Bispyribac-sodium is an acetolactate synthase (ALS) inhibiting herbicide introduced for control of grasses, broadleaves, and sedges in rice and turf. However, Mississippi and Arkansas rice producers have reported inconsistent barnyardgrass (Echinochloa crus-galli Beauv.) control with bispyribac-sodium. Experiments were conducted to determine if inconsistent barnyardgrass control with bispyribac-sodium could be alleviated through adjuvant technology. Absorption of 14C-bispyribac-sodium was greatest with a proprietary blend of methylated seed oil/organosilicone (MSO/OSL) (0.37 L ha-1) plus urea ammonium nitrate (UAN) (2% v/v) and the proprietary blend of MSO/OSL/UAN (2% v/v) at 80 and 74% of applied 14C-bispyribac-sodium, respectively. The addition of 32% UAN to MSO/OSL and non-ionic organosilicone (OSL/NIS) adjuvant systems resulted in a four- to fiveold increase in absorption compared to treatments without UAN. Maximum absorption was achieved 12 h after application and therefore, bispyribac-sodium should be rainfast at this time. MSO/OSL adjuvants supplied from multiple manufacturers were also examined to determine which provided the highest levels of 14C-bispyribac-sodium absorption as well as herbicidal efficacy and rainfastness. The addition of Rivet® plus UAN or Sil-MES 100® plus UAN to 14C-bispyribac-sodium resulted in the highest levels of absorption among adjuvants supplied. Bispyribac-sodium applied with no adjuvant or with 32% UAN only resulted in significant reductions in control compared to MSO/OSL adjuvant formulations. Addition of MSO or MSO/OSL adjuvants and UAN to bispyribac-sodium resulted in greater than 85% freshweight reduction of barnyardgrass. Bispyribac-sodium applied with MSO or Dyne-A-Pak® was rainfast by two and four hours after application, respectively. Addition of Sil-MES 100® plus UAN or Inergy® plus UAN to bispyribac-sodium reduced the rainfastness interval to 30 minutes and one hour after herbicide application, respectively. Further experiments were conducted to determine if individual components of 32% UAN as well as ammonium sulfate (AMS) would provide control similar to that of 32% UAN. The addition of ammonium chloride, calcium nitrate, and ammonium sulfate at selected rates provided barnyardgrass control similar to that of 32% UAN.

Today's development of novel vaccines stresses the need for edible vaccines that are inexpensive, easily administered and capable of being stored and transported without refrigeration. Without these characteristics, developing countries find it difficult to adopt vaccination as the central strategy for preventing their most devastating diseases. A promising approach is the production of vaccines in plants we commonly consume. Two major obstacles have been encountered in developing vaccines in plants. First, the expression level of foreign antigens tends to be low and second, co-expression of an adjuvant may be required to facilitate an appropriate immune response. Ricin, a plant toxin that survives the human digestive process, has been proven to stimulate an immune response and could therefore serve as a suitable adjuvant. The long-term goal is to produce a vaccine that protects against the disease entamoebic dysentery. The specific goal of this research was to produce ricin in tobacco as adjuvant for the vaccine. Vectors were constructed that fused the ricin coding sequence to different plant promoters and transgenic tobacco plants were generated by transformation with Agrobacterium tumefaciens. The levels of expression in these transgenic plants were tested using immunoblot assays. Southern blot analysis was performed for the highest expressors of each construct. The enzymatic activity of the tobacco-synthesized ricin was shown using a protein translation inhibition assay. Expression of ricin was also confirmed using transient transformation of hairy root cultures. Future experiments will address the practical use of the tobacco-synthesized ricin as adjuvant, as well as the expression of the ricin B subunit fused to a protective antigen of Entamoeba histolytica in tobacco as edible vaccine. / Master of Science

