015-03-02 / The elderly population is now the fastest growing in the world. The immune system becomes compromised with age, being less effective in fighting malignancies. An amended fee of CD8 T cells, increased compared to CD4 cells (usually more frequent in the blood) defines the concept of immune risk profile in the elderly (less than 1 ratio). In this research, we studied the immunological profile of 58 women with breast cancer, determining the ratio of CD4+/CD8+ in peripheral blood. We compare this link into groups according to age of the patient (less or more than 60 years), lymph node involvement, tumor size, the immunohistochemical profile, and the occurrence of adverse events (local recurrence, axillary recurrence and metastasis). The ratio of CD4+ /CD8+ lymphocytes average was 1.87 (1.1 to 3.32), therefore, no patient had a concept immune risk profile. Comparing the groups according to age, the presence of lymph node metastases, tumors smaller or higher than 2 cm, were no statistically significant differences. However, the CD4 / CD8 ratio in patients positive for human epidermal growth factor (HER-2 / neu) was higher than in samples of patients with hormone receptor positive carcinoma and Her-2 / neu negative (p = 0.036 ). As far as was possible to search, these are the first data on CD4+ and CD8 T+ cells specifically in a population with breast cancer. The increase in the number of CD4+/CD8+ current evidenced in these women with Her-2 carcinoma / neu positive, seems to signal an activation of the immune system in women with poor prognosis of tumors. / A popula??o de idosos ? hoje a que mais cresce no mundo. O sistema imune torna-se comprometido com a idade, sendo menos eficaz no combate a neoplasias malignas. Uma taxa alterada de c?lulas T CD8 positivo (CD8+), aumentada em rela??o ?s c?lulas CD4 positivo (CD4+) (normalmente mais freq?entes no sangue) define o conceito de perfil de risco imunol?gico no idoso (rela??o menor que 1). Nesta pesquisa, estudamos o perfil imunol?gico de 58 mulheres com c?ncer de mama, determinando a rela??o das c?lulas T CD4+/CD8+ no sangue perif?rico. Comparamos esta rela??o em grupos conforme a idade da paciente (menos ou mais que 60 anos), o comprometimento linfonodal, o tamanho do tumor, o perfil imunohistoqu?mico, e a ocorr?ncia de eventos adversos (recidiva local, axilar e met?stases). A propor??o de linf?citos T CD4+/CD8+ m?dia foi 1,87 (1,1 a 3,32), portanto, nenhuma paciente apresentou por conceito perfil imunol?gico de risco. Comparando os grupos conforme a idade, a presen?a de met?stases linfonodal, tumores menores verso maiores que 2cm n?o houveram diferen?as estatisticamente significativas. No entanto, a rela??o CD4+/CD8+ em pacientes positivos para o fator de crescimento epid?rmico humano (Her-2/neu) foi mais elevada do que em amostras de pacientes com carcinomas receptor hormonal positivos e Her-2/neu negativo (p=0,036). Tanto quanto foi poss?vel pesquisar, estes s?o os primeiros dados sobre c?lulas T CD4+ e CD8+ especificamente em uma popula??o com c?ncer de mama. O aumento do n?mero de linf?citos T CD4+/CD8+ circulante evidenciado nestas mulheres com carcinoma Her-2/neu positivo, parece sinalizar uma ativa??o do sistema imunol?gico em mulheres com tumores de pior progn?stico.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-08-13T20:17:59Z No. of bitstreams: 1 473456 - Texto Parcial.pdf: 768622 bytes, checksum: 552f69a1d4c4b1b7e4906070b478b964 (MD5) / Made available in DSpace on 2015-08-13T20:17:59Z (GMT). No. of bitstreams: 1 473456 - Texto Parcial.pdf: 768622 bytes, checksum: 552f69a1d4c4b1b7e4906070b478b964 (MD5) Previous issue date: 2015-03-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction: Asthma is a disease that affects most children, characterized by inflammation and airway hyperresponsiveness. Asthma has two main phenotypes depending on the presence of atopy. In addition to Th2 cells, Th17 cells, Th1 and Treg cells are involved in disease pathology by secreting cytokines. CD4 T cells have plasticity, which are influenced by environmental and genetic factors. Aim: To assess the phenotypic profile of CD4 T cells in asthmatic children of school age, with or without the presence of atopy and healthy controls and correlate the data with disease severity. In addition, analyze cells expressing more than one transcription factor in the blood of atopic asthmatic children to D.pteronyssinus stimulating mononuclear cells with DerP1 protein. Methods: This was a cross-sectional case-control study, where 8-14 years-old asthmatic and healthy children were recruited of public schools in Porto Alegre/RS. Mononuclear cells were isolated by Histopaque peripheral blood of children and cultured with anti-CD3/CD28 or DerP1 protein. After 24 hours the cells were stained with antibodies specific for transcription factors for analysis of Th2, Th17, Th1 and Treg cells by flow cytometry. Moreover, it was analyzed in plasma allergen-specific IgE specific. Results: The results showed that children with asthma have a higher frequency of CD4+ GATA-3+ compared to controls. Frequency of CD4+GATA-3+ROR?t+ cells correlated with disease severity, except with severe therapy resistant asthma (STRA). Atopic patients showed a higher proportion of co-expressing more than one transcription factor cells compared to non-atopic individuals. Stimulation with DerP1 protein enhances the frequency of CD4 + Foxp3 +ROR?t+ compared with the control and stimulation with anti-CD3/ CD28. Conclusion: In general, the results presented in this thesis allowed us to conclude that the allergens play an important role in the development of cells expressing more than one transcription factor in atopic patients. In addition, our study was the first to demonstrate that STRA children have a distinct expression profile of regulatory transcription factors of CD4 T cells Th1, Th2, Th17 and Treg compared to other severity levels of the disease. / Introdu??o: A asma ? uma doen?a prevalente em crian?as, tendo como principal caracter?stica a inflama??o e hiperresponsividade das vias a?reas. Al?m das c?lulas Th2, as c?lulas Th17, Th1 e Treg s?o envolvidas na patologia da doen?a pela fun??o que elas e as citocinas por elas secretadas exercem. A asma apresenta dois fen?tipos principais, os quais dependem da presen?a de atopia. As c?lulas TCD4 apresentam plasticidade, que s?o influenciadas por fatores ambientais e gen?ticos. Objetivos: Avaliar o perfil fenot?pico de c?lulas TCD4 de crian?as em idade escolar asm?ticas, com ou sem a presen?a de atopia e de controles saud?veis. Correlacionar os dados obtidos com a severidade da doen?a. Al?m disso, avaliar as c?lulas que co-expressam mais de um fator de transcri??o no sangue de crian?as asm?ticas at?picas para D.pteronyssinus estimulando as c?lulas com a prote?na Derp1. Materiais e m?todos: Este foi um estudo transversal caso-controle, onde crian?as de 8 a 14 anos foram recrutadas de escolas p?blicas de Porto Alegre/RS, asm?ticas e controles. As c?lulas mononucleares foram isoladas por Histopaque do sangue perif?rico destas crian?as e colocadas em cultura com anti-CD3/CD28 ou com prote?na Derp1. Ap?s 24hs as c?lulas foram marcadas com anticorpos espec?ficos para os fatores de transcri??o para an?lise do perfil Th2, Th17, Th1 e Treg por citometria de fluxo. Al?m disto, foi analisado no plasma a IgE espec?fica para um painel de alergenos. Resultados: Os resultados obtidos mostraram que as comparado com os controles. As c?lulas duplo-positivas CD4+GATA-3+ROR?t+ correlacionaram com o grau de severidade da doen?a, exceto com asma severa de dif?cil controle (ADC). Pacientes at?picos apresentaram uma propor??o maior de c?lulas co-expressando mais de um fator de transcri??o comparados com indiv?duos n?o-at?picos. Estimula??o de c?lulas provenientes de pacientes at?picos com a prote?na Derp1 aumenta a frequ?ncia de c?lulas CD4+Foxp3+ROR?t+ quando comparados com o controle e estimula??o com anticorpos anti-CD3 e anti-CD28. Conclus?o: De uma maneira geral, os resultados apresentados nesta tese nos permitem concluir que os al?rgenos desempenham um papel importante no desenvolvimento das c?lulas que expressam mais de um fator de transcri??o em pacientes at?picos. Al?m disso, nosso estudo foi o primeiro a demonstrar que crian?as com ADC apresentam um perfil distinto de express?o de fatores de transcri??o reguladores de c?lulas TCD4 do tipo Th1, Th2, Th17 e Treg quando comparado com crian?as com doen?a menos severa.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-10-28T21:42:44Z No. of bitstreams: 1 475919 - Texto Parcial.pdf: 381957 bytes, checksum: c63b236b74973994f585b6bd7db08855 (MD5) / Made available in DSpace on 2015-10-28T21:42:44Z (GMT). No. of bitstreams: 1 475919 - Texto Parcial.pdf: 381957 bytes, checksum: c63b236b74973994f585b6bd7db08855 (MD5) Previous issue date: 2015-07-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction : Respiratory Syncytial Virus (RSV) is a major cause of viral lower respiratory tract infection in children under two years of age. Memory CD8 T cell response to VSR does not provide an efficient and long-lasting immune response, so there are recurrent infections throughout life. The role of mTOR (mammalian target of rapamycin) and the purinergic receptor P2X7 in the memory CD8 T cell response during RSV infection has not been investigated. Objectives : To analize the effect of rapamycin on dendritic cells (DCs) during RSV infection, as well as to evaluate the role of P2X7 purinergic receptor in CD8 memory T cells response during RSV infection. Methodology : Bone marrow derived dendritic cells (BMDCs) differentiated from C57BL/6 P2X7-/- and C57BL/6 mice were infected with VSR virus and used to activate T cells purified from C57BL/6 P2X7-/- and C57BL/6 mice in vitro for 96 hours. In addition, BMDCs differentiated from C57BL/6 mice received 20ng/ml rapamycin during 1h prior infection. The following parameters were evaluated: BMDC cell death by apoptosis, memory cells markers by flow cytometry and RNA viral quantitation was performed by real time PCR. Results : Rapamycin treatment in RSV infected BMDCs decreases the frequency of CD8+CD44high cells and increases the level of viral RNA in DCs, but the rapamycin does not affect the viability of the infected dendritic cells. Furthermore, when BMDCs were treated with rapamycin, an increase of BMDC survival occurred, depending on the contact with the T