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.

Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2018-04-17T11:55:36Z No. of bitstreams: 1 LAURA_ESTEVES_PETERSEN_TES.pdf: 11032751 bytes, checksum: 519545881a34767f1600d5189b0ddc8e (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2018-05-04T17:09:09Z (GMT) No. of bitstreams: 1 LAURA_ESTEVES_PETERSEN_TES.pdf: 11032751 bytes, checksum: 519545881a34767f1600d5189b0ddc8e (MD5) / Made available in DSpace on 2018-05-04T17:15:58Z (GMT). No. of bitstreams: 1 LAURA_ESTEVES_PETERSEN_TES.pdf: 11032751 bytes, checksum: 519545881a34767f1600d5189b0ddc8e (MD5) Previous issue date: 2018-03-14 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Rheumatoid Arthritis (RA) is an autoimmune and inflammatory disease which leads to premature immunosenescence and the development of articular and extra-articular manifestations, including joint damage and cognitive dysfunction, respectively. Peripheral immune system cells and the chronic inflammation, both of great importance for RA, are potentials mechanisms involved in cognitive dysfunction. In contrast, experimental studies have shown the beneficial contribution of immune cells, mainly T cells reactive to central nervous system (CNS) antigens, to neurogenesis and neuroplasticity. Previous data have shown that RA patients besides to have worse performance in cognitive tests, have significantly lower levels of B cells and higher levels of senescent T cells (CD8+CD28-). However, is not known yet which B cells subpopulations are related to poor cognitive performance, if clinical severity of disease (active and controlled disease) impacts on cognition and which factor is responsible for remodeling of peripheral immunity (immunosenescence). Hypotheses about the contribution of latent infections (Cytomegalovirus; CMV), for the development of immunosenescence (observed by telomere shortening and increase of CD28- cells) have been raised. However, it remains in discussion the CMV soroprevalence and its relation with the development of premature immunological senesce in RA. Based on this findings, in this work we have broadly assessment the cognition of RA patients with active and controlled disease, peripheral levels of lymphocytes subsets (T and B cells), neurotrophic factors, cytokines besides that CMV soroprevalence and immunosenescence profile (telomere length and CD28- cells). This work also proposed to explore the relationship between immune mediators, neurotrophic factors and cognitive performance, besides the association between CMV and immunosenescence. For this purpose, 102 RA patients (67 active and 35 controlled disease) and 30 healthy controls adjusted for age, gender and schooling were recruited. The cognitive function, levels of stress and depression were assessment by means of neurocognitive tests (Mini Mental State Examination, Logical memory, digit span, Trail Making Test, N-back, Stroop word-colors) and specific questionnaires (Beck Depression Inventory ?II for depression and Perceived Stress Scale for stress). Twenty ml of blood were collected and, after plasma separation, the peripheral blood mononuclear cells (PBMCs) were isolated by centrifugation gradient. PBMCs were immunophenotyped by flow cytometry to investigate the frequency of T and B cells subsets. The genomic DNA was isolated from PBMCs and the telomere length was evaluated by real time PCR. Neurotrophic factors, cytokines, IgM and IgG anti-CMV were measured in plasma by means of ELISA (NF and CMV) and Citometric Bead Array (CBA; cytokines). In general, patients with active disease had worse cognitive performance, followed by patients with controlled disease producing cells, in addition to high levels of cytokines, with the exception of IL-17A. Higher levels of BDNF were found in patients with active RA followed by controlled disease and control group. The peripheral levels of GDNF were lower in patients with active RA than control group. The IL-6 was predictor of the performance in Trail Making test. IgM and IgG anti-CMV antibody titers did not differ between patients and controls. Only IgG anti-CMV was positively associated with age and senescent cells. In conclusion, RA patients with active disease had worse performance in cognitive tasks that were related to peripheral immune mediators (cells and cytokines). Besides that, we found that late infection (IgG) by CMV were associated only with CD4+CD27-CD28- and haven?t correlated with other immunosenescence characteristics. However, understand in which sense e how the relationship between the peripheral environment