Les vecteurs nanoparticulaires comme systèmes de relargage contrôlé pour des applications vaccinales font l’objet d’intenses recherches, notamment dans le domaine du VIH1. Une approche novatrice consiste à co-administrer des molécules immuno-stimulatrices avec les antigènes d’intérêt, afin d’amplifier le recrutement et l’activation des cellules dendritiques (DCs). Un tel vecteur vaccinal stimulerait l’intensité de la réponse immunitaire et une immunité au niveau des muqueuses vaginales et anales pourrait être obtenue après vaccination. Des nanoparticules de poly(acide lactique) (NP-PLA) ou de chitosane/sulfate de dextrane (NP-CSD) ont été utilisées comme véhicules et adjuvants de protéines du VIH1, gp140 et p24. Le poly(I:C), ligand de TLR3 est la molécule immuno-stimulatrice retenue pour ses propriétés adjuvantes. Les NP-PLA et NP-CSD présentent un potentiel équivalent pour l’adsorption de protéines. Par contre, si les NP-CSD permettent l'adsorption du poly(I:C) (95%), elle est moins reproductible sur les NP-PLA. Pour chaque formulation, la capacité à induire in vitro la maturation des DCs a été évaluée en suivant les marqueurs CD25, CD80, CD83, par cytométrie en flux. L’adsorption de poly(I:C) sur les NP-PLA ou les NP-CSD amplifie les capacités de maturation de ces nanoparticules, un effet synergique étant observé avec les NP-CSD. Nos travaux montrent que la co-adsorption d’un TLR ligand, avec des antigènes protéiques du VIH sur des nanoparticules biodégradables, est possible et confère à la formulation vaccinale un effet immuno-stimulant in vitro. In vivo, les formulations vaccinales contenant du poly(I:C) induisent de très forts taux d’anticorps sériques chez la souris. / Use of nanoparticulate vectors in vaccination as controlled release systems based on biodegradable polymers has been widely studied, particularly for HIV vaccine research. An innovative approach is to co-administer antigens of interest with immuno-stimulatory molecules to amplify the recruitment and activation of dendritic cells (DCs). Such a vaccine candidate could boost the intensity of the immune response, and mucosal immunity in vaginal and anal secretions could be obtained after vaccination.We used nanoparticles of poly(lactic acid) (NP-PLA) or chitosan / dextran sulfate (NP-CSD), as vehicles and adjuvants for HIV-1 proteins, gp140 and p24. Poly (I:C), TLR3 ligand molecule, is the immuno-stimulatory molecule chosen for its adjuvant properties. The NP-PLA and NP-CSD have shown their great potential as carriers of proteins. By cons, if NP-CSD allows the adsorption of poly(I:C) with a yield of 95%, the adsorption is less reproducible on NP-PLA. For each formulation, the ability to induce in vitro maturation of DCs was evaluated by following the marker CD25, CD80, CD83, by flow cytometry. Adsorption of poly(I:C) on the NP-PLA or the NP-CSD amplifies the maturation abilities of particles and has a synergistic effect with the NP-CSD.Our work shows that co-adsorption of a TLR ligand with HIV protein antigens onto biodegradable nanoparticles is possible and gives an immuno-stimulant effect to the vaccine formulation in vitro. In vivo, vaccine formulations containing poly(I:C) induce very high levels of serum antibodies in mice.

