Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-04-17T18:56:08Z No. of bitstreams: 1 Ribeiro, Mara Fernandes [Dissertação, 2014].pdf: 3051138 bytes, checksum: 7536143779553e35c3e2d8ad11a9b9bf (MD5) / Made available in DSpace on 2017-04-17T18:56:08Z (GMT). No. of bitstreams: 1 Ribeiro, Mara Fernandes [Dissertação, 2014].pdf: 3051138 bytes, checksum: 7536143779553e35c3e2d8ad11a9b9bf (MD5) / Os acidentes causados por aranhas do gênero Loxosceles representam importante problema de saúde pública no Brasil, sendo as principais espécies de importância médica L. intermedia, L. laeta e L. gaucho. O veneno dessas aranhas promove grave dermonecrose no local da picada, e menos comumente, doença sistêmica que pode ser fatal. O mecanismo de ação desse veneno não está completamente elucidado, trata-se de um processo multifatorial, que envolve a ação direta do veneno sobre os tecidos e a resposta do organismo a agressão causada pelo mesmo. Os camundongos constituem o modelo experimental menos susceptível ao desenvolvimento dos efeitos locais decorrentes do envenenamento por aranhas Loxosceles, dessa forma, sua utilização representa grande interesse clínico, cujo objetivo é desvendar tal mecanismo de proteção presente nestes animais. Este trabalho teve como objetivo caracterizar as atividades do veneno de Loxosceles intermedia, bem como avaliar as atividades in vivo deste veneno em camundongos. A manipulação e os procedimentos com os animais obedeceram aos princípios da CEUA/UFF (Comissão de Ética no Uso de Animais da Universidade Federal Fluminense). Foi demonstrado, in vitro, que o veneno de L. intermedia não apresenta atividade fosfolipásica A2, a atividade hialuronidásica e colágenásica foram dependente da concentração do veneno enquanto que a atividade proteolítica e esfingomielinásica foram observadas apenas em altas concentrações. Para descrever as ações do veneno de L. intermedia em camundongos, foi proposta a utilização de três diferentes linhagens: BALB/c, C57BL/6 e Suiço. A atividade edematogênica na pata dos camundongos inoculados com o veneno foi observada para as três linhagens testadas, sendo persistente por 24 horas, apenas para as linhagens BALB/c e C57BL/6. A análise histopatológica do local de inoculação intradérmica do veneno no abdomen apresentou diferenças relevantes, como, intensa congestão vascular em Suiços e presença de infiltrado inflamatório no local de inoculação na pele de BALB/c e C57BL/6. A partir destes achados, investigou-se a mobilização de células inflamatórias a partir da medula óssea, no baço e no sangue por citometria de fluxo, que demonstrou resposta imunológica inata típica, com aumento da cinética de células mieloides e linfócitos T citotóxicos para camundongos C57BL/6, e resposta tipicamente adquirida/humoral, com aumento preferencial de linfócitos B convencionais e T auxiliar para camundongos BALB/c. Desta forma, este trabalho demonstrou que modelos animais semelhantes podem apresentar diferentes respostas a inoculação deste veneno. A presença do infiltrado inflamatório no local de inoculação do veneno e a mobilização de células inflamatórias a partir da medula óssea, baço e sangue revelaram que diferentes linhagens de camundongos apresentam diferenças no tipo celular envolvido na resposta imunológica decorrente do envenenamento ou esta diferença pode estar relacionada ao tempo e velocidade da resposta em cada linhagem de camundongos. A partir destes resultados este trabalho sugere que camundongos da linhagem BALB/c podem ser utilizados como modelo para estudar a produção de IgM e IgG induzido pelo veneno, incluindo análise de citocinas, quimiocinas e mecanismos moleculares, por outro lado camundongos C57BL/6 podem ser utilizados para descrever a participação de células mielóides durante o envenenamento por aranhas do gênero Loxosceles / Accidents caused by spiders of the genus Loxosceles represent an important public health problem in Brazil, being the major species of medical importance L. intermedia, L. laeta and L. gaucho. The venom of these spiders induces an intense dermonecrosis at the bite site, and less commonly, systemic disease that can be fatal. The mechanism of action of this venom is not fully elucidated, it is a multifactorial process, which involves the direct action of the venom on the tissues and