Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 114-146).

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 112-132).

Diss. (sammanfattning) Linköping : Linköpings universitet, 2010. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.

Made available in DSpace on 2015-07-22T20:50:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-29. Added 1 bitstream(s) on 2015-08-11T03:25:40Z : No. of bitstreams: 1 Publico-062a.pdf: 992519 bytes, checksum: 41750ff4e6e38ad6517d6c7541049844 (MD5). Added 1 bitstream(s) on 2015-08-11T03:25:40Z : No. of bitstreams: 2 Publico-062a.pdf: 992519 bytes, checksum: 41750ff4e6e38ad6517d6c7541049844 (MD5) Publico-062b.pdf: 1669897 bytes, checksum: 5ca81b8b1f9a75f45902de9ce4bc36f7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O GB vírus C (GBV-C) está constituído por uma fita única de RNA de polaridade positiva e pertence à família Flaviviridae. Possui uma seqüência e organização genómica parecida ao vírus da hepatite C, (HCV). A infecção pelo GBV-C não foi associada a nenhuma patologia, embora, na co infecção com o HIV, tenha sido associada a uma sobrevida maior e retardo no desenvolvimento da imunodeficiência. O efeito benéfico do GBV-C parece ser mediado por alterações na resposta imune celular; contudo, os possíveis mecanismos para explicar esse efeito ainda não foram esclarecidos. Neste trabalho investigamos a freqüência e características genotípicas assim como o impacto da infecção pelo GBV-C nos indivíduos infectados pelo HIV-1. No primeiro manuscrito examinamos os conhecimentos descritos na literatura referentes à coinfecção e propusemos algumas hipóteses para explicar esses efeitos. Posteriormente, descrevemos a taxa de infecção, a prevalência, incidência e características genotípicas do GBV-C nesta população. Assim, uma considerável freqüência de infecção pelo GBV-C foi observada e a análise filogenética dos isolados de GBV-C mostraram ser do genótipo 1 e 2. Foi observada também uma correlação inversa entre a carga viral do GBV-C e a carga viral do HIV na inclusão e um ano depois, assim como uma correlação positiva, mas não significativa, entre a carga viral do GBV-C e a contagem de linfócitos T CD4+. Finalmente, avaliamos o efeito da viremia pelo GBV-C na ativação celular em recém infectados pelo HIV-1. Os pacientes foram agrupados em GBV-C viremicos e não virémicos e foram avaliados para a contagem de linfócitos T, marcadores de ativação celular e carga viral do GBV-C e HIV-1. Foram realizadas análises de univariada e multivariada para identificar variáveis associadas com ativação celular. Demonstramos que a viremia pelo GBV-C foi correlacionada com uma diminuição da ativação celular nos indivíduos HIV positivos e este efeito mostrou se independente da carga viral do HIV. Assim, esta associação entre a replicação do GBV-C e menor ativação celular pode explicar, pelo menos em parte, a proteção conferida pelo GBV-C na progressão da doença nos indivíduos infectados pelo HIV-1. / GB virus C (GBV-C) is a single stranded positive sense RNA virus, which is a member of the Flaviviridae. It has a close sequence homology and genomic organization to hepatitis C virus (HCV). No disease has been associated with GBV-C infection but coinfection with human immunodeficiency virus (HIV) leads to improved morbidity and mortality for the HIV infected subjects. The mechanism of the beneficial effect of GBV-C appears to be mediated by alterations in the cellular immune response. In this study we investigated the frequency and genotyping characteristics as well as the impact of the GBV-C infection among recently HIV-1 infected individuals. In the first manuscript we examined the current knowledge concerning this co-infection and developed hypotheses to explain its effects. Subsequently, we described the rate of infection, the prevalence, incidence and genotypic GBV-C characteristics in this population. In that regard, a considerable frequency of GBV-C infection was observed and the phylogenetic analysis of the GBVC isolates revealed the predominance of genotypes 1 and 2. Also, it was observed an inverse correlation between GBV-C load and HIV-1 load at the enrollment and after one year of follow-up, and a positive, but not statistically significant, correlation between GBV-C load and CD4+ T lymphocyte counts. Finally, we have investigated the effect of GBV-C viremia on T cell activation in early HIV-1-infection. The volunteers were enrolled into two groups: GBV-C viremic and non viremic, all co-infected with HIV-1. They were evaluated for T cell counts, cellular activation markers, GBV-C RNA detection, and HIV-1 viral load. Non-parametric univariate and multivariate analyses were carried out to identify the variables associated with cellular activation. We demonstrated that the GBV-C viremia is correlated with a lower T cell activation in HIV-1-infected individuals and this effect was independent of HIV-1 viral load. The association between GBV-C replication and lower T-cell activation may explain, at least in part, the protection conferred by this virus against disease progression to immunodeficiency in HIV-1-infected patients. / TEDE / BV UNIFESP: Teses e dissertações