Featured adjuvant chemotherapy treatment in women with breast cancer can lead to pain. The aim of this study was to explore, the variety of perceptions and impact of adjuvantchemotherapy-induced pain in daily life, of some women newly diagnosed with breast cancer.Inclusion criteria were participating in an ongoing stress management projectand chemotherapy of (anthracycline/taxan) in doses of 75mg² or more. Exclusioncriteria were inability to understand and communicate in Swedish and mentalillness. After ethical approval of the sub study in September 2010, women wereconsecutively included through oral and written request. Phenomenologicalapproach was used in the eight interviews and data analysis. The resultconsisted of five categories of description, the obvious pain, themanageable pain, the lonely pain, the unimaginable pain andultimately the crippling pain. The existence was open when the pain feltdescribable and manageable, while it was concluded when the pain seemedinexplicable and life drastically changed. The study showed a significantpainful impact of chemotherapy. The woman had difficulties to refer to theinformation given by the medical services, when the pain went beyond previousexperiences. There was a tendency that the woman waited before she contactedthe medical services, this waiting made room for difficult thoughts andfeelings. A question for further research is how the staff can capture andbetter help the women who experience severe pain. / Dagens adjuvanta cytostatikabehandlingav kvinnor med bröstcancer kan leda till smärta. Denna studies syfte var att undersöka olikauppfattningar om inverkan av smärta, utlöst av adjuvant cytostatikabehandling,på dagligt liv hos några kvinnor som nyligen diagnostiserats med bröstcancer.Inklusionskriterier var deltagande i ett pågående stresshanteringsprojekt ochcytostatikabehandling i doser om 75mg² eller mer av antracyklin och/ellertaxan. Exklusionskriterier var oförmåga att förstå och kommunicera på svenska ochpsykisk sjukdom. Efter etiskt godkännande av delstudien i september 2010 inkluderades kvinnorna konsekutivt genommuntlig och skriftlig förfrågan. Fenomenologisk ansats användes i de åttaintervjuerna och i resultatbearbetningen. Resultatet kom att utgöras av fembeskrivningskategorier: den förklarliga smärtan, den övervinneliga smärtan, denensamma smärtan, den ofattbara smärtan och sist den förlamande smärtan.Tillvaron var öppen då smärtankändes förklarbar och därmed hanterbar, medan den slöts när smärtan kändesoförklarlig och kvinnornas liv förändrades drastiskt. Studien visar på en betydande smärtinverkan vid cytostatikabehandling. Kvinnornafick svårt att referera till den av sjukvården givna informationen, när smärtangick utanför tidigare beskrivning och smärtupplevelser. Det fanns en tendensatt kvinnan avvaktade innan hon kontaktade sjukvården och i denna väntanuppstod svåra tankar och känslor. En fråga för vidare forskning är hurpersonalen kan fånga upp och bättre hjälpa de kvinnor som får svåra smärtor.

De nombreux obstacles freinent le développement d'un vaccin efficace contre le VIH. Pour franchir ces barrières, le recours aux adjuvants est une option prometteuse. Dans ce contexte, l'objectif de ce doctorat est l'évaluation de combinaisons de ligands de PRR et de particules biodégradables pour la vaccination muqueuse. La première partie de l'étude vise à analyser la valeur ajoutée de molécules hybrides composée de deux ligands de PRR comparée à la co-administration des deux agonistes. Des molécules stimulant TLR7 et TLR2 puis TLR7 et NOD2 ont été évaluées. Nous avons démontré l'intérêt de l'association de ligands de PRR au sein d'une même molécule, pour l'induction de réponses immunitaires systémiques et muqueuses. Des études récentes ont montré l'intérêt d'agonistes de STING comme adjuvant vaccinaux. Nous avons étudié l'induction de réponses immunitaires par les agonistes de STING, administrés par voie parentérale ou muqueuse. Nous avons démontré le fort potentiel de ligands de STING pour l'induction de réponses cellulaires et muqueuses. Lors de ces études, nous avons démontré que l'intérêt de la vectorisation d'agonistes de PRR dépend de la molécule. En effet, bien que la vectorisation d'une molécule hybride TLR7/TLR2 n'ait pas d'impact sur la réponse immunitaire induite, la vectorisation d'agonistes de STING potentialise leur effet immunostimulant. Pour finir, nous avons montré que la voie d'administration a un impact sur la réponse immunitaire induite. Afin de mieux comprendre les mécanismes mis en jeu, une étude de biodistribution des formulations de NP de PLA après administration par voie systémique ou muqueuse a été réalisée / There are many barriers to the development an effective HIV vaccine. The use of adjuvants is a promising option to overcome these obstacles. In this context, the objective of this PhD is the evaluation of combinations of PRR ligands and biodegradable particles for mucosal vaccination.The first part of this study aimed at assessing the added