Bacterial pathogens cause severe infections worldwide in livestock and in humans, and antibiotic resistance further increases the importance of prophylactic vaccines. Inactivated bacterial vaccines (bacterins) are usually produced via incubation of the pathogen with chemicals such as formaldehyde, which is time consuming and may cause loss of immunogenicity due to the modification of structural components. We evaluated low-energy electron irradiation (LEEI) as an alternative method to generate a bacterin. Rodentibacter pneumotropicus, an invasive Gram-negative murine pathogen, was inactivated with LEEI and formaldehyde. LEEI resulted in high antigen conservation, and LPS activity was significantly better maintained when compared with formaldehyde treatment. Immunization of mice with LEEI-inactivated R. pneumotropicus elicited a strong immune response with no detectable bacterial burden upon sublethal challenge. The results of this study suggest the inactivation of bacteria with LEEI as an alternative, fast and efficient method to generate bacterial vaccines with increased efficacy.

No description available.

Sous irradiation, les flux de particules (neutrons, ions, électrons) créent des défauts ponctuels (lacunes et interstitiels) qui génèrent des défauts étendus (boucles de dislocation, cavités) responsables de la fragilisation des matériaux. L'aluminium est un matériau de faible masse et de faible énergie seuil de déplacement (Ed=16-19 eV) ce qui permet de créer les nanodéfauts étendus directement sous irradiation aux électrons dans un microscope électronique à transmission haute résolution (HRMET). Cet instrument permet non seulement de caractériser finement les nanodéfauts mais également d'étudier leur évolution in situ sous irradiation, ceci en fonction de la température, du taux de dommage et d'une pression partielle d'hydrogène. Nous avons constaté un effet important de la pression partielle d'hydrogène sur la cinétique de formation des boucles de dislocation et des cavités. Parallèlement, nous avons mis en place une modélisation multi-échelle des effets de taille finie et de discrétisation du réseau sur la morphologie d'équilibre des cavités à partir de calculs ab initio des énergies de surface et de liaison de petits amas lacunaires et des amas mixtes lacune-hydrogène. La caractérisation expérimentale des morphologies des cavités en fonction de leur taille met en évidence trois formes régulières : une forme de croix bordée par des surfaces {100}, une forme d'octaèdre non tronqué bordée par des surfaces {111} et une forme d'octaèdre tronqué, bordée par les surfaces {111} et {100} ainsi qu'un effet de taille finie entre ces formes prédit par le modèle. De même, la mesure des fluctuations morphologiques d'une cavité au cours de sa croissance ou de sa décroissance en taille révèle le rôle déterminant des nombres magiques fixés par la géométrie et la discrétisation du réseau. A partir du modèle thermodynamique des cavités et d'une caractérisation expérimentale de la densité volumique et de la distribution en taille des nanodéfauts, nous proposons un modèle de germination des cavités sous irradiation. Nous en déduisons une taille du germe critique et un taux de germination des cavités en fonction de la température et du taux de dommage, qui sont comparés aux valeurs expérimentales correspondantes extraites des observations in situ. Enfin, nous discutons de l'effet de l'hydrogène sur la stabilité des amas mixtes lacune-hydrogène et sur la cinétique de germination des cavités. / Under irradiation, flows of particles (neutrons, ions, electrons) induce ponctual defects (vacancies and interstitials) which generate extended defects (dislocation loops, cavities) that are responsible for materials weakening. Aluminum is a low mass material with a low displacement energy (Ed=16-19 eV) which allows the creation of extended defects directly under electron irradiation in a high resolution transmission electron microscope (HRTEM). This instrument allows not only a fine characterization of nanodefects but also to study their evolution in situ under irradiation, depending on temperature, damage rate and a partial pressure of hydrogen. We noticed an important effect of the hydrogen partial pressure on the kinetic of formation of the cavities and dislocation loops. Simultaneously, we developped a multiscale modelisation of finite size effets and structure discretisation on equilibrium shapes of cavities based on binding and surface energies determined by ab initio calculation of small vacancy clusters and mixted hydrogen vacancy clusters. Experimental characterization of cavity shapes according to the cavity radius reveals three regular shapes : a cross shape bounded by {100} surfaces, an octahedral non-truncated shape bounded by {111} surfaces and an octahedral truncated shape, bounded by {100} and {111} surfaces as well as the determination of a finite size effect predicted by the model. Also, measurements of cavity shape dispersion during its growth and decrease showed a dominant effet of magic numbers, fixed by geometry and frustration of the structure. Based on a thermodynamic model and a fine experimental characterization of volumic density, we propose a nucleation model of cavities under irradiation. We deduce from this model a critical cluster size and a rate of cavity nucleation depending of temperature and damage, which are compared to the corresponding experimental values extracted from in situ observations. Finally, we discuss the effect of hydrogen on the stability of vacancy-hydrogen clusters and on the kinteic of cavity nucleation.

