This study examines the role of innate and adaptive immunity in the immunotherapy based on the combination of the ligands stimulating phagocytosis anchored in the tumour cells membrane and the mixture of TLR agonists. This immunotherapy is primarily focused on the innate immunity activation and induces strong inflammatory infiltration, which neutrophils and NK cells are part of. Therefore, the next aim of this study was to evaluate the anti-tumour activity of neutrophils and NK cells. For examination malignant melanoma and pancreatic adenocarcinoma mouse tumour models were used.

The purpose of this study was to assess the impact of virologic monitoring frequencies on treatment failure, adherence to therapy, and the emergence of drug resistance in HIV-1 infected patients. A quantitative, meta-analysis was conducted to investigate the virologic outcomes of infrequent and frequent Viral Load (VL) testing among patient on combination antiretroviral therapy (cART). Data was collected through a self-designed data collection form. Two comparison groups emerged being guided by the VL monitoring frequency. In group I, the health outcomes were compared for (≥3 VLs per year) versus (≤2 VLs per year) and (2 VLs per year) versus (≤1 VLs per year) for group II. Data were analysed using the Cochrane's statistical software, RevMan v5.3. The findings support (2 VLs per year) as the optimal VL monitoring strategy for stable and virologically suppressed patients and there is nothing to be gained by (≥3 VLs per year). / Health Studies / M.P.H.

The aim of this thesis was to find optimal therapy based on combination of membrane-anchored phagocytic ligands (mannose-(G)5-(K)10-STE, mannan-BAM, mannan-SMCC) with LPS (ligand of signal receptor)for treatment of murine melanoma B16-F10. Mixture of mannan-BAM with LPS applied in pulse regime proved to be the most effective, resulting in heigh reduction of tumor growth and significant prolongation of survival.