the body's response to aggression caused by it. The mice are an experimental model less susceptible to development the local effects of poisoning Loxosceles spiders. Thus, their use is great clinical interest, whose goal is to unravel the mechanism of this protection in these animals. This study aimed to characterize the activities of Loxosceles intermedia venom, as well as evaluating the in vivo activity of this venom in mice. Manipulation and procedures with animals obeyed the principles of CEUA / UFF (Ethics Committee on Animal Use Universidade Federal Fluminense). It has been shown in vitro that the venom of L. intermedia shows no phospholipase A2 activity and the hyaluronidase and collagenase were dependent on the concentration of the poison and while the proteolytic and sphingomyelinase activity were observed only at high concentrations. To describe the actions of the venom of L. intermedia in mice, it was proposed to use three different strains: BALB/c, C57BL/6 and Swiss. The activity in the paw edema of mice inoculated with the venom was observed for the three strains tested, being persistent for 24 hours, only for the strains BALB/c and C57BL/6. Histopathological analysis of the site of venom intradermal inoculation in the abdomen showed significant differences, as intense vascular congestion in Swiss and inflammatory infiltration at the site of inoculation in the skin of BALB/c and C57BL/6. From these findings, we investigated the mobilization of inflammatory cells from the bone marrow, spleen like effector organ, and migration into the blood by flow cytometry, which showed a typical innate immune response, with increased kinetics of myeloid cells and cytotoxic T lymphocytes to C57BL/6 mice, and response typically acquired / humoral, with a preferential increase of conventional B lymphocytes and T helper to BALB/c mice. Thus, this study demonstrated that similar animal models may have different responses to inoculation of this venom. The presence of the inflammatory infiltrate at the site of inoculation of the venom and the mobilization of inflammatory cells from the bone marrow, spleen and blood revealed that different strains of mice differ in cell type involved in the immune response resulting from poisoning and this difference can be related time and speed of response for each strain of mice. From these results, this study suggests that BALB/c mice can be used as a model to study the production of IgM and IgG induced by the venom, including analysis of cytokines, chemokines and molecular mechanisms, on the other hand C57BL/6 mice can be used to describe the involvement of myeloid cells during poisoning by spiders of the genus Loxosceles

Magister Chirurgiae Dentium (MChD) / Oral lichen planus (OLP) is a T-cell mediated chronic inflammatory disease with different clinical types that remains inscrutable in respect of its pathogenetic mechanisms and effective therapy. Increased apoptosis may influence the histopathological criteria of oral lichen planus (decrease in thickness of the epithelium and band of inflammatory infiltrate). Null hypothesis: The apoptotic rate does not correlate with a decrease in the epithelial thickness as well as the thickness of the band of inflammatory infiltrate in OLP. Aim: The present study aims to quantify apoptotic activity and to correlate the apoptotic rate with epithelial thickness as well as thickness of the inflammatory infiltrate of OLP cases diagnosed at Tygerberg Hospital from 2006 – 2015. Further, the epithelial thickness and thickness of the inflammatory infiltrate were also assessed for their association, if any. Materials and Methods: The study sample comprised 17 diagnostically verified cases of OLP. Sections stained with Haematoxylin and Eosin (H&E) were used to identify and count the number of apoptotic cells as well as measure the thickness of epithelium and the thickness of the lymphocytic inflammatory infiltrate by using software morphometric analysis (Zen Blue lite 2012). Statistical analysis was applied to analyse the correlation between apoptotic cells and histopathological features of OLP. Results: The present study's results showed no statistically significant association between the apoptotic rate, the epithelial thickness and the thickness of the lymphocytic inflammatory infiltrate.