L’apprêtement antigénique par les protéases intracellulaires et la présentation des épitopes sont essentiels pour la reconnaissance des cellules infectées par les lymphocytes CD8+. Ici nous avons montré que certains inhibiteurs de la protéase de la VIH (IPs) modulent l’activité de la protéasome et aminopeptidase impliqué dans l’apprêtement antigénique endogène et l’activité cathepsins importante dans l’apprêtement croisée. Deux IPs agissent directement sur les cathepsins et leurs régulateurs en inhibant les activités kinase, NOX2 et en régulant le pH phagolysosomal. Les IPs ont changé la dégradation des protéines viral et la production des épitopes de façon séquence- et cellule-spécifique, ont altéré la présentation direct et croisée des épitopes, et ont partiellement changé l’auto-peptidome des cellules primaires. La modulation par les drogues de l’apprêtement et la présentation des épitopes peut fournir une approche thérapeutique alternative pour moduler la reconnaissance immunitaire. / Antigen processing by intracellular proteases and peptidases and epitope presentation are critical for recognition of pathogen-infected cells by CD8+ T lymphocytes. Here we show that several HIV protease inhibitors (PIs) prescribed to HIV-infected persons variably modulate proteasome and aminopeptidase activities involved in endogenous antigen presentation and cathepsin activities involved in antigen cross-presentation. Two HIV PIs acted directly on cathepsins and on their regulators by inhibiting kinases, NOX2 and the regulation of phagolysosomal pH, subsequently enhancing cathepsin activities. HIV PIs modified HIV protein degradation and epitope production in a sequence- and cell-dependent manner, altered direct- and cross-presentation and T cell-mediated killing, and partly changed the self-peptidome of primary cells. Drug-induced modulation of antigen processing and peptidome may provide an alternate therapeutic approach to modulate immune recognition.