Nutrientes específicos, denominados farmaconutrientes, demonstraram possuir a capacidade de modular a resposta imunológica e inflamatória de animais e seres humanos, em estudos clínicos e laboratoriais. Dentre os substratos conhecidos, os que têm maior relevância e ação imunomoduladora são a arginina, glutamina, ácido graxo n-3 e nucleotídeos. No entanto, revisões sistemáticas e meta-análises buscam consenso em relação aos vários e controversos resultados publicados sobre os possíveis benefícios da imunonutrição em pacientes críticos. Nossos objetivos foram avaliar a efetividade das dietas enriquecidas com Imunonutrientes na redução de complicações e mortalidade nos diferentes tipos de pacientes críticos. O presente estudo é uma revisão sistemática com metanálise onde foram inseridos ensaios clínicos randomizados avaliando o uso de nutrientes imunomoduladores em doente adulto de ambos os sexos, definido como crítico traumatizado, séptico, queimado ou cirúrgico; as dietas utilizadas deveriam conter um ou mais dos imunonutrientes, em qualquer dose, administradas por via enteral comparadas à dieta padrão pela mesma via em pelo menos um dos grupos de comparação. As bases de dados consultadas foram Pubmed e Cinhal, utilizando os termos: Immunonutrition, arginine, glutamine, n-3, nucleotides e criticall illness. De 206 artigos encontrados inicialmente, apenas 35 preencheram os critérios de elegibilidade estabelecidos. Destes 35 ensaios clínicos, 18 foram conduzidos em pacientes cirúrgicos, 6 em pacientes traumatizados, 5 em pacientes queimados, 5 em pacientes críticos em geral e 1 em pacientes sépticos. Para a população geral, não houve redução significativa do risco de morrer indicada pelo RR de 0,84 (IC de 0,68 a 1,05) e os queimados foram mais protegidos de morrer com RR de 0,25 (IC de 0,09 a 0,66); as complicações infecciosas foram reduzidas com RR de 0,56 (IC de 0,42 a 0,73); a incidência de sepse foi reduzida com RR de 0,45 (IC de 0,29 a 0,69), especialmente nos pacientes traumatizados com RR de 0,42 (IC de 0,26 a 0,68); a incidência de abscesso abdominal também foi reduzida com RR de 0,39 (IC de 0,21 a 0,72) e também de bacteremia com RR de 0,46 (IC de 0,31 a 0,66); o tempo de internação hospitalar diminuiu -3,9 dias (IC de -5,0 a -2,8). Para a população de pacientes cirúrgicos: o óbito não apresentou redução significativa do RR de 0,93 (IC de 0,44 a 1,95), as complicações infecciosas foram reduzidas com RR de 0,51 (IC de 0,41 a 0,63), o tempo de internação hospitalar foi reduzido - 3,41 dias (IC de -4,25 a -2,58) e o tempo de internação em UTI -1,72 dias (IC de -2,12 a -1,31). A utilização de dietas com nutrientes imunomoduladores não alterou a mortalidade em doentes críticos ou cirúrgicos. Indivíduos com mais de 60 anos são menos protegidos de morrer pela utilização de dietas com imunomoduladores. As complicações infecciosas são reduzidas em pacientes críticos com a utilização de imunonutrição, em especial a população de pacientes cirúrgicos. Dietas com mais de 10g/l de arginina protegem mais os pacientes críticos da incidência de complicações infecciosas. Pacientes traumatizados são protegidos da incidência de sepse pela utilização da imunonutrição. O tempo de internação hospitalar e o tempo de internação em UTI foram reduzidos em críticos e cirúrgicos com imunonutrição. A execução de ensaios clínicos explanatórios nos diferentes tipos de doentes críticos com um nutriente imunomodulador isolado, seguida de ensaios pragmáticos de acordo com os resultados dos anteriores, é um fato a ser considerado em futuras pesquisas. / Specific nutrients, the so-called pharmaconutrients, showed a capability to modulate human and animal immune answers in clinical and laboratory studies. Amongst the well-known substracts, arginine, glutamine, fatty accid n-3, and nucleotide showed the most relevant immunomodulator actions. However, systematic revisions and meta-analysis are still in search of agreement due to several controversial results published in the literature on the probable benefits of the immunonutrition of the critically ill. Evaluate the effectiveness of the immunonutrients enriched diets in the reduction of complications and mortality of different types of the critically ill patients. The present study is a meta-analysis systematic revision where randomized clinical trials were included, evaluating the immunomodulator nutrients use in ill adults of both genders, defined as critical traumatized, septicemic, burned, or surgical patients; diets administered should contain one or more immunonutrients, at any dose, ministered by enteral route and compared with a pattern diet offered by the same via at least to one of the comparison groups. Pubmed and Cinhal data banks were searched for the following terms: immunonutrition, arginine, glutamine, n-3, nucleotides and critical illness. In 206 articles were initially found, but only 35 agreed to the established electability criteria: 18 were conducted in the surgical patients, 6 in the traumatized patients, 5 in the burned patients, 5 in the general critical patients, and 1 in the septicemic patients. Considering the general population: obit did not show a significant RR reduction of 0.84 (IC from 0.68 to 1.05); burned patients were more protected from death with RR of 0.25 (IC from 0.09 to 0.66); infections complications were reduced with RR of 0.56 (IC from 0.42 to 0.73); sepsis incidence was reduced with RR of 0.45 (IC from 0.29 to 0.69); specially in the traumatized patients with RR of 0.42 (IC from 0.26 to 0.68); abdominal abscess incidence was also reduced with RR of 0.39 (IC from 0.21 to 0.72) and also bacteremia with RR of 0.46 (IC from 0.31 to 0.66); hospitalization time was also reduced in -3.9 days (IC from -5.0 to -2.8). Considering the surgical patients population: obit did not present a significant RR reduction of 0.93 (IC from 0.44 to 1.95); infections complications were reduced with RR of 0.51 (IC from 0.41 to 0.63); hospitalization period was reduced in -3.41 days (IC from -4.25 to -2.58), and in the ICU time was reduced in -1.72 days (IC from -2.12 to -1.31). Administration of immunomodulator nutrient diets did not change the mortality of the critically ill or surgical patients. Individuals over 60 years are less protected from death with the administration of immunomodulator diets. Infections complications showed a reduction in the critically ill patients with the utilization of immunonutrition, specially the surgical patients population. Diets containing more than 10g/l of arginine protect the critical patients from the incidence of infectious complications. Traumatized patients are protected from the incidence of sepsis by the administration of the immunonutrition. Hospitalization and ICU time were reduced for critical and surgical patients in use of immunonutrition. Explanatory clinical trials in different types of critically ill patients using an isolated immunomodulator nutrient followed by pragmatic trials according to previous results should be considered in future research.

No description available.

The aim of this thesis is to improve the therapeutic effect of the immunotherapy based on the synergy of TLR agonists with phagocytosis stimulating ligands. Furthermore, this thesis is focused on the information transfer to the specific immunity, as well as it pursues the study of the specific immunity relevance during cancer immunotherapy.