Les métastases hépatiques compliquent l'évolution de 50% des cancers colorectaux (CCR). Plus de la moitié des patients présentent une récidive à distance avec métastases occultes pour lesquelles une chirurgie peut être réalisée dans moins de 20% des cas. L'ablation par radiofréquence (RFA) induit une réponse lymphocytaire T qui n'est pas évaluée après une intervention chirurgicale seule. L'immunothérapie combinée à la RFA pourrait potentialiser cet effet conduisant à une réponse tumorale à distance. Nous proposons une approche qui combine la RFA avec hydrogel thermoreversible libérant des agents immunomodulateurs (GMCSF et BCG) sur le site du traitementPremièrement, nous nous sommes intéressés à la sélection et à la caractérisation de la formulation optimale d’hydrogel par des techniques physicochimiques. Les propriétés de l'hydrogel ont été étudiées par rhéologie et des tests de muco-adhésion ont été mis en place. Le temps de résidence de l'hydrogel et de la protéine dans la zone tumorale a été démontré par imagerie optique. De plus, la cinétique de libération et l'intégrité du GMCSF encapsulé ont été déterminées. Ensuite, nous avons démontré l’efficacité de l’association de la RFA avec le dépôt local de l’hydrogel immunomodulatuer sur un modèle murin de cancer colorectal. En effet, nous avons observé une survie améliorée des animaux et régression complète des tumeurs distantes chez les animaux traités par la combinaison complète. Cette réponse est caractérisée par un niveau élevé de sécrétion de cytokines pro-inflammatoires par les cellules T CD4 et TCD8 et une augmentation de l’infiltrat lymphocytaire dans les tumeurs. Ceci a permis d'envisager une association avec l'immunothérapie anti-PD1 dans le traitement de macrométastases échappant au traitement combiné RFA avec l’hydrogel immunomodulateur. En effet, l’immunothérapie dans le traitement du cancer colorectal métastatique présente une efficacité limitée chez les patients. Notre travail propose a démontré que l’efficacité de l’immunomodulation locale dans l’amélioration des réponses immunitaires dans le cancer colorectal. Ces résultats permettent de reconsidérer l’utilisation de l’immunothérapie chez les patients atteints de CCR métastatique non MSI. / Liver metastases complicate the progression of 50% of colorectal cancers (CRC). More than half of the patients have recurrent remissions with occult metastases for which surgery can be performed in less than 20% of cases. Radiofrequency ablation (RFA) induces a T lymphocyte response that is not observed after surgery alone. Combined immunotherapy with RFA may potentiate this effect leading to a distant tumor response. We propose an approach that combines RFA with thermoreversible hydrogel releasing immunomodulatory agents (GMCSF and BCG) at the treatment site.First, we focused on the selection and characterization of the optimal hydrogel formulation by physicochemical techniques. The properties of the hydrogel were studied by rheology and mucoadhesion tests were set up. The residence time of the hydrogel and the protein in the tumor zone was demonstrated by optical imaging. In addition, the release kinetics and integrity of the encapsulated GMCSF were determined. Then, we demonstrated the effectiveness of the combination of RFA with the local deposition of the immunomodulatory hydrogel on a mouse model of colorectal cancer. Indeed, we observed improved survival of animals and complete regression of distant tumors the complete treatment group. This response is characterized by a high level of pro-inflammatory cytokine secreted by CD4 and TCD8 T cells and an increase Lymphocytes infiltrating tumors. The immune escape of large lesions was reversed by association with anti-PD1 immunotherapy Indeed, immunotherapy in the treatment of metastatic colorectal cancer has limited efficacy in patients. Our work has demonstrated the effectiveness of local immunomodulation in improving immune responses in colorectal cancer. These results make it possible to reconsider the use of immunotherapy in patients with non-MSI metastatic CRC.