and the CNS is established, may contribute to development of preventive interventions to cognitive impairments and premature immunosenescence, since both factors are associated to health and well-being of individuals. / A artrite reumatoide (AR) ? uma doen?a autoimune inflamat?ria que leva ? imunossenesc?ncia prematura e ao desenvolvimento de manifesta??es articulares e extraarticulares, entre elas a destrui??o da articula??o e o decl?nio cognitivo, respectivamente. C?lulas do sistema imune perif?rico e a inflama??o cr?nica, ambos de grande import?ncia para a AR, s?o potenciais mecanismos envolvidos na disfun??o cognitiva. Em contrapartida, estudos experimentais tem revelado a contribui??o ben?fica das c?lulas imunol?gicas, principalmente c?lulas T reativas a ant?genos do sistema nervoso central (SNC), para a neurog?nese e neuroplasticidade. Dados pr?vios apontam que pacientes com AR al?m de apresentarem piores desempenhos nos testes cognitivos, tem significativamente menos c?lulas B e mais c?lulas T com perfil senescente (CD8+CD28-). Entretanto ainda n?o se sabe quais subpopula??es de c?lulas B est?o relacionadas ao pior desempenho cognitivo, se a severidade cl?nica da doen?a (doen?a ativa e controlada) impacta sobre a cogni??o e qual fator seria respons?vel pelo remodelamento da imunidade perif?rica (imunossenesc?ncia). Hip?teses sobre a contribui??o de infec??es latentes, como a causada pelo citomegalov?rus (CMV), para o desenvolvimento da imunossenesc?ncia, observada pelo encurtamento telom?rico e aumento na frequ?ncia de c?lulas CD28-, tem sido levantadas. Por?m, permanece em discuss?o a soropreval?ncia da infec??o pelo CMV e sua real rela??o com o desenvolvimento da senesc?ncia imunol?gica prematura na AR. Com base nestas constata??es, nesta tese n?s avaliamos amplamente a cogni??o de pacientes com AR ativa e controlada, n?veis perif?ricos de subtipos linfocit?rios (c?lulas T e B), fatores neurotr?ficos (FN), citocinas, al?m da soropositividade para CMV e perfil de imunossenesc?ncia prematura (encurtamento telom?rico e aumento de c?lulas CD28-). Esta tese tamb?m se prop?s explorar a rela??o entre mediadores imunes, FN e desempenho cognitivo, e a associa??o entre CMV e caracter?sticas de imunossenesc?ncia. Para esta finalidade, 102 pacientes com AR (67 com doen?a ativa e 35 com doen?a controlada) e 30 controles saud?veis ajustados para idade, g?nero e escolaridade foram recrutados. A fun??o cognitiva, n?veis de estresse e depress?o foram avaliados por meio de testes neurocognitivos (Mini Exame do Estado Mental, Mem?ria L?gica, Subteste de D?gitos, Trail Making Test, N-back, Stroop palavras-cores) e question?rios espec?ficos (Beck Depression Inventory ?II e Escala de Estresse Percebido). Foram coletados 20 ml de sangue e, ap?s a separa??o do plasma, as c?lulas mononucleares do sangue perif?rico (PBMCs) foram isoladas por gradiente de centrifuga??o. PBMCs foram imunofenotipadas por citometria de fluxo para investigar a frequ?ncia de subpopula??es de c?lulas T e B. FN, citocinas, IgM e IgG anti-CMV foram dosados no plasma atrav?s da t?cnica de ELISA (FN e CMV) e Citometric Bead Array (CBA; citocinas). De forma geral, pacientes com doen?a ativa tiveram o pior desempenho nos testes cognitivos, seguido pelos indiv?duos com doen?a controlada e grupo controle. Pacientes com AR tiveram elevados n?veis perif?ricos de c?lulas B imaturas e produtoras de anticorpos, al?m de elevados n?veis das citocinas, com exce??o da IL-17. Maiores concentra??es de BDNF foram observadas nos indiv?duos com AR ativa, seguido pelo grupo controlado e controle. Os n?veis perif?ricos de GDNF foram menores em pacientes com AR ativa do que em indiv?duos controle. A IL-6 apresentou-se como preditora do desempenho do Trail Making Test. T?tulos dos anticorpos IgM e IgG anti-CMV n?o diferiram entre pacientes e controles. Somente o IgG anti-CMV foi relacionado positivamente com idade e c?lulas senescentes. Concluindo, pacientes com AR ativa apresentam pior desempenho em tarefas cognitivas as quais est?o relacionadas a mediadores imunes perif?ricos. Al?m disso, observou-se que infec??es tardias pelo CMV (t?tulos de anticorpos IgG anti-CMV) foram somente associadas a c?lulas T senescentes e n?o se correlacionaram com outras caracter?sticas da imunossenesc?ncia. Portanto, compreender em qual sentido e como a rela??o entre o ambiente perif?rico e do SNC se estabelece, pode contribuir para o desenvolvimento de interven??es preventivas ao d?ficit cognitivo e senesc?ncia prematura, uma vez que ambos fatores est?o associados a sa?de e o bem ? estar dos indiv?duos.