cells. The absence of purinergic receptor P2X7 in T cells leads to a decrease in the frequency of CD8+CD122+KLRG1-, however the absence of purinergic receptor in infected BMDCs increases the frequency of CD8+CD122+ KLRG1- T cells. Conclusion : Our study suggests that P2X7 receptor is involved in memory CD8 T cell response. In addition our data indicated that mTOR inhibition increases the survival of BMDCs in a mechanism dependent on T-cell contact and also suggests that rapamycin treatment on dendritic cells during VSR infection affect CD8 T cell differentiation. / Introdu??o : O V?rus Sincicial Respirat?rio (VSR) ? um dos principais causadores de infec??o viral do trato respirat?rio inferior em crian?as menores de dois anos de idade. A resposta de c?lulas T CD8 de mem?ria para VSR n?o apresenta uma resposta imune eficiente e duradoura, por isso s?o recorrentes as infec??es durante a vida. At? o momento n?o se tem o conhecimento sobre o envolvimento do mTOR (mammalian target of rapamycin) e do receptor purin?rgico P2X7 na resposta de c?lulas T CD8 de mem?ria durante a infec??o de VSR. Objetivos : Investigar o efeito da rapamicina nas c?lulas dendr?ticas (DCs) durante a infec??o viral, bem como, avaliar o efeito do receptor purin?rgico P2X7 nas c?lulas dendr?ticas (DCs) infectadas com o v?rus VSR na resposta de c?lulas T CD8 de mem?ria. Metodologia : C?lulas dendr?ticas diferenciadas de medula ?ssea (BMDCs) de camundongos C57BL/6 P2X7-/- e C57BL/6 infectadas com VSR foram utilizadas para ativar c?lulas T purificadas de camundongos C57BL/6 P2X7-/- e C57BL/6 durante 96 horas. Al?m disso, um grupo de BMDCs de camundongos C57BL/6 recebeu 20ng/mL de rapamicina durante 1h antes da infec??o. As DCs foram marcadas para investiga??o de morte celular por apoptose, as c?lulas T marcadas para an?lise c?lulas de mem?ria e a quantifica??o do RNA viral foi realizada atrav?s de PCR em tempo real. Resultados : O tratamento com o inibidor de mTOR nas BMDCs infectadas pelo VSR diminuiu a gera??o de c?lulas T CD8+ CD44high e aumentou os n?veis de RNA viral nas DCs, por?m a rapamicina n?o influenciou a viabilidade das c?lulas dendr?ticas infectadas. Quando as BMDCs foram tratadas com rapamicina, houve um aumento de sobreviv?ncia das c?lulas, dependente do contato com as c?lulas T. Na aus?ncia do receptor purin?rgico P2X7 nas c?lulas T, ocorreu uma diminui??o na frequ?ncia das c?lulas T CD8+CD122+KLRG1-, entretanto a aus?ncia do receptor purin?rgico nas BMDCs infectadas aumentou a frequ?ncia destas c?lulas T CD8+CD122+KLRG1-. Conclus?o : Nosso estudo sugere que o receptor P2X7 est? envolvido na resposta de c?lulas T CD8 de mem?ria durante a infec??o pelo VSR. Al?m disso, sugere-se que as c?lulas dendr?ticas tratadas com rapamicina durante a infec??o com VSR prejudica a diferencia??o de c?lulas T CD8.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2015-11-05T12:26:05Z No. of bitstreams: 1 476006 - Texto Completo.pdf: 2870247 bytes, checksum: 76076f40e5be705d3be21ff4d7ceb1f3 (MD5) / Made available in DSpace on 2015-11-05T12:26:06Z (GMT). No. of bitstreams: 1 476006 - Texto Completo.pdf: 2870247 bytes, checksum: 76076f40e5be705d3be21ff4d7ceb1f3 (MD5) Previous issue date: 2015-07-29 / Aging has been associated with increased generation of free radicals as well as immunosenescence. Previous studies have identified older individuals with inverted T CD4:CD8 cell ratio and increased immunity to cytomegalovirus (CMV). Here, we investigated markers of oxidative stress and antioxidant defences in older individuals with inverted CD4:CD8 T-cell ratio. From 421 healthy community-dwelling older adults recruited, 61 subjects were identified with inverted CD4:CD8 T-cell ratio. Older individuals with a CD4:CD8 ratio < 1 had increased levels of plasma advanced oxidation protein products (AOPP) and Ferric Reducing Ability of Plasma (FRAP), but reduced levels of thiobarbituric acid reactive substances (TBARS) as compared to subjects with normal CD4:CD8 T-cell ratio. The CMV IgG serology was negatively correlated with CD4:CD8 ratio. These markers were more evident among elderly men than women. Our data suggest a close relationship between chronic CMV infection and oxidative profile in older individuals in the midst of its influence on the peripheral T-cell subsets. / O envelhecimento tem sido associado com o aumento da gera??o de radicais livres, bem como com a imunossenesc?ncia. Estudos anteriores j? haviam identificado os idosos que apresentavam a invers?o da raz?o CD4:CD8 e a sorologia mais acentuada para citomegalov?rus (CMV). Aqui, investigamos marcadores de estresse oxidativo e as defesas antioxidantes em idosos com a rela??o invertida entre as c?lulas T CD4 e CD8. De um total de 421 idosos recrutados na comunidade, 61 sujeitos foram identificados com a invers?o da raz?o CD4:CD8. Os idosos com CD4:CD8<1 tinham n?veis aumentados de produtos proteicos de oxida??o avan?ada (AOPP) e atividade antioxidante total pelo m?todo de redu??o do ferro (FRAP) no plasma, mas n?veis reduzidos de subst?ncias reativas ao ?cido tiobarbit?rico (TBARS), em compara??o com indiv?duos com raz?o CD4:CD8 normais. A sorologia para CMV (IgG) foi negativamente correlacionada com a raz?o CD4:CD8. Estes marcadores foram mais evidentes entre os homens do que as mulheres idosas. Nossos dados sugerem uma estreita rela??o entre a infec??o por CMV cr?nica e perfil oxidativo em indiv?duos mais velhos no meio de sua influ?ncia sobre os subtipos de c?lulas T perif?ricas.