Thesis (MBA (Business Management))--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Soon after the development of the internet as a network structure connecting computers on a global scale, was the introduction of malicious computer code, which was disseminated through this network. Initially this code was the relegation of pranksters, but evolved quickly to be code causing destruction, intrusion and loss of privacy while on the internet. This code became known as the computer virus and was soon used by fraudsters to infiltrate networks to create deception and fraud for financial gain. It has become of paramount importance for users of the internet to protect themselves and their networks from these attacks, through various ingenious mechanisms of protection. The traditional mainstay for computer virus protection has been the software approach using counter code to protect against any malicious computer code. This protection has had limited success as the very nature of malicious code is constantly changing and evolving, making it sometimes an impossible task for internet users to be protected with the latest anti-virus software for protection. The author of this study introduces a managed anti-virus protection alternative which is delivered by a computer hardware device. This is a new technology and a full description is made of the role of this product as a new product development. The empirical research of this paper focuses around the test for a need for the product described to the point, but excluding commercialization. / AFRIKAANSE OPSOMMING: Kort na die ontwikkeling van die internet as 'n netwerkstruktuur wat rekenaars op 'n globale skaal verbind, was daar die bekendstelling van kwaadwillige rekenaarkodes wat reg deur die netwerk versprei het. Aanvanklik was die kode gemik op die verdrywing van poetsbakkers, maar het spoedig ontwikkel in kodeverdrywing, inmenging en verlies aan privaatheid op die internet. Hierdie kode het bekend geword as die rekenaarvirus, en is spoedig deur bedrieërs gebruik om netwerke te infiltreer om gebruikers te mislei en te bedrieg vir eie finansiële gewin. Dit het vir gebruikers van die internet van uiterste belang geword om hulle en hulle netwerkte teen hierdie aanvalle te beskerm, en wel deur middel van verskeie meganismes. Die bekendste bekermingsmatreël teen die virus is die aanwending van sagteware as teenkode. Hierdie bekermingsmetode het egter tot dusver beperkte sukses behaal, aangesien die aard van kwaadwilligheid voortdurend verander en ontwikkel, sodat dit soms onmoontlik is dat gebruikers deur die nuutste anti-virussagteware beskerm kan word. Die skrywer van hierdie verhandeling stel 'n werkbare, alternatiewe anti-virusbeskermer bekend wat deur rekenaar-hardewareplan daargestel is. Dit het nuwe tegnologie, en 'n volledige beskrywing word gegee van die rol van hierdie produk as 'n nuwe ontwikkeling. Die empiriese navorsing van die verhandeling fokus op die toets vir die noodsaaklikheid van so 'n produk, met die uisluiting van kommersialisering.