<p>UVOD: Glavni problem u lečenju karcinoma dojke je kako na osnovu kliničke klasifikacije i morfolo&scaron;kih osobina tumora predvideti njegovo dalje pona&scaron;anje. Vrlo često ni kombinacija standardnih prognostičkih faktora ne daje odgovor o potrebi davanja adjuvantne hemioterapije. U cilju sprovođenja adekvatne dalje terapije karcinoma dojke i otkrivanja agresivnih tipova tumora, a nakon hirur&scaron;kog lečenja, postoji stalna potreba za pronalaženjem novih pokazatelja pomoću kojih bi se identifikovale bolesnice koje imaju povećan rizik od razvoja relapsa bolesti. CILJEVI: Ciljevi su bili da se utvrdi prisustvo, lokalizacija i distribucija tumor infiltri&scaron;ućih limfocita, kako ukupnih, tako i CD4+ i CD8+ T limfocita u tumoru dojke, njihova povezanost sa standardnim prognostičkim parametrima, kao i njihov prognostički značaj tj. razlike u nivou infiltracije limfocita u tumoru u odnosu na pojavu relapsa bolesti i dužinu preživljavanja. METOD: Istraživanjem je obuhvaćeno 120 bolesnica sa invazivnim duktalnim karcinomom, sa tumorom lokalizovanim samo u dojci, bez zahvatanja kože i grudnog mi&scaron;ića, sa veličinom tumora do 5 cm, bez udaljenih visceralnih i ko&scaron;tanih metastaza, koje su operisane u Institutu za onkologiju Vojvodine. U istraživanje su uključene bolesnice bez metastaza u limfnim čvorovima pazu&scaron;ne jame i bolesnice sa metastazama u limfnim čvorovima pazu&scaron;ne jame. Istraživanjem nisu obuhvaćene bolesnice koje su primale neoadjuvantnu (preoperativnu) hemioterapiju, kao ni bolesnice sa multifokalnim i multicentričnim tumorima. REZULTATI: Gust limfocitni infiltrat uočen je u 14% tumora dojke, umeren limfocitni infiltrat uočen je u 38%, a oskudan u 43% tumora dojke. Limfocitni infiltrat nije uočen u 5% tumora. Gust infiltrat CD4+ limfocita uočen je u 8% tumora dojke, umeren u 44%, a oskudan u 43% tumora dojke. CD4+ limfociti nisu uočeni u 5% tumora. Gust infiltrat CD8+ limfocita uočen je u 1% tumora dojke, umeren u 23%, a oskudan u 66% tumora dojke. CD8+ limfociti nisu uočeni u 10% tumora. Utvrđena je pozitivna povezanost između nivoa TIL-a i CD4+ limfocita i veličine tumora, histolo&scaron;kog stepena diferentovanosti tumora, prisustva metastaza u limfnim čvorovima pazu&scaron;ne jame, HER-2 statusa, tripl negativnog tumora i relapsa bolesti. Utvrđena je negativna povezanost između nivoa TIL-a i CD4+ limfocita i estrogen i progesteron receptora, kao i godina starosti. Utvrđena je pozitivna povezanost između nivoa CD8+ limfocita i histolo&scaron;kog gradusa tumora, kao i HER-2 statusa. Utvrđena je negativna povezanost između nivoa CD8+ limfocita i estrogen i progesteron receptora, kao i godina starosti. ZAKLJUČAK:Rezultati ovog istraživanja pokazuju povezanost tumor infiltri&scaron;ućih limfocita i CD4+ limfocita sa brojnim negativnim prognostičkim faktorima, te kraćim vremenom slobodnog intervala bez bolesti, &scaron;to sve ukazuje na to da su tumor infiltri&scaron;ući limfociti kao i CD4+ limfociti lo&scaron; prognostički faktor kod bolesnica sa rakom dojke.</p> / <p>INTRODUCTION: The main problem in the treatment of breast cancer that based on clinical classification and morphological characteristics of the tumor to predict its future behavior. Very often, not a combination of standard prognostic factors does not answer the need of giving adjuvant chemotherapy. In order to implement adequate further treatment of breast cancer and detection aggressive types of tumor, after surgical treatment, there is a constant need to find new indicators by which we can identify patients who have an increased risk of relapse. OBJECTIVES: The objectives were to determine the presence, localization and distribution of tumor infiltrating lymphocytes, in total, as well as CD4+ and CD8+ T lymphocytes in breast cancer, their correlation with standard prognostic parameters, as well as their prognostic value i.e. differences in the level of infiltration of lymphocytes in a tumor in relation to the occurrence of disease relapse and survival. METHOD: The study included 120 patients with invasive ductal carcinoma, tumor localized only in the breast without involvement of the skin and pectoral muscle, the size of tumors up to 5 cm without distant visceral and bone metastases, which are operated at the Institute of Oncology. The study included patients without metastases in axillary lymph nodes and patients with metastases in axillary lymph nodes. The research not covered by patients receiving neoadjuvant chemotherapy, or patients with multifocal and multicentric tumors. RESULTS: The high amount of lymphocytic infiltrate was observed in the 14% a breast tumor, a moderate amount of lymphocytic infiltrate was observed in 38%, and the low in 43% breast tumors. Lymphocytic infiltrate was not observed in 5% of the tumor. High CD4+ lymphocyte infiltration was observed in 8% of breast, moderate in 44%, and the low in 43% of breast tumors. CD4+ lymphocytes were not observed in 5% tumors. High infiltration of CD8+ lymphocytes was observed in 1% of breast, moderate in 23%, and the low 66% breast tumors. CD8+ lymphocytes have not been observed in 10% tumors. There is a positive correlation between the level of TIL and CD4+ lymphocytes and tumor size, histological grade of tumor differentiation, presence of metastases in axillary lymph nodes, HER-2 status, triple negative tumors and relapses of disease. There was a negative correlation between the level of TIL and CD4+ cell counts and estrogen and progesterone receptors, as well as age. There is a positive correlation between the level of CD8+ cells and histological grade of the tumor, and HER-2 status. There was a negative correlation between the level of CD8+ lymphocytes and estrogen and progesterone receptors, as well as age. CONCLUSION: The results of this study demonstrate the association between tumor infiltrating lymphocytes and CD4+ lymphocytes with a number of negative prognostic factors, and shorter free interval without the disease, all of which indicates that the tumor infiltrating lymphocytes and CD4+ lymphocytes bad prognostic factor in patients with breast cancer.</p>