The aim of this thesis was to study how to increase effectiveness of cancer immunotherapy based on synergy of compounds stimulating phagocytosis and TLR agonist. Tumor microenvironment was modified by enzymes, which catalised conversion of lactate to pyruvate or acetate. It was monitored effect of enzymes on tumor size, survival of experimental mice and cytotoxicity on tumor cells.

En raison de l'efficacité pour délivrer des antigènes directement dans le cytoplasme des CAPs in vivo, les vecteurs bactériens atténués et basés sur les propriétés du système de sécrétion de type 3 (SST3) attirent de plus en plus l'attention grâce à leur potentiel dans le développement des vaccins contre le cancer. Pseudomonas aeruginosa est un pathogène opportuniste responsable d'infections graves chez les personnes immunodéprimées, les grands brûlés et les patients atteints de la mucoviscidose. Cette pathogénicité repose sur de nombreux facteurs de virulence dont le SST3. Dans nos travaux précédents, le potentiel de souches atténuées de P. aeruginosa dans le domaine de la vaccination anti-tumorale a été démontré. Dans ce travail, nous avons optimisé des vecteurs vaccinaux basés sur le SST3 de P. aeruginosa pour des applications cliniques. Dans un premier temps, la performance de ces vecteurs bactériens a été améliorée en utilisant différents modèles de tumeurs murines. Ceci par : 1) l'ajout d'un épitope spécifique des lymphocytes CD4+ Th aux vecteurs; 2) l'application d'un modèle d'expression bi-antigénique aux vecteurs; 3) la construction de vecteurs induisant une réponse humorale. Dans un deuxième temps, la performance thérapeutique du vecteur bactérien a été optimisée par la modulation de la fréquence des injections et l'intervalle qui les sépare. Cette performance a été confirmée dans des modèles différents de tumeurs murines. Dans un troisième temps, un candidat qui pourrait être appliqué en clinique a été généré par l'adaptation d'un mutant (CHA-OAL) de P. aeruginosa totalement avirulent dans un milieu chimiquement défini. La très faible infectiosité de cette souche a été surmontée par en vaccinant à des emplacements multiples. Par la suite, le potentiel du vecteur bactérien dans l'immunothérapie humaine a été également évalué- dans un premiers temps-dans un modèle de souris humanisées (HHD). Enfin, nous avons observé qu'une immunité anti-vecteur pré-existante n'a pas d'effet sur l'efficacité de la vaccination par le vecteur bactérien. L'ensemble de nos résultats a mis en évidence le potentiel de nos vecteurs vivants et atténués de P. aeruginosa pour des applications dans des essais cliniques pertinents. / Due to the endowed effective ability to deliver antigen to cytoplasm of APCs in vivo, T3SS based attenuated bacterial vectors attracted more and more attention for their potential interest in cancer vaccine development. Pseudomonas aeruginosa est un pathogène opportuniste responsable d'infections graves chez les personnes immunodéprimées, les grands brûlés et les patients atteints de la mucoviscidose. Cette pathogénicité repose sur de nombreux facteurs de virulence dont le système de sécrétion de type III (SSTT). In our previous work, the potential of attenuated P. aeruginosa strains as the carriers for anti-tumor vaccination purpose has been reported. In this work, we would like to strengthen P. aeruginosa T3SS based vaccine vectors and direct the development of these bacterial vectors toward clinical applications. First, the performance of these bacterial vectors has been improved in different murine cancer models by: 1) adding one CD4+ Th epitope to vectors; 2) applying bi-antigen expression pattern to vectors; 3) constructing potential humoral response inducing vectors. Second, the therapeutic performance of bacterial vector has been optimized by modulating injection frequency and interval and then be confirmed in murine tumor models. Third, one clinically applicable candidate has been generated by adapting one totally avirulent P. aeruginosa mutant (CHA-OAL) in a chemically defined medium and the poor infectivity of this new strain has been overcome by vaccinations at multiple loci. Fourth, the potential of bacterial vector for human immunotherapy has been further evaluated in one first level humanized mice (HHD) model. Finally, we observed that the pre-existing anti-vector immunity didn't impair the vaccination efficiency of bacteria vector. Taken together, our results highlight the potentials of our live attenuated P. aeruginosa vectors for applications in relevant clinical trials.