Le cancer colorectal est une affection multifactorielle qui conjugue des anomalies épigénétiques et des altérations moléculaires à une déstabilisation du génome. En dehors de deux processus bien établis, l’instabilité des microsatellites et l’instabilité des chromosomes, la déstabilisation du génome peut être consécutive à la réexpression d’éléments génétiques mobiles, HERV et LINEs-­‐1. Les HERVs ont un potentiel de modulation de l’immunité qui pourrait contrôler les réactions de l’hôte dans le processus de la carcinogenèse. Compte tenu du rôle que pourrait jouer les HERV, nous avons voulu déterminer la relation entre l’infiltrat lymphocytaire intratumoral et les différentes caractéristiques moléculaires des tumeurs. L’infiltrat lymphocytaire intratumoral a été évalué pour trois marqueurs (CD3, CD8 et CD45RO). Les marqueurs moléculaires analysés ont été : les mutations des gènes B-­‐raf, k-­‐ras et Pi3K, le statut des microsatellites, la méthylation de MLH1 et des LINEs-­‐1, un polymorphisme de C-­‐met et de HLA-­‐G. Ces marqueurs ont été comparés à la prévalence de la délétion du locus Apobec 3 pour une cohorte de 135 patients porteurs de cancer colorectal. / Colorectal cancer is a multifactorial disease associated to epigenetic abnormalities and molecular alterations inducing a destabilization of the cellular genome. Besides two well-­established processes, microsatellite instability and chromosome instability, the destabilization of the genome may follow mobilisation of endongenised mobile genetic elements, such as HERV or LINEs-­‐1. HERV have a potential to modulate immune response, which suggests that they could contribute to the process of carcinogenesis. We thus wanted to determine the relationship between intratumoral lymphocytic infiltration and the different molecular characteristics of tumours, taking into account factors that could influence the expression of HERV, such as the frequently observed deletion of a locus on 22q13 containing the Apobec 3 gene family. The intratumoral lymphocytic infiltration was assessed for three markers (CD3, CD8 and CD45RO). The molecular markers analysed were: the mutations of B-­‐raf, K-­‐ras and Pi3K genes, the instability status of microsatellites, the methylation of MLH1 and LINEs-­‐1, a polymorphism in C-­‐met and in HLA-­‐G. These markers were linked to the deletion of the locus containing Apobec 3 genes, in a cohort of 135 colorectal cancer patients.

Changes in the microenvironment have been implicated in the transition of pre-invasive ductal carcinoma in-situ (DCIS) to invasive breast cancer. Normal myoepithelial cells have a tumour suppressor phenotype but they are altered in DCIS and ultimately lost with transition to invasive cancer. A consistent change in DCIS is up-regulation of the integrin αvβ6 in myoepithelial cells. Preliminary observations identified a correlation between myopeithelial αvβ6 and an increased peri-ductal inflammatory infiltrate. The hypothesis of this study is that the altered myoepithelial phenotype influences the peri-ductal inflammatory environment, which in turn mediates a pro-apoptotic effect on myoepithelial cells contributing to their loss. To investigate this, the inflammatory infiltrate was characterised in a series of DCIS tissue in relation to αvβ6 status. This demonstrated significantly higher levels of CD4+ve and FOXP3+ve T cells around αvβ6+ve DCIS ducts compared to αvβ6-ve ducts (P=<0.01), suggesting an increase in Treg cells. In-vivo, Matrigel plugs containing injected into the flanks of female C57/Blk6 normal mice generated influx of higher levels of CD4+ve cells (p=0.005) and FOXP3+ T cells (p=0.007) in the presence of αvβ6+ve myoepithelial cells compared to αvβ6-ve cells, supporting the findings in human tissue samples. Since Treg cells produce TRAIL that can induce apoptosis, we investigated the influence of αvβ6 on myoepithelial cells on the levels of TRAIL in T cells and the hypothesis that αvβ6-positive myoepithelial ells may be more susceptible to TRAIL-induced apoptosis, leading to loss of the myoepithelial barrier. Firstly, levels of TRAIL in Jurkat and primary T cell populations co-cultured with β4 (ii) or β6 myoepithelial cells were measured. This demonstrated a higher level of TRAIL in primary T cells co-cultured β6 myoepithelial cells compared to those co-cultured with β4 myoepithelial cells. β6+ve and β6-ve myoepithelial cells were exposed to TRAIL, and this demonstrated that TRAIL enhanced apoptosis, measured by cleaved PARP, in β6+ve cells. Furthermore, these cells showed loss of the anti-apoptotic protein Galectin-7, and knockdown of Galectin-7 in normal β6-ve myoepithelial cells rendered them more susceptible to TRAIL-induced apoptosis. In DCIS tissues, an inverse relationship between αvβ6 and Galectin-7 in myoepithelial cells was demonstrated, and Cytokine Array analysis showed that αvβ6+ve myoepithelial cells express higher levels of IL-16, which has a role in Treg cell recruitment. Taken together these results suggest that expression of αvβ6 by myoepithelial cells in DCIS generates a tumour-promoter peri-ductal inflammatory infiltrate through altered cytokine release, is associated with reduced galectin-7 expression and enhances myoepithelial cell apoptosis in response to TRAIL. This provides a potential mechanism by which myoepithelial cells may be lost during evolution of DCIS and so contribute to progression to invasive disease.