value of hybrid molecules composed of two PRR ligands compared to the co-administration of the two agonists. TLR7 and TLR2 stimulating molecules followed by TLR7 and NOD2 were evaluated. We demonstrated the interest of the association of PRR ligands within the same molecule for the induction of systemic and mucosal immune responses.Recent studies showed the interest of STING agonists as a vaccine adjuvant. We investigated the induction of immune responses by STING agonists administered parenterally or mucosally. We confirmed the strong potential of STING ligands for the induction of cellular and mucosal responses.In these studies, we demonstrated that the interest of vectorization of PRR agonists depends on the molecule. Indeed, although the encapsulation of a TLR7/TLR2 hybrid molecule has no impact on the induced immune response, the vectorization of STING agonists potentiates their immunostimulatory effect.Finally, we showed that the route of administration has an impact on the immune response induced. In order to better understand the mechanisms involved, a biodistribution study of PLA NP formulations after systemic or mucosal administration was performed

nÃo hà / O cÃncer de mama foi descrito hà muitos anos e documentado, pela primeira vez, por Imhotep, mÃdico, astrÃlogo e arquiteto egÃpcio, nascido em 2.650 antes de Cristo (a.C.) que recomendava Ãquela Ãpoca, como tratamento, a cauterizaÃÃo do tecido doente. Tamoxifeno à o fÃrmaco mais prescrito no tratamento do cÃncer de mama. Sua utilizaÃÃo à principalmente na modalidade adjuvante, em pacientes prà ou pÃs menopausadas, receptor de estrÃgeno e/ou progesterona positivos. à tambÃm utilizado no tratamento da doenÃa localmente avanÃada e metastÃtica e em menor proporÃÃo nas pacientes com contra-indicaÃÃo formal de cirurgia ou que se recusam a se submeter a esta modalidade de tratamento. Na neo-adjuvÃncia à utilizada apenas em ensaios clÃnicos. O tamoxifeno tambÃm diminui, na adjuvÃncia por cinco anos, a probabilidade de recidiva em 47% e de morte por cÃncer de mama em 26% e os dois principais efeitos colaterais, apesar de raros, sÃo aumento da prevalÃncia de cÃncer de endomÃtrio e de fenÃmenos tromboembÃlicos. Este estudo teve como objetivo principal avaliar as pacientes portadoras de cÃncer de mama, no Instituto do CÃncer do CearÃ, tratadas com tamoxifeno de forma adjuvante, no perÃodo de janeiro de 1993 a 1996, com relaÃÃo aos principais benefÃcios e efeitos colaterais, bem como anÃlise de sobrevivÃncia. ProntuÃrios de setecentos e quarenta e duas pacientes foram analisados no que diz respeito aos dados sÃcio- demogrÃficos, idade, status menopausal, estadiamento clÃnico e patolÃgico, dosagem de receptores de estrÃgeno e progesterona, casos de cÃncer de endomÃtrio, principais sÃtios de metÃstases, modalidade de tratamento cirÃrgico, radioterÃpico e quimioterÃpico, causas de Ãbito, tipo histolÃgico, status dos linfonodos axilares e anÃlise de sobrevivÃncia de acordo com o estadiamento. Concluiu-se que a maioria dos dados estÃo de acordo com a literatura e que o prejuÃzo da anÃlise foi resultante da qualidade dos registros realizados nos prontuÃrios, devendo cada vez mais os mÃdicos serem estimulados a documentar, de forma clara e legÃvel, o maior nÃmero de informaÃÃes possÃveis, nÃo apenas as positivas, mas todas aquelas que, mais freqÃentemente, possam ter relaÃÃo com a utilizaÃÃo de qualquer medicamento prescrito. / O cÃncer de mama foi descrito hà muitos anos e documentado, pela primeira vez, por Imhotep, mÃdico, astrÃlogo e arquiteto egÃpcio, nascido em 2.650 antes de Cristo (a.C.) que recomendava Ãquela Ãpoca, como tratamento, a cauterizaÃÃo do tecido doente. Tamoxifeno à o fÃrmaco mais prescrito no tratamento do cÃncer de mama. Sua utilizaÃÃo à principalmente na modalidade adjuvante, em pacientes prà ou pÃs menopausadas, receptor de estrÃgeno e/ou progesterona positivos. à tambÃm utilizado no tratamento da doenÃa localmente avanÃada e metastÃtica e em menor proporÃÃo nas pacientes com contra-indicaÃÃo formal de cirurgia ou que se recusam a se submeter a esta modalidade de tratamento. Na neo-adjuvÃncia à utilizada apenas em ensaios clÃnicos. O tamoxifeno tambÃm diminui, na adjuvÃncia por cinco anos, a probabilidade de recidiva em 47% e de morte por cÃncer de mama em 26% e os dois principais efeitos colaterais, apesar de raros, sÃo aumento da prevalÃncia de cÃncer de endomÃtrio e de fenÃmenos tromboembÃlicos. Este estudo teve como objetivo principal avaliar as pacientes portadoras de cÃncer de mama, no Instituto do CÃncer do CearÃ, tratadas com tamoxifeno de forma adjuvante, no perÃodo de janeiro de 1993 a 1996, com relaÃÃo aos principais benefÃcios e efeitos colaterais, bem como anÃlise de sobrevivÃncia. ProntuÃrios de setecentos e quarenta e duas pacientes foram analisados no que diz respeito aos dados sÃcio- demogrÃficos, idade, status menopausal, estadiamento clÃnico e patolÃgico, dosagem de receptores de estrÃgeno e progesterona, casos de cÃncer de endomÃtrio, principais