Made available in DSpace on 2015-04-14T13:33:08Z (GMT). No. of bitstreams: 1 451963.pdf: 1054634 bytes, checksum: 22a1a79881824872b314b26c9cf9ec20 (MD5) Previous issue date: 2013-03-04 / Introduction: Asthma is a disease with predominant presence of CD4 Th2 cells that affects children. The disease main characteristics is inflammation and airway hyperreactivity. Asthma has two main phenotypes depending on the presence of atopy. It was not yet fully described in asthmatic children of school age the presence of different profiles of Th1, Th2, Th17 and Treg cells in peripheral blood, which may be related to the pathogenesis of the disease. Aims: To analyze the phenotypic of CD4 T cells in asthmatics children in Porto Alegre/RS. Methods: This was a croos-sectional study when it was recruited children between 9 to 15 years old, asthmatics and controls. The PBMCs were isolated using Ficoll and the cells were cultured with anti-CD3/CD28antibodies, Derp1 or left unstimulated. After 24hs the cells were stained with antibodies specifics for transcription factors in order to analyze Th1, Th2, Treg and Th17 cells by flow cytometry and the supernatant was collected for cytokine analysis by CBA assay. Plasma was used to perform analysis of specific IgE. Results: It was analyzed 104 patients, 12 controls and 92 asthmatics. Atopic asthmatic patients presented higher frequency of Th2 cells compared with non-atopic and atopic controls. When we analyzed cells expressing more than one transcription factor it was observed that atopic asthmatics patients have a higher frequency of CD4+GATA3+FOXP3+, CD4+ROR&#947;T+GATA3+ and CD4+GATA3+Tbet+ compared with non-atopic patients and controls atopic. Analyzing the asthma severity it was seen a higher frequency of CD4+ROR&#947;T+GATA3+ in patients with moderate asthma compared with patients with mild asthma. Stimulating the cells from atopic asthma patients with anti-CD3 and anti-CD28 induced more Th1 profile while stimulating with DerP1 protein changed the profile of the cells mainly to Th17 and Treg cells. Conclusion: This is the first study in children that analyzes Th1, Th2, Treg and Th17 cells using the tag transcription factor by flow cytometry in asthmatic or atopic and non-atopic controls. We found a Th2 profile in atopic asthmatic children according to the literature. Interestingly non-atopic children showed no predominant profile. Children with moderate asthma have a profile that expresses both the transcription factor GATA3 and ROR&#947;T. Depending on the stimulus used it was induced different phenotypes in the cells of this group of patients with asthma. / Introdu??o: A asma ? uma doen?a com predomin?ncia da presen?a das c?lulas T CD4 do tipo Th2, que afeta das crian?as e tem como principais caracter?sticas a inflama??o e hiperreatividade das vias a?reas. A asma apresenta dois fen?tipos principais dependendo da presen?a de atopia. Ainda n?o foi totalmente descrito em crian?as asm?ticas com idade escolar a presen?a dos diferentes perfis de c?lulas Th1, Th2, Th17 e Treg no sangue perif?rico, que podem estar relacionados com a patog?nese da doen?a. Objetivo: Avaliar o perfil fenot?pico de c?lulas T CD4 em pacientes asm?ticos escolares do munic?pio de Porto Alegre/RS. M?todos: Este foi um estudo transversal realizado com o recrutamento de crian?as com 09 a 15 anos de idade, asm?ticas e controles. O sangue perif?rico foi coletado e as c?lulas mononucleares foram separadas utilizando Ficoll e colocadas em cultura com anticorpos anti-CD3/CD28. Ap?s 24hs as c?lulas foram marcadas com anticorpos espec?ficos para fatores de transcri??o para an?lise do perfil Th1, Th2, Th17 e Treg por citometria de fluxo e o sobrenadante foi recolhido para analise de citocinas por CBA. No plasma foi realizada a an?lise de IgE espec?ficas. Resultados: Foram analisados 104 pacientes, sendo 12 controles e 92 asm?ticos. Os pacientes asm?ticos at?picos apresentaram um perfil Th2 mais acentuado, comparado com os n?o-at?picos e controles at?picos. Quando analisamos as c?lulas que expressam mais de um fator de transcri??o foi observado que os pacientes asm?ticos at?picos apresentam maior frequ?ncia de c?lulas T CD4 positivas GATA3+FOXP3+, ROR&#947;T+GATA3+, GATA3+Tbet+ comparada com pacientes n?o-at?picos e controles at?picos. Analisando a severidade foi observado uma frequencia maior de c?lulas T CD4+ ROR&#947;T+ GATA3+ em pacientes com asma moderada comparado com pacientes com asma leve. O est?mulo com anti-CD3 e anti-CD28 nas c?lulas dos pacientes asm?ticos at?picos induziu mais um perfil Th1 enquanto que est?mulo com DerP1 mudou o perfil de algumas c?lulas principalmente de c?lulas Tregs e Th17. Conclus?o: Este ? o primeiro estudo em crian?as que analisa o perfil Th1, Th2, Th17 e Treg utilizando a marca??o de fatores de transcri??o por citometria de fluxo em crian?as asm?ticas at?picas ou n?o-at?picas e controles. Encontramos um perfil mais Th2 em crian?as asm?ticas at?picas de acordo com a literatura. Interessantemente as crian?as n?o-at?picas n?o apresentaram nenhum perfil predominante. As crian?as com asma moderadas apresentam um perfil que expressa ao mesmo tempo o fator de transcri??o RORgT e GATA3. Dependendo do estimulo s?o induzidas diferentes fen?tipos nas c?lulas deste grupo de pacientes com asma.