Submitted by Adriana Martinez (amartinez@unoeste.br) on 2017-02-10T16:34:06Z No. of bitstreams: 1 Fabíola Azevedo.pdf: 1121993 bytes, checksum: 7e7e5309d63e8985debf7b6b2d17469c (MD5) / Made available in DSpace on 2017-02-10T16:34:06Z (GMT). No. of bitstreams: 1 Fabíola Azevedo.pdf: 1121993 bytes, checksum: 7e7e5309d63e8985debf7b6b2d17469c (MD5) Previous issue date: 2016-12-15 / Introduction: Increased sugarcane production among the rise in use of herbicides, such as the 2,4-dichlorophenoxyacetic acid (2,4-D), might causes several harm for health. Studies that evaluate the alterations due to the use of this herbicide in the hemogram and in the cells of the nasal mucosa are very important, since the information provided helps in understanding the interactions of substances as the 2,4-D with the human organism. Objective: To analyze blood count components and inflammatory infiltrate in experimental models submitted to nebulization with 2,4-D herbicide, in different time intervals and concentrations. Method: Eighty male Swiss mice were divided into four groups (n = 20): saline, low (3,71 x 10-3 grams of active ingredient per hectare), medium (6,19 x 10-3 grams of active ingredient per hectare) and high concentrations (9,28 x 10-3 grams of active ingredient per hectare). All animals were exposed to the nebulization predefined for each group for 15 minutes during different time intervals: 24, 48, 72 and 192 hours. Blood samples were taken by intracardiac puncture and after euthanasia, the muzzle of each animal was removed for mast cell quantification through toluidine blue staining and analysis of the inflammatory infiltrate by Hematoxylin-Eosin staining (HE). Analysis of normality was conduced using the statistical package Shapiro-Wilk, and for statistical analysis of data Two-way Anova test was performed, considering statistically significant results with p <0.05. Results: All animals in the low concentration group presented microcytosis and hypochromia within 72 hours; Eosinophilia and lymphocytosis within 48 hours. Neutrophilia was observed in the high concentration group over the 192 hour interval. The mast cell count showed significantly increased values in the high concentration group, in the time interval of 48 hours, when compared to the 24 hour period. Conclusion: In conclusion, we observed that in spite of changes in haematological values, the 2,4-dichlorophenoxyacetic acid herbicide can not be considered dose-dependent and time-dependent herbicide, but it might provoke allergic reactions. / Introdução: O aumento da produção de cana-de-açúcar em conjunto com a utilização de herbicidas, como o ácido 2,4-diclorofenoxiacético (2,4-D) traz diversos agravos à saúde humana. Estudos que avaliam as alterações decorrentes do uso desse herbicida no hemograma e nas células da mucosa nasal são de grande importância, pois as informações fornecidas podem ser utilizadas com o intuito de ajudar na compreensão da interação dessa substância com o organismo humano. Objetivo: Analisar os componentes do sangue e infiltrado inflamatório em modelos experimentais submetidos à nebulização ao herbicida 2,4-D, em diferentes intervalos de tempo e concentrações. Metodologia: Foram utilizados 80 camundongos Swiss machos, divididos em quatro grupos (n=20): salina, baixa (3,71 x 10-3 gramas de ingrediente ativo por hectare), média (6,19 x 10-3 gramas de ingrediente ativo por hectare) e alta concentração (9,28 x 10-3 gramas de ingrediente ativo por hectare). Todos os animais foram expostos às nebulizações preconizadas para cada grupo por 15 minutos, em diferentes intervalos de tempo: 24, 48, 72 e 192 horas. A coleta das amostras de sangue foi através de punção intracardíaca e após a eutanásia, o focinho de cada animal foi retirado para a quantificação de mastócitos, através da coloração de azul de toluidina e análise do infiltrado inflamatório, pela coloração de Hematoxilina-Eosina (HE). Para a análise de normalidade foi utilizado o pacote estatístico Shapiro-Wilk