Data de aprova??o ausente. / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-02-13T18:51:06Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_fernandes_araujo_almeida.pdf: 2531556 bytes, checksum: 11a2583c0414dc55d6f907a2566a4de5 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-03-06T12:21:52Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_fernandes_araujo_almeida.pdf: 2531556 bytes, checksum: 11a2583c0414dc55d6f907a2566a4de5 (MD5) / Made available in DSpace on 2017-03-06T12:21:52Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_fernandes_araujo_almeida.pdf: 2531556 bytes, checksum: 11a2583c0414dc55d6f907a2566a4de5 (MD5) Previous issue date: 2016 / O carcinoma espinocelular oral (CECO) ? uma neoplasia maligna que acomete principalmente idosos. O desencadeamento da doen?a ? relacionado ? exposi??o aos fatores de risco e ao decl?nio da fun??o imune associada ao envelhecimento cronol?gico. Alguns estudos t?m demonstrado um aumento na incid?ncia de CECO em adultos jovens, sugerindo envolvimento de outros fatores na etiologia da doen?a, como uma altera??o do sistema imunol?gico. As c?lulas dendr?ticas (CDs) s?o c?lulas apresentadoras de ant?geno profissionais e estimuladoras eficazes para a expans?o clonal de linf?citos. Acredita-se que uma altera??o da fun??o das CDs em pacientes com c?ncer contribui para a falha da resposta antitumoral, levando a uma consequente progress?o da doen?a. O objetivo deste estudo foi avaliar, atrav?s da imunoistoqu?mica, se h? diferen?a na quantifica??o tissular dos subgrupos de CDs, associada ? idade, em esp?cimes de bi?psias de CECO. Para isso, foram selecionados casos de CECO de pacientes em tr?s diferentes faixas et?rias: G1 (< 40 anos de idade, n = 12), G2 (? 40 at? < 60 anos de idade, n = 15) e G3 (? 60 anos de idade, n = 14). Os marcadores utilizados foram S100, CD1a, CD207 (para CDs imaturas, imCDs), CD83 e CD208 (para CDs maduras, mCDs). As imagens foram capturadas dos campos com maior intensidade de marca??o nas l?minas histol?gicas e a quantifica??o celular foi realizada com o aux?lio do software Image J. A frequ?ncia e localiza??o das CDs foram avaliadas e analizadas estatisticamente nas regi?es intratumoral (intertumoral e ou estromal) e extratumoral. No geral, imCDs foram significativamente mais frequentes que mCDs em todos os grupos. ImCDs e mCDs mostraram preferencialmente localiza??o intratumoral e extratumoral respectivamente. Comparando G1 versus G2 / G3 foi observada um significativo menor n?mero de mCDs em G1. Na compara??o de G1 em rela??o a G2 ou G3 houve um n?mero significativamente menor de ambas, imCDs e mCDs. Nossos resultados mostram que existe uma menor quantidade de imCDs e mCDs em CECO afetando pacientes jovens em compara??o com idosos, sugerindo um comprometimento da resposta imune antitumoral em G1e permitindo a progress?o do tumor. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Odontologia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, [2016]. / Oral squamous cell carcinoma (OSCC) is a malignant neoplasm that mainly affects elderly people. The onset of the disease is related with exposure to risk factors and the decline of immune function associated with chronological aging. Previous studies have demonstrated an increased incidence of CECO in young adults, suggesting involvement of other factors in the etiology of the disease, such as a change in the immune system. Dendritic cells (DCs) are professional antigen presenting cells and effective for stimulating clonal expansion of lymphocytes. It is believed that a change in the function of DC in cancer patients contributes to the failure of the anti-tumor response, leading to a consequent progression of the disease. The aim of this study was to evaluate, by immunohistochemistry, if there were differences in tissue quantification of DCs subsets, associated with age in specimens of OSCC biopsies. For this OSCC cases were selected at three different ages: G1 (< 40 years old, n = 12), G2 (? 40 to <60 years old, n = 15) and G3 (? 60 years old, n = 14). The markers used were S100, CD1a, CD207 (for immature DCs, imDCs), CD83 and CD208 (for mature DCs, mDCs). Images were captured from fields with higher intensity staining in histological sections and cell quantification was performed with Image J software help. The frequency and localization of immunostained DCs were analyzed in intratumoral (intranestal and/or extranestal) and extratumoral areas and statistically compared. Overall, imDCs than mDCs were significantly more frequent in all groups. ImDCs and mDCs showed preferential intratumoral and extratumoral localization, respectivel. Comparing G1 than G2/G3 showed a significant lesser number of mDCs. G1 versus G2 or G3 there is a significantly lower number of both imDCs and mDCs. Our results show a lower number of both imDCs and mDCs in OSCC affecting younger than elderly patients, suggesting impairment of an effective antitumor immune response in G1 and enabling tumor progression, showing a gradual establishment of the antitumor immune response mediated by DCs according to the age, but with defects in quality.