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-06-17T19:39:46Z No. of bitstreams: 1 TES_ALINE_DEFAVERI_DO_PRADO_COMPLETO.pdf: 1737423 bytes, checksum: 77ee581083f58c46dd10a545b62097a5 (MD5) / Made available in DSpace on 2016-06-17T19:39:46Z (GMT). No. of bitstreams: 1 TES_ALINE_DEFAVERI_DO_PRADO_COMPLETO.pdf: 1737423 bytes, checksum: 77ee581083f58c46dd10a545b62097a5 (MD5) Previous issue date: 2016-03-28 / Introduction: Rheumatoid Arthritis (RA) is an autoimmune, inflammatory and chronic disease. Muskuloskeletal ultrasound (MSUS) has been increasingly used for diagnostic evaluation and monitoring of patients. Regulatory T cells (Tregs) and lymphocytes producers of IL 17 (Th17) imbalance and disfuntion, as well as pro inflammatory cytokines, have been implicated in the pathogenesis of RA. There are few studies on the association of circulating lymphocites subtypes and cytokines with MSUS findings in RA. Methods: One hundred and one RA patients (1987 American College of Rheumatology criteria) treated with disease-modifying antirheumatic drugs (DMARDs) were included in this cross sectional study. A blood sample was taken just before clinical and ultrasonographic evaluation, which were all performed on the same day, consecutively and in a blinded fashion. Lymphocytes were isolated and immunophenotyped by flow cytometry to investigate regulatory FoxP3+ T cells and IL-17+ cells. Plasma Th1-Th2-Th17 cytokines (IL-2, IL-4, IL-6, IL-10, IL-17, TNF e IFN-?) and VEGF were searched using a Cytometric Bead Array (CBA; BD biosciences) kit by flow cytometry. Disease acitivity and disability were measured using Disease Activity Score in 28 joints (DAS28) and Health Assessment Questionnaire (HAQ). MSUS (MyLab 60, Esaote, Genova, Italy, 18 MHz linear probe) was performed consecutively by two ultrassound-trained rheumatologists on the wrists, 2th and 3th metacarpophalangeal and 2th and 3th proximal interphalangeal joints of both hands. Gray-scale synovial hypertrophy (PS) and power Doppler signal (pD) were searched using a semi-quantitative scale (0-3). Erosions were classified as present or absent. The sum of the individual joint scores for PS and pD (10-joint PS and pD scores) was calculated and used to correlate with clinical and laboratory data. Mann-Whitney test, Kruskal-Wallis test and Spearman correlation coefficient (rS) were used for statistical analysis, as well as liner multivariate regression. Interater agreement was tested using kappa statistics and intraclass correlation. Results: Among 101 patients, we were able to measure Treg/Th-17 in 90 individuals. Plasma cytokines were searched in 64 patients. Clinical and demographic features were: mean age, 55.8 ? 11 years; female gender, 80%; Caucasians, 85%; median (interquartile range) disease duration 6 (2-13) years; mean ? SD DAS28, 4.28 ? 1.64; mean ? SD HAQ score, 1.11 ? 0.85. Interobserver agreement (kappa) for US features varied from 0.53 to 1.0. Intraclass correlation for 10-joint PS score was 0.964 (95% CI 0.899-0.986, P <0.000) and for 10-joint pD score was 0,859 (95% CI 0.646-0.941, P <0.001). There was no significant correlation of 10-joint PS and pD scores with DAS28 and HAQ score. The presence of bone erosions was associated with 10-joint PS and pD scores (p=0.002), but not with DAS 28 (p=0.079) or HAQ (0.057). Swollen joint count, but not tender joint count, was correlated with 10-joint PS and pD scores (rS=0.54, P<0.001 and rS=0.39, P<0.001; respectively), as well as associated with bone erosions (P<0.001). There was no significant correlation of with 10-joint PS and pD scores with peripheral Tregs (rS=0.122, P=0.254 and rS=0,056, P=0.602) and Th17 cells (rS=-0.083, P=0.438 and rS=-0.060, P=0,575). Tregs and Th17 cells were not associated with erosions (p= 0,831 and p=0,632, respectively). Among all tested cytokines, IL-6 was correlated with DAS28 (rs 0.31 IC95% 0.07 to 0.52), eritrocyte sedimentation rate (rs 0.43 IC95% 0.19 to 0.62) and swollen joint count (rs 0.39 95%CI 0.15 a 0.59). IL-6 was also correlated with 10-joint pD score (rs 0.33 IC95% 0.07 to 0.56), right and left wrists pD (rs 0.34 IC95% 0.11 to 0.54 and rs 0.45 IC95% 0.21 to 0.64), and right and left PS (rs 0.40 IC95% 0.20 to 0.59 and rs 0.35 IC95% 0.08 to 0.57). Using multivariate linear regression model, 10-joint pD score was positively associated with IL-6 independently of DAS28 (P=0.025). There was no association of any of the tested cytokines with bone erosions (P? 0.17 for all tests). Conclusions: In established RA patients, treated with non biological DMARDs, we observed the following: lack of correlation of 10-joint PS and pD scores and DAS28 and HAQ; positive association of 10-joint PS and pD scores with bone erosions; positive association of swollen joint count, but not tender joint count, with MSUS synovitis and erosions; lack of correlation of MSUS features and