e para a análise estatítica dos dados, utilizamos o teste Two-way Anova, considerando estatisticamente diferentes os resultados que apresentaram p< 0,05. Resultados: Todos os animais do grupo baixa concentração apresentaram microcitose e hipocromia no intervalo de 72 horas; eosinofilia e linfocitose no intervalo de 48 horas. Foi verificada neutrofilia no grupo alta concentração no intervalo de 192 horas. A contagem de mastócitos apresentou valores significativamente aumentados no grupo alta concentração, no intervalo de tempo de 48 horas, quando comparado ao de 24 horas. Conclusão: Em conclusão observamos que, apesar das alterações nos valores hematológicos, o herbicida ácido 2,4-diclorofenoxiacético não pode ser considerado dose e tempo-dependentes, mas pode provocar reações alérgicas.

Cancer development and progression vary depending on tumor type, localization, invasion, immunogenicity and the ability of immune system to become activated. There are frequent interactions between tumor cells and immune cells, occuring locally at the site of primary tumor or distally through paracrine signalling of various mediators and cytokines. The main subject of this PhD thesis is to study key factors and aspects of immune response in cancer patients. In the first part, we analyzed immune cells infiltrating tumor tissues of ovarian cancer patients at different stages of disease. We focused on the dynamics of immune response, primarily on frequency of individual T lymphocyte populations in peripheral blood and tumor infiltrating T lymphocytes in tumors of early and advanced stages of ovarian cancer. We found that during disease progression there is a gradual decrease of proinflammatory Th17 and Th1 immune responses and a specific recruitment of regulatory T cells to the tumor site, which results in a significant immune suppression in the tumor microenvironment. In the second part, we demonstrated that the character of immune response in HPV-positive head and neck cancer patients is very different from the patients with tumors not associated with HPV infection. In HPV-positive patients, significantly...

Die heterogene Gruppe der Myopathien kann sowohl die Funktion des Muskels beeinflussen, als auch andere Organsysteme. Erworbenen Muskelerkrankungen sind theoretisch behandelbar, jedoch stehen zumeist nur sehr unspezifische Behandlungsoptionen zur Verfügung, während für vererbte Formen bisher keine kausalen Therapiemöglichkeiten bekannt sind. In dieser Arbeit wurden drei verschiedene Muskelerkrankungen untersucht. Gemeinsam ist ihnen ein jeweils charakteristischer Einstrom von Entzündungszellen, wobei die Zusammensetzung des Zellinfiltrates (z.B. Lymphozyten oder Makrophagen) bei den verschieden Erkrankungen unterschiedlich war. Weiterhin unterscheidet sich das zugrunde liegende Zytokinmilieu für die einzelnen untersuchten Entitäten. Daher war es Ziel der Arbeit, die genauen Interaktionen zwischen den Immunzellen zu untersuchen, sowie die charakteristischen Phänomene der Erkrankungen (Hypoxie, Entzündung und Fibrose). Nekrotisierende Myopathien können sowohl durch eine immun-vermittelte Genese, als auch durch Kontakt mit toxischen Substanzen ausgelöst werden und beide Subgruppen können klar durch morphologische Kriterien, als auch durch spezielle Immunaspekte unterschieden werden. Makrophagen waren hier die vorherrschende Zellpopulation und im gesamten Muskel verteilt. Patienten mit Dermatomyositis dagegen zeigten ein typisches perifaszikuläres Atrophiemuster und hypoxische Effekte, wobei beide Phänomene deutlich ausgeprägter bei juvenilen, als bei adulten Patienten vorkamen. Erbliche Myopathien (z.B. Muskeldystrophie Duchenne) können ebenfalls entzündliche Infiltrate aufweisen und die Entwicklung von Fibrose in der Skelettmuskulatur ist dabei ein Hauptkriterium der Muskelfaserdegeneration. Ein neu entwickelter computer-basierter Algorithmus wurde genutzt, um diese Entwicklung zu quantifizieren. Die Menge an Bindegewebe steigt mit dem Alter der