sÃtios de metÃstases, modalidade de tratamento cirÃrgico, radioterÃpico e quimioterÃpico, causas de Ãbito, tipo histolÃgico, status dos linfonodos axilares e anÃlise de sobrevivÃncia de acordo com o estadiamento. Concluiu-se que a maioria dos dados estÃo de acordo com a literatura e que o prejuÃzo da anÃlise foi resultante da qualidade dos registros realizados nos prontuÃrios, devendo cada vez mais os mÃdicos serem estimulados a documentar, de forma clara e legÃvel, o maior nÃmero de informaÃÃes possÃveis, nÃo apenas as positivas, mas todas aquelas que, mais freqÃentemente, possam ter relaÃÃo com a utilizaÃÃo de qualquer medicamento prescrito. / Breast cancer is a disease that was described many years ago and has been documented, for the first time, by Imhotep, physician, astrologer and Egyptian architect, born in 2.650 before Christ (b.C.), who recommended, at that time, as a way of treatment, cauterization of the diseased tissue. Tamoxifen is the drug more prescribed in the treatment of breast cancer. Itâs use is mainly in the adjuvant modality, in pre or post menopaused patients positive estrogen and/or progesteron receptors. Itâs used in the treatment of locally advanced and metastatic disease and in smaller proportion in patients with formal contraindication of surgery or that refuse to submit this treatment modality. In the neoadjuvancy it is just used in clinical research. The tamoxifen also reduces in the adjuvant modality during five years, the probability of recurrence in 47% and deaths caused by breast cancer in 26% and the two main side effects, in spite of rare, are the increase of the prevalence of endometrial cancer and of thromboembolic phenomenas. This study had as main objective to evaluate the patients, breast cancer bearers, in the Institute of Cancer of CearÃ, treated with tamoxifen in the adjuvant form in the period of 1993 to 1996 regarding the main benefits and side effects, as well as survival analysis. Seven hundred forty-two patientsâprontuaries were analyzed in respect to the demographic datas, age, menopausal status, clinical and pathological staging, dosage of estrogen and/or progesterone receptors, cases of endometrial cancer, main local metastasis, modality of surgical treatment, radiotherapy and chemotherapy, death causes, histological type, status of the axillary lymph nodes and survival analysis in agreement with the staging. We concluded that most of the data is in agreement with the literature and that the demage of the analysis was resulting from the quality accomplished found in the prontuaries. Also, doctors should be more and more stimulated to document, in a clear and readable way, the largest number of possible information, not just the positive ones, but all those that more frequently can have relationships with the use of any prescribed medicine. / Breast cancer is a disease that was described many years ago and has been documented, for the first time, by Imhotep, physician, astrologer and Egyptian architect, born in 2.650 before Christ (b.C.), who recommended, at that time, as a way of treatment, cauterization of the diseased tissue. Tamoxifen is the drug more prescribed in the treatment of breast cancer. Itâs use is mainly in the adjuvant modality, in pre or post menopaused patients positive estrogen and/or progesteron receptors. Itâs used in the treatment of locally advanced and metastatic disease and in smaller proportion in patients with formal contraindication of surgery or that refuse to submit this treatment modality. In the neoadjuvancy it is just used in clinical research. The tamoxifen also reduces in the adjuvant modality during five years, the probability of recurrence in 47% and deaths caused by breast cancer in 26% and the two main side effects, in spite of rare, are the increase of the prevalence of endometrial cancer and of thromboembolic phenomenas. This study had as main objective to evaluate the patients, breast cancer bearers, in the Institute of Cancer of CearÃ, treated with tamoxifen in the adjuvant form in the period of 1993 to 1996 regarding the main benefits and side effects, as well as survival analysis. Seven hundred forty-two patientsâprontuaries were analyzed in respect to the demographic datas, age, menopausal status, clinical and pathological staging, dosage of estrogen and/or progesterone receptors, cases of endometrial cancer, main local metastasis, modality of surgical treatment, radiotherapy and chemotherapy, death causes, histological type, status of the axillary lymph nodes and survival analysis in agreement with the staging. We concluded that most of the data is in agreement with the literature and that the demage of the analysis was resulting from the quality accomplished found in the prontuaries. Also, doctors should be more and more stimulated to document, in a clear and readable way, the largest number of possible information, not just the positive ones, but all those that more frequently can have relationships with the use of any prescribed medicine.