circulating Treg and Th-17 cells; positive correlation of plasma IL-6 and MSUS synovitis. The association of IL-6 with 10-joint pD score was independent of DAS28. / Introdu??o. A artrite reumatoide (AR) ? uma doen?a autoimune inflamat?ria cr?nica com envolvimento articular proeminente. O ultrassom de alta resolu??o (US) tem sido empregado de maneira crescente na avalia??o diagn?stica e monitoriza??o da doen?a. O desequil?brio entre as fun??es de linf?citos T regulat?rios (Treg) e Th-17, bem como o papel de citocinas pr? inflamat?rias s?o centrais em sua patog?nese. H? escassez de estudos sobre a associa??o entre os achados ultrassonogr?ficos de atividade da AR e dano estrutural com subtipos linfocit?rios e citocinas no sangue perif?rico. M?todos: Neste estudo transversal, foram inclu?dos 101 pacientes com AR de acordo com os crit?rios de 1987 em tratamento com drogas remissivas n?o-biol?gicas. Uma amostra de sangue foi coletada imediatamente antes das avalia??es cl?nica e ultrassonogr?fica, que foram feitas todas no mesmo dia, em sequ?ncia. Os linf?citos foram isolados e imunofenotipados por citometria de fluxo para identificar c?lulas T reg FoxP3+ e linf?citos produtores de IL-17. Citocinas do perfil Th1-Th2-Th17 (IL-2, IL-4, IL-6, IL-10, IL-17, TNF e IFN-?) e VEGF foram quantificadas no plasma por citometria de fluxo utilizando-se o kit Cytometric Bead Array (CBA; BD biosciences). Atividade da doen?a foi quantificada por DAS28 e capacidade funcional, pelo HAQ, ambos realizados por reumatologista treinado, cegado em rela??o aos achados de US. Dois reumatologistas (cegados em rela??o aos dados cl?nicos) com treinamento em ultrassonografia realizaram avalia??o de sinovite e presen?a de eros?es em punhos, 2? e 3? articula??es metacarpofalangeanas e 2? e 3? interfalangeanas proximais bilateralmente utilizando aparelho de alta resolu??o (MyLab 60, Esaote, It?lia, transdutor linear de 18 mHz). Prolifera??o sinovial na escala de cinzas (PS) e capta??o de power Doppler (pD) foram avaliadas utilizando uma escala semi-quantitativa com varia??o de 0 a 3. Eros?es foram classificadas como presentes ou ausentes. A soma dos escores individuais de PS e pD foi calculada (escore 10 PS e escore 10 pD) e utilizada para correlacionar com dados cl?nicos e de laborat?rio. Teste de Mann-Whitney, Kruskal-Wallis e coeficiente de correla??o de Spearman foram usados na an?lise estat?stica, bem como regress?o linear m?ltipla. Correla??o intraclasse e estat?stica kappa foram usados para concord?ncia interobservador. Resultados: Dentre os 101 pacientes inclu?dos, obtivemos quantifica??o de c?lulas Treg e Th-17 em 90 pacientes e dosagem de citocinas em 64 indiv?duos. Em sua maioria, a amostra foi composta por mulheres (80%) da ra?a branca (85%), com m?dia de idade de 55,8 anos (?11.1anos), tempo de dura??o de doen?a de 6 (2-13) anos (mediana e IIQ). A m?dia do DAS 28 (VSG) foi de 4.28 (?1.64) e do HAQ 1.11 (?0.85). Valores de kappa para a concord?ncia interobservador para o exame ultrassonogr?fico variaram de 0.53 a 1; a correla??o intraclasse para o escore 10 PS foi de 0.964 (IC95% 0.899-0.986, P <0.001) e, para o escore 10 pD, 0.859 (IC95% 0.646-0.941, P <0.001). N?o houve correla??o entre escore 10 PS e escore 10 pD com DAS28 e HAQ. Eros?es ?sseas foram associadas ao escores 10 PS e pD (p=0.002), mas n?o com DAS 28 (P=0,079) e HAQ (P =0,057). Observamos correla??o entre contagem de articula??es edemaciadas com escore 10 PS e escore 10 pD (rS=0.54, P<0.001 e rS=0.39, P<0.001; respectivamente), bem como associa??o com a presen?a de eros?es (P<0.001). N?o observamos correla??o entre contagem de articula??es dolorosas com escore 10 GS (rS = -0.071 P = 0.524), escore 10 pD (rS = -0.196 P = 0.078) ou associa??o com presen?a de eros?es (P=0.248). N?o houve correla??o significativa de escore 10 PS e pD com Treg (rS=0.122, P=0.254 e rS=0,056, P=0.602, respectivamente) e Th-17 perif?ricas (rS=-0.083, P=0.438 e rS=-0.060, P=0,575, respectivamente). N?o se observou associa??o entre percentual de c?lulas Treg e Th-17 e presen?a de eros?es (P=0.831 e P=0.632, respectivamente). A concentra??o de IL-6, mas n?o de outras citocinas, se correlacionou ao DAS28 (rs 0.31 IC95% 0.07 a 0.52), VSG (rs 0.43 IC95% 0.19 a 0.62) e contagem de articula??es edemaciadas (rs 0.39 95%CI 0.15 a 0.59), bem como com escore 10 pD (rs 0.33 IC95% 0.07 a 0.56), capta??o de pD em punhos direito e esquerdo (rs 0.34 IC95% 0.11 a 0.54 e rs 0.45 IC95% 0.21 a 0.64, respectivamente) e PS em punhos direito e esquerdo (rs 0.40 IC95% 0.20 a 0.59 e rs 0.35 IC95% 0.08 a 0.57, respectivamente). Utilizando-se modelo de regress?o linear m?ltipla, observou-se que o escore 10 pD foi positivamente associado a IL-6 independente de DAS28 (P=0.025). N?o houve associa??o entre nenhuma das citocinas testadas e a presen?a de eros?es ?sseas (P? 0.17 para todos os testes). Conclus?es: Em pacientes com AR estabelecida sob uso de remissivos sint?ticos, observamos: aus?ncia de associa??o de escore 10 PS e pD com DAS28 e HAQ; associa??o entre escore 10 PS e pD e eros?es ?sseas; associa??o de contagem de articula??es edemaciadas, mas n?o de articula??es dolorosas, com sinovite e eros?es no US; aus?ncia de associa??o entre achados de US e linf?citos Treg e Th-17 no sangue perif?rico; associa??o entre IL-6 plasm?tica e achados de sinovite no US. A associa??o entre IL-6 e escore pD ocorreu independente do DAS28.