Malgré les programmes de dépistage mis en place et le vaste arsenal thérapeutique disponible, 8,2 millions de décès dans le monde ont été attribués au cancer pour l'année 2012 (données Globocan 2012, OMS). L'immunothérapie antitumorale est en plein essor et consiste notamment à exploiter le système immunitaire de l'hôte pour obtenir une réponse contre la tumeur. L'utilisation de vecteurs bactériens, capables de délivrer un message antigénique et de stimuler de manière concomittante l'immunité innée, fait partie des approches de vaccination antitumorale prometteuses. Parmi ces vecteurs, une bactérie Pseudomonas aeruginosa mise au point par notre laboratoire présente l'intérêt de pouvoir injecter in vivo des antigènes de tumeur, via son système de sécrétion de type III (SST3), directement dans le compartiment intracellulaire des cellules présentatrices d'antigènes. La voie de présentation du CMH I est ainsi favorisée et permet la génération d'une réponse des lymphocytes T cytotoxiques vis-à-vis de la tumeur exprimant l'antigène. Cependant, la poursuite des études précliniques et cliniques paraît délicate, en raison du risque infectieux lié à une bactérie pathogène, quand bien même atténuée. Lors de ce travail, nous avons donc développé une nouvelle souche de P. aeruginosa Killed But Metabolically Active (KBMA), incapable de se répliquer, mais toujours apte à jouer son rôle de vecteur. Une analyse de la réponse immune antitumorale, suite à l'immunisation par différents vecteurs, a permis de mettre en évidence une forte infiltration de la tumeur par des lymphocytes T CD8+ spécifiques de l'antigène, mais également une protection à long terme liée à la présence d'un pool majoritaire de lymphocytes T CD8+ spécifiques effecteurs mémoires. Enfin nous avons cherché à appliquer cette technologie à l'antigène de tumeur anhydrase carbonique 9 (AC9), exprimé par de nombreuses tumeurs solides chez l'homme. / Despite cancer screening programs and the available therapeutic armamentarium, 8.2 million deaths worldwide were due to cancer in 2012 (data Globocan 2012, WHO). The antitumor immunotherapy is booming and aims at using the immune system of the host as a response against the tumor. The use of bacterial vectors, able to deliver an antigenic message and concomitantly stimulate innate immunity, is one of the most promising approaches to antitumor vaccination. Among these vectors, the bacterium Pseudomonas aeruginosa developed by our laboratory has the advantage of being able to inject in vivo tumor antigens via its type III secretion system (T3SS) directly in the intracellular compartment of antigen-presenting cells. The MHC I presentation pathway is thus favored and allows the generation of a cytotoxic T lymphocytes response against antigen-expressing tumors. However, further preclinical and clinical studies remain difficult, because of the risk of infection related to a bacterial pathogen, even if attenuated. In this work, we have developed a new strain of P. aeruginosa Killed But Metabolically Active (KBMA) unable to replicate, but still able to play its role as a vector. An analysis of the antitumor immune response following immunization with different vectors, allowed to demonstrate a strong tumor infiltration by antigen-specific CD8+ T lymphocytes, but also a long-term protection related to the presence of a major pool of antigen-specific effector memory CD8+ T cells. Finally we are seeking to apply this technology to the tumor antigen carbonic anhydrase 9 (CA9), expressed by many solid tumors in humans.

Made available in DSpace on 2015-03-03T14:03:54Z (GMT). No. of bitstreams: 1 Sarah_DVM_DISSERT_PARCIAL.pdf: 746876 bytes, checksum: f638a2adc04ebb6e7a39258725abbfde (MD5) Previous issue date: 2009-09-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Uma glucana insol?vel foi isolada de fermento biol?gico (Saccharomyces cerevisiae), o qual foi submetido a um tratamento com base e o res?duo acidificado. An?lises qu?micas e resson?ncia magn?tica nuclear (NMR) em uma e duas dimens?es (1D e 2D) mostraram que uma ?-(1 3) glucana linear foi purificada, a qual n?o estava contaminada com outros carboidratos, prote?nas ou compostos fen?licos. Os efeitos desta glucana na cicatriza??o de feridas foi avaliado em ?lceras venosas humanas por an?lise histopatol?gica ap?s 30 dias de tratamento. A ?-(1 3) glucana favoreceu a cicatriza??o das ?lceras, promovendo o aumento da hiperplasia epitelial, das c?lulas inflamat?rias, angiog?nese e prolifera??o fibrobl?stica. Este foi o primeiro estudo que investigou o efeito da ?-(1 3) glucana na cicatriza??o de ?lceras venosas em humanos. Os achados sugerem que a glucana ? um potente modificador da resposta biol?gica na cicatriza??o de feridas