Diese Arbeit liefert die Grundlage für intelligente Beschichtungen aus kovalent gebundenen Polymeren, sogenannten Polymerbürsten, mit Schichtdicken von wenigen Nanometern. Durch ihre Fähigkeit die Oberflächeneigenschaften je nach Umgebungstemperatur, pH Wert und Zusammensetzung zu ändern, kann die Adsorption von Biomolekülen auf eine einzigartige Art und Weise kontrolliert und reguliert werden. Die zusätzliche Modifizierung der Polymerbürsten mit Elektronenstrahlen ermöglicht die Generierung von Gradienten und erweitert die Anwendungsmöglichkeiten in vielfältigen Bereichen der interdisziplinären Forschung, wie z.B. in Hinblick auf das Lab-on-Chip Design oder die Biomedizin. / Polymer brushes represent sensitive coatings with thicknesses of few nanometres and are interesting for interdisciplinary scientific research, e.g. for intelligent lab-on-chip design or biotechnology. It is demonstrated in this work, that polymer brushes are able to change their surface properties depending on the ambient conditions like solvent quality, temperature or pH value. This sensitivity highly influenced the adsorption of bio molecules. Especially mixed polymer brushes enabled the tuning of the adsorbed amount of various proteins and cells. To create surface gradients or structures in the different polymer brush surfaces, controlled electron beam modification was applied.

Thermoplastic Vulcanizates (TPVs) are itself a commercially high valued group of polymer blend. They render technological properties of conventional vulcanized elastomers with the ease of thermoplastic melt (re)processability. With ever growing market, TPVs have got plenty of applications among various fields. Here, the technological properties of these TPVs were tailored according to the purpose by interplaying physical parameters of polymers and advanced high energy electron technology. Electron irradiation, though a well-known technique for cross-linking in polymer industry, is only restricted to final product treatment. We take it to the next level by coupling a conventional internal mixer and a high energy electron accelerator. Polypropylene (PP) and natural rubber (NR) based TPVs have been prepared using this new reactive processing technology, named Electron Induced Reactive Processing (EIReP). Various electron treatment parameters were explored to maximize technological properties of TPVs. Effects of various polyfunctional monomers (PFM) were also studied. In an endeavor to develop a potential method for customization, deep insights of macroscopic and microscopic structure of these TPVs were presented with the help of various advanced scientific characterization techniques. Commonly faced difficulties like viscosity mismatch, cure rate mismatch, and incompatibility due to different molecular structures were furnished along with plausible solutions. Investigation of phase inversion from co-continuous matrix to thermoplastic matrix was dealt with special care as it helps to understand structure property correlation for all TPVs. To make the whole effort relevant, at the end of this thesis a summary of various technological properties has been given for the newly processed and commercially available TPVs.