Made available in DSpace on 2015-04-14T14:51:27Z (GMT). No. of bitstreams: 1 450693.pdf: 903446 bytes, checksum: 6805922c966dfb108b00594643df2744 (MD5) Previous issue date: 2013-03-27 / The elderly population, characterized as people over 60 years old, has grown rapidly in recent years in Brazil, and with this increase in life expectancy, age-related changes appear, for example, immunosenescence. Some studies have shown that there are differences in the immunological profile of the elderly, often related to the numbers of T cells. The immune risk profile (IRP) has been defined as CD4/CD8 T-cell ratio <1 and positivity for serum anti-cytomegalovirus (CMV) antibodies. In this study, we investigated patterns of activation and differentiation of T lymphocytes by flow cytometry in an elderly population in IRP, compared with elderly people without risk profile (non-IRP), seeking to identify additional markers for this risk profile immune. For this study we recruited 60 elderly SUS patients in Porto Alegre, aged 60 years, 30 IRP and 30 non-IRP. Lymphocytes were isolated and stimulated in vitro to evaluate the production of cytokines and the expression of transcription factors. The subpopulations of T lymphocytes found were consistent with previous studies in which IRP patients had fewer naive and central memory T cells, as well as an increase was observed in the percentage of T cells CD8 + CD28-T cells and CD8 + PD-1 + in the IRP individuals. Furthermore, there was a reduction in mean fluorescence intensity (MFI) of transcription factors to canonical subtypes of T cell help (CD4 +) in IRP patients when compared to patients without IRP. The percentages of the different subtypes (TH1, TH2, TH17; Treg) of these cells were not different between the two groups. After stimulation with anti-CD3/anti-CD28 antibodies in culture for 24 hours, the concentration of cytokines in culture supernatants increased, but no significant differences between the two groups were found, except for IL-10, which was higher in IRP patients before stimulation. These results suggested that transcriptional mechanisms important in the differentiation of helper T cells are altered in IRP patients, which may affect the immune response of these individuals. / A popula??o de idosos, caracterizada como pessoas com mais de 60 anos, vem aumentando muito nos ?ltimos anos no Brasil, e com esse aumento da expectativa de vida surgem altera??es relacionadas com a idade, por exemplo, a imunossenesc?ncia. Alguns estudos t?m demonstrado que existem diferen?as no perfil imunol?gico dos idosos, frequentemente relacionadas ao n?mero de c?lulas T. O perfil de risco imune (IRP) tem sido definido como taxa de c?lulas T CD4/CD8<1 e soro positividade para citomegalov?rus (CMV). Neste estudo, n?s investigamos padr?es de ativa??o e diferencia??o de linf?citos T, por citometria de fluxo, em uma popula??o idosa IRP, comparando com a popula??o idosa sem perfil de risco (n?o-IRP), buscando identificar marcadores adicionais para esse perfil de risco imune. Foram recrutados para esse estudo 60 idosos da rede SUS (sistema ?nico de sa?de) de Porto Alegre, com idade superior a 60 anos, sendo 30 IRP e 30 n?o-IRP. Linf?citos foram isolados e estimulados in vitro para avaliar a produ??o de citocinas e a express?o de fatores de transcri??o. As subpopula??es de linf?citos T encontradas foram condizentes com estudos anteriores em que os pacientes IRP possu?am menos c?lulas T naive e de mem?ria central, assim como foi observado um aumento nas porcentagens das c?lulas T CD8+CD28- e das c?lulas T CD8+PD-1+. Al?m disso, foi observada uma redu??o na m?dia de intensidade de fluoresc?ncia (MFI) dos fatores de transcri??o can?nicos para subtipos de c?lulas T de ajuda (CD4+) em pacientes IRP quando comparados com pacientes n?o-IRP. As porcentagens dos diferentes subtipos (TH1; TH2; TH17; Treg) dessas c?lulas n?o foram diferentes entre os dois grupos. Ap?s estimula??o com anticorpos anti-CD3/anti-CD28 em cultura por 24h, a concentra??o de citocinas no sobrenadante das culturas aumentou, mas n?o houve diferen?a significativa entre os dois grupos em rela??o ? concentra??o das citocinas em geral, exceto na IL-10 antes da estimula??o, que foi maior em pacientes IRP. Esses resultados sugerem que mecanismos transcricionais importantes para a diferencia??o de c?lulas T de ajuda est?o alterados em pacientes IRP, o que pode afetar a resposta imune desses indiv?duos.