En 2014 la Tuberculose (TB) à dépassé le VIH comme la principale cause de décès par maladie infectieuse dans le monde soulignant le besoin urgent de développer un vaccin plus efficace contre cette maladie. Le candidat vaccin contre la TB, ID93/GLA-SE, dévéloppé à l’Infectious Disease Research Institute (IDRI), est aujourd’hui en essai clinique de phase IIa et a montré des résultats pré-cliniques et cliniques promettants. Dans de modèle murin de TB, ce vaccin induit une forte réponse TH1, considérée comme centrale dans la protection contre la TB, et la production d’IgG2 par les lymphocytes B. Néanmoins, les mécanismes d’action de GLA-SE sont encore peu connus.L’objectif principal de cette thèse est donc d’élucider les méchanismes clés qui relient les réponses innées et adaptatives induites par cet adjuvant dans le modèle murin. Un objectif secondaire est d’établir un modèle murin de rechute de TB après traitement et d’évaluer l’utilisation d’ID93/GLA-SE en tant que vaccin immuno-thérapeutique et sa capacité à réduire les taux de rechute dans ce modèle. L‘ensemble de ce travail nous a permis de mieux comprendre les mécanismes impliqués dans la réponse immunitaire adaptative induite par GLA- SE et de montrer la capacité de ID93/GLA- SE a être utilisé comme un vaccin thérapeutique contre la tuberculose dans le but de réduire les taux de rechute post-thérapeutiques. / In 2014 tuberculosis (TB) surpassed HIV as the leading cause of death by an infectious disease worldwide emphasizing the urgent need to develop a more effective vaccine against this airborne disease. The Infectious Disease Research Institute (IDRI) TB candidate vaccine ID93/GLA-SE is currently undergoing a Phase IIa clinical trial and has shown promising preclinical and clinical results. In murine models of TB this vaccine drives a strong CD4 TH1 response, which is thought to be important for protection against TB, and an IgG2c skewed B cell response. However, little is known about the cellular and molecular events that drive GLA-SE adjuvanticity.To that end, the main objective of my thesis was to elucidate the key mechanisms that connect innate and adaptive immune responses elicited by this adjuvant in the murine model. A secondary objective was to evaluate the possibility of using ID93/GLA-SE as adjunct therapy to existing antibiotic treatments to reduce relapse rates after TB treatment.Collectively the results obtained during this research project and thesis broaden our knowledge and our current understanding of the mechanisms involved in the adaptive immune response induced by GLA-SE and show the capacity of ID93/GLA-SE to be used as a therapeutic vaccine against TB to reduce post-therapeutic relapse rates.