Patienten, während bei älteren Patienten außerdem ein fettgewebiger Umbau ein wichtiger Aspekt der Pathologie war. / The heterogeneous group of myopathies can affect the skeletal muscle or other organ systems and comprise a huge number of different entities. Acquired myopathies are potentially treatable, but there are often only unspecific treatment options, while there is no causative cure for inherited forms of myopathies. In this work, three different entities were analyzed, which all share common aspects of the immune response, but also feature distinct immunological aspects as well. They have an inflammatory part in common, which is mainly regulated by influx of immune cells. However, the composition of these cellular infiltrates (e.g. lymphocytes or macrophages) was varying between the diseases. In addition, the respective cytokine milieu was highly specific in the examined entities. Thus, the aim of the study was to precisely examine interactions between immune cells, and analyze characteristic pathological phenomena (hypoxia, inflammation and fibrosis). Necrotizing myopathies have an immune-mediated background or showed a toxic aetiology and both sub-groups can be distinguished by their morphological characteristics and certain immune aspects. Here macrophages are the predominant cell population and are spread throughout the muscle. Analyses of patients suffering from dermatomyositis showed a typical perifascicular pattern of atrophy, as well as effects of hypoxia and the described features are in general more pronounced in juvenile dermatomyositis than in the adult form. Inherited myopathies (e.g. Duchenne muscular dystrophy) harbor significant inflammatory infiltrates as well and development of fibrosis was a major feature of skeletal muscle degeneration. A computer-based algorithm was used to quantify fibrosis. The amount of connective tissue increased with the age of patients, while at late stage of disease fatty transformation was an additional important issue.

A história natural do câncer cervical invasivo começa com uma infecção produtiva pelo Papiloma vírus humana (HPV) na camada basal do epitélio. Infecções persistentes por vírus de HPVs de alto risco poderão provocar lesões que eventualmente darão origem ao carcinoma invasivo. Sabe-se que o infiltrado inflamatório pode ter um papel importante na evolução da doença. Neste trabalho, quantificamos e caracterizamos fenotipicamente linfócitos T, macrófagos e neutrófilos nas lesões precursoras assim como no carcinoma invasivo. Além disso, para determinar se existia alguma relação entre as populações foi realizada uma análise de correlação entres as populações descritas. Por outro lado, tentando determinar o efeito sistêmico do tumor avaliou-se a frequência de subtipos de monócitos circulantes e através de ensaios alogênicos foi avaliada a capacidade estimuladora das células dendríticas diferenciadas de monócitos circulantes. Os dados apresentados ajudarão a entender o papel que as células do sistema imune podem ter sobre a progressão da doença. / The natural history of cervical cancer begins with a human papilloma virus (HPV) infection of the cells of the basal layer of the epithelium. Persistent infection by high risk HPVs can originate precancerous lesions that may progress to invasive cancer. It has been established the role of the infiltrated inflammatory cells on the progression of the disease. In this work, the phenotype and the frequency of T lymphocytes, macrophages and neutrophils were characterized both in precursor lesions as in invasive carcinoma. In order to stablish a possible relation between the characterized cells, we made a correlation analysis. On the other hand, trying to determine the systemic effect of the tumor we evaluated the frequency of circulating monocyte subtypes and the capacity of dendritic cells to stimulate allogeneic T cells. The data presented will help to understand the role of the immune system cells on the progression of the disease.