Submitted by PPG Gerontologia Biom?dica (geronbio@pucrs.br) on 2018-09-26T11:04:34Z No. of bitstreams: 1 Trintinaglia_Lauren_Dis.pdf: 3983976 bytes, checksum: 466d953c5edcd91085fcab766ad7abd9 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-09-27T12:52:20Z (GMT) No. of bitstreams: 1 Trintinaglia_Lauren_Dis.pdf: 3983976 bytes, checksum: 466d953c5edcd91085fcab766ad7abd9 (MD5) / Made available in DSpace on 2018-09-27T12:59:22Z (GMT). No. of bitstreams: 1 Trintinaglia_Lauren_Dis.pdf: 3983976 bytes, checksum: 466d953c5edcd91085fcab766ad7abd9 (MD5) Previous issue date: 2018-03-07 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction: Individuals who have experienced childhood maltreatment (CM) present a higher sensitive profile to stress. This profile have been associated with dysregulation of the immune system, characterized by reactivation of herpesvirus and increased cellular senescence markers. New traumatic or stressful experiences, such as breast cancer diagnosis, in those sensitized individuals can trigger an increase in stress levels possibly leading to important immunological changes. Here we investigate the presence of immunosenescence markers in women newly diagnosed with breast cancer with and without history of CM. Methods: Twenty-nine women newly diagnosed with breast cancer, without start the treatment, were recruited. Fifteen with history of CM (CM+) and fourteen without history of CM (CM-). Twenty-seven women without breast cancer and CM were selected as the control group. Peripheral blood was assessed by lymphocyte subsets by flow cytometry (B cells, CD4+, CD8+, NK cells, activated T cell, regulatory T cell, early and intermediated T cell, senescent and exhaustion T cells. CMV serology was determinate by ELISA method. Results: The groups CM+ and CM- present similar cancer stage and family history of breast cancer. In peripheral lymphocyte subpopulations we found significantly reduced Early-differentiated T cell (p <0.0001), while intermediate-differentiated T cell (p<0.0001), senescence T cell (p<0.0001) and exhaustion t cells (p<0.0001) were increased in CM+ and CM? patients. The mean fluorescent intensity (MFI) was analyzed and the CD27 expression on CD4 T cells was found lower in controls as compared to CM+ and CM? groups (p = 0.002).There was no difference of CMV IgG levels between CM + and CM? groups (p=0.24), but between controls and CM groups, this association becomes significant (p= 0.008) . Conclusion: Our findings suggest that the presence of CM is not directly related with immunosenescence in women newly diagnosed with breast cancer. However, when evaluated women with breast cancer and healthy controls, this senescent profile is evident. Future longitudinal studies are necessary to explore the role of senescent cells in the disease progression and treatment response. / Introdu??o: Indiv?duos que foram expostos a maus tratos na inf?ncia apresentam um perfil de sensibilidade ao estresse mais acentuado, quando comparado a aqueles que cresceram em um ambiente adequado. Esse perfil est? relacionado com a desregula??o do sistema imunol?gico, sendo caracterizado pela presen?a de um aumento de marcadores de senesc?ncia celular e reativa??es de infec??es virais latentes. O acontecimento de novas experi?ncias traum?ticas, como o diagn?stico do c?ncer de mama, pode acarretar em um aumento dos n?veis relacionados ao estresse, provocando altera??es imunol?gicas importantes. Nesse estudo n?s investigamos a presen?a de marcadores de imunossenesc?ncia em mulheres rec?m diagnosticadas com c?ncer de mama com ou sem hist?rico de c?ncer de mama. M?todos: Vinte e nove pacientes rec?m diagnosticadas com c?ncer de mama foram recrutadas para esse estudo, sendo que quinze mulheres reportaram presen?a de maus tratos na inf?ncia (CM+), enquanto quatorze n?o vivenciaram situa??es envolvendo abuso ou neglig?ncia nesse mesmo per?odo (CM-). Para compor o grupo controle, foram selecionadas vinte e sete mulheres sem c?ncer de mama e que negaram a presen?a de maus tratos. Os linf?citos foram isolados das c?lulas mononucleares do sangue perif?rico (PBMCs), e imunofenotipados por citometria de fluxo para investigar a presen?a dos seguintes subgrupos: c?lulas B, CD4+, CD8+, c?lulas NK, c?lulas T ativadas, c?lulas T reguladoras, c?lulas T rec?m diferenciadas, c?lulas T intermedi?rias, c?lulas T senescentes e com perfil de exaust?o. A sorologia IgG para citomegalov?rus (CMV) foi realizada atrav?s do m?todo ELISA. Resultados: Os grupos CM+ e CM- n?o apresentaram diferen?as significativas em rela??o ao estadiamento da doen?a ou hist?rico familiar. Nas subpopula??es de linf?citos encontramos uma redu??o significativa de c?lulas T rec?m diferenciadas (p <0.0001), nos grupos CM+ e CM-, enquanto c?lulas T intermedi?rias (p <0.0001), senescentes (p <0.0001) e de exaust?o (p <0.0001) apresentaram-se aumentadas nos mesmos grupos. Ao analisar a m?dia da intensidade de fluoresc?ncia (MFI) encontramos uma express?o diminu?da de CD27 em c?lulas T CD4 nos controles quando comparado as mulheres com c?ncer de mama expostas e n?o expostas a maus tratos (p = 0.002). N?o observamos diferen?as entre os grupos CM+ e CM- em rela??o a sorologia IgG para citomegalov?rus entre os grupos CM + e CM - (p=0.24), entretanto foi presenciado entre os controles e indiv?duos com maus tratos (p= 0.008). Conclus?o: nossos achados sugerem que a presen?a de maus tratos n?o est? diretamente associada com um perfil de imunossenesc?ncia em mulheres rec?m diagnosticada com c?ncer de mama. Entretanto, quando avaliamos mulheres com c?ncer de mama e indiv?duos saud?veis esse perfil senescente torna-se evidente. Logo, torna-se necess?rio a realiza??o de novos estudos para explorar o impacto da presen?a de marcadores de senesc?ncia no tratamento e progn?stico do c?ncer de mama.