Clostridium difficile est une bactérie anaérobie stricte responsable de diarrhées consécutives à une antibiothérapie et de colites pseudomembraneuses. La destruction du microbiote intestinal de barrière favorise l’implantation de C. difficile, qui se multiplie, adhère aux cellules épithéliales et produit les toxines TcdA et TcdB. Les infections à C. difficile sont devenues un problème majeur de santé publique. En particulier les nombreuses rechutes posent un problème thérapeutique. L’immunisation active est une des approches permettant de prévenir les rechutes et réduire l’incidence des infections. Plusieurs essais de vaccination ciblant les toxines sont en cours de développement mais sont sans action sur la première étape de l'infection à C. difficile, la colonisation intestinale. Notre objectif a été de développer une stratégie vaccinale par voie muqueuse pour lutter contre la colonisation intestinale. Des protéines de surface de C. difficile ainsi que la toxine TcdB ayant un rôle dans la pathogénicité ont été sélectionnées et utilisées comme cibles vaccinales. Certains candidats vaccinaux ont été encapsulés afin de pouvoir délivrer les antigènes au niveau de la muqueuse intestinale par voie orale. De plus, pour induire une réponse immunitaire localisée au niveau de la muqueuse intestinale, différents adjuvants ont été testés. Nous avons analysé après vaccination la réponse immunitaire induite au niveau local et systémique. Pour identifier de nouveaux candidats vaccin, le rôle de différentes protéines de surface dans la colonisation intestinale de C. difficile a été caractérisé. L'ensemble des essais in vivo a été mené dans deux modèles animaux de référence pour les infections à C. difficile : la souris et le hamster, pour suivre respectivement la colonisation intestinale par C. difficile et le taux de survie après infection. / Clostridium difficile is an anaerobic bacterium that is responsible for post-antibiotic diarrhea and pseudomembranous colitis. The disruption of the intestinal microbiota barrier effect promotes the establishment of C. difficile, which multiplies, adheres to epithelial cells, and produces the toxins TcdA and TcdB. C. difficile infections are considered as a major public threat. In particular, the multiple recurrences are difficult to treat. Active immunization is one of the new approaches to reduce recurrences and the incidence of these infections. Several vaccine targeting toxins are in development. However, these vaccines have no effect on the first step of C. difficile infection, the host colonization. The objective was to develop a mucosal vaccine strategy to act on intestinal colonization. Surface proteins of C. difficile and TcdB involved in the pathogenicity process were selected and used as vaccine targets. These antigens were produced and encapsulated to deliver them to the intestinal mucosa. Furthermore, to induce a gut mucosal immunity, different adjuvants were tested. After vaccination, we analyzed the local and systemic immune response by immunoassays. Finally, in order to characterize new vaccine candidates, the role of different surface proteins in C. difficile colonization was evaluated. These vaccine trials were conducted in two animal models of reference for C. difficile infections: the mouse and the hamster models, which permit to follow the colonization and the survival rate after infection, respectively.

La sarcoïdose est une maladie inflammatoire multisystémique s'attaquant aux poumons dont l'étiologie demeure inconnue. Elle est caractérisée par l'accumulation de granulomes noncaséeux dans les organes touchés. L'appareil pulmonaire est fréquemment atteint par la sarcoïdose et d'importants changements de population cellulaire sont observés dans le fluide récolté lors de lavages bronchoalvéolaires (LBA). Une étude précédente a démontré la possibilité d'utiliser un modèle animal de rat pour l'étude de cette maladie. Cette étude a démontré que l'adjuvant complet de Freund (CFA) administré par voie intraveineuse (IV) à deux reprises induit l'apparition de granulomes qui furent, par la suite, analysés par tomographie axiale. Le projet de recherche décrit dans ce mémoire a visé à développer, à partir du modèle déjà connu, un modèle mieux toléré par les animaux et à analyser les caractéristiques cellulaires, histologiques et immunologiques de celui-ci. De plus, certains agonistes des récepteurs nicotiniques, molécules ayant des propriétés anti-inflammatoires, seront testés sur le modèle comme traitement potentiel. Il fut démontré lors de cette étude qu'une seule injection de CFA cause l'apparition de granulomes dans les poumons des animaux et que cette injection est bien tolérée par les animaux. Les analyses cellulaires des LBA des animaux malades ont permis de noter une forte similarité avec la sarcoïdose pulmonaire humaine. De plus, le profil immunologique des animaux était très près de la pathologie humaine. Finalement, l'utilisation des agonistes nicotiniques n'a pas permis d'atténuer l'inflammation reliée à cette pathologie granulomateuse.

No description available.