Diese Arbeit liefert die Grundlage für intelligente Beschichtungen aus kovalent gebundenen Polymeren, sogenannten Polymerbürsten, mit Schichtdicken von wenigen Nanometern. Durch ihre Fähigkeit die Oberflächeneigenschaften je nach Umgebungstemperatur, pH Wert und Zusammensetzung zu ändern, kann die Adsorption von Biomolekülen auf eine einzigartige Art und Weise kontrolliert und reguliert werden. Die zusätzliche Modifizierung der Polymerbürsten mit Elektronenstrahlen ermöglicht die Generierung von Gradienten und erweitert die Anwendungsmöglichkeiten in vielfältigen Bereichen der interdisziplinären Forschung, wie z.B. in Hinblick auf das Lab-on-Chip Design oder die Biomedizin. / Polymer brushes represent sensitive coatings with thicknesses of few nanometres and are interesting for interdisciplinary scientific research, e.g. for intelligent lab-on-chip design or biotechnology. It is demonstrated in this work, that polymer brushes are able to change their surface properties depending on the ambient conditions like solvent quality, temperature or pH value. This sensitivity highly influenced the adsorption of bio molecules. Especially mixed polymer brushes enabled the tuning of the adsorbed amount of various proteins and cells. To create surface gradients or structures in the different polymer brush surfaces, controlled electron beam modification was applied.

Thermoplastic Vulcanizates (TPVs) are itself a commercially high valued group of polymer blend. They render technological properties of conventional vulcanized elastomers with the ease of thermoplastic melt (re)processability. With ever growing market, TPVs have got plenty of applications among various fields. Here, the technological properties of these TPVs were tailored according to the purpose by interplaying physical parameters of polymers and advanced high energy electron technology. Electron irradiation, though a well-known technique for cross-linking in polymer industry, is only restricted to final product treatment. We take it to the next level by coupling a conventional internal mixer and a high energy electron accelerator. Polypropylene (PP) and natural rubber (NR) based TPVs have been prepared using this new reactive processing technology, named Electron Induced Reactive Processing (EIReP). Various electron treatment parameters were explored to maximize technological properties of TPVs. Effects of various polyfunctional monomers (PFM) were also studied. In an endeavor to develop a potential method for customization, deep insights of macroscopic and microscopic structure of these TPVs were presented with the help of various advanced scientific characterization techniques. Commonly faced difficulties like viscosity mismatch, cure rate mismatch, and incompatibility due to different molecular structures were furnished along with plausible solutions. Investigation of phase inversion from co-continuous matrix to thermoplastic matrix was dealt with special care as it helps to understand structure property correlation for all TPVs. To make the whole effort relevant, at the end of this thesis a summary of various technological properties has been given for the newly processed and commercially available TPVs.

Background: With increasing clinical use of NK-92 cells and their CAR-modified derivatives in cancer immunotherapy, there is a growing demand for efficient production processes of these “off-the-shelf” therapeutics. In order to ensure safety and prevent the occurrence of secondary tumors, (CAR-)NK-92 cell proliferation has to be inactivated before transfusion. This is commonly achieved by gamma irradiation. Recently, we showed proof of concept that low energy electron irradiation (LEEI) is a new method for NK-92 inactivation. LEEI has several advantages over gamma irradiation, including a faster reaction time, a more reproducible dose rate and much less requirements on radiation shielding. Here, LEEI was further evaluated as a promising alternative to gamma irradiation yielding cells with highly maintained cytotoxic effector function. Methods: Effectiveness and efficiency of LEEI and gamma irradiation were analyzed using NK-92 and CD123-directed CAR-NK-92 cells. LEE-irradiated cells were extensively characterized and compared to gamma-irradiated cells via flow cytometry, cytotoxicity assays, and comet assays, amongst others. Results: Our results show that both irradiation methods caused a progressive decrease in cell viability and are, therefore, suitable for inhibition of cell proliferation. Notably, the NKmediated specific lysis of tumor cells was maintained at stable levels for three days postirradiation, with a trend towards higher activities after LEEI treatment as compared to gamma irradiation. Both gamma irradiation as well as LEEI led to substantial DNA damage and an accumulation of irradiated cells in the G2/M cell cycle phases. In addition, transcriptomic analysis of irradiated cells revealed approximately 12-fold more differentially expressed genes two hours after gamma irradiation, compared to LEEI. Analysis of surface molecules revealed an irradiation-induced decrease in surface expression of CD56, but no changes in the levels of the activating receptors NKp46, NKG2D, or NKp30. Conclusions: The presented data show that LEEI inactivates (CAR-)NK-92 cells as efficiently as gamma irradiation, but with less impact on the overall gene expression. Due to logistic advantages, LEEI might provide a superior alternative for the manufacture of (CAR-)NK-92 cells for clinical application.