Helmintheninfektionen induzieren in ihren Wirten Typ 2 (Th2) Immunantworten, welche parasitäre Larvenstadien hemmen und/oder zur Abstoßung von intestinalen Würmern führen. Th2-Antworten können auch gegenüber Umweltallergenen ausgebildet werden und allergische Reaktionen vermitteln. Mastzellen (MZ) spielen eine herausragende Rolle für die Wirtsantwort gegen Parasiten. Die Ergebnisse der vorliegenden Arbeit zeigen, dass eine Infektion von MZ-defizienten Mäusen mit Helminthen zu einer reduzierten Produktion von IL-25, IL-33 und TSLP, einer beeinträchtigten Etablierung von Th2 Immunantworten und einer erhöhten Wurmlast führt. Diese Parameter konnten allerdings nach einem erfolgreichen Transfer von Knochenmark aus Wildtypmäusen wiederhergestellt werden. Die vorliegende Arbeit beschreibt somit eine wichtige Funktion von Mastzellen für die Initialisierung von Th2 Immunantworten. Des Weiteren konnte gezeigt werden, dass ein Cystein-Protease-Inhibitor - AvCystatin sowohl Parameter von Atemwegsentzündung und -hyperreaktivität als auch Lieschgraspollen (Phl p 5b)-spezifische Immunantworten in einem klinisch relevanten Mausmodell für Atemwegsentzündung inhibiert. Gleichzeitig führte die Behandlung mit AvCystatin zu einem Anstieg von IL-10 und erhöhten Frequenzen von CD4+CD25+Foxp3+ T Zellen. Die in vivo Effekte von AvCystatin wurden unabhängig von der Protease-inhibitorischen Aktivität des Immunmodulators oder dessen Fähigkeit zur Oligomerisation vermittelt. AvCystatin unterdrückte zudem die Allergen-spezifische Produktion von IL-13 und induzierte in vitro unter Verwendung mononukleärer Zellen aus dem peripheren Blut (PBMCs) von Graspollen allergischen Patienten einen IFN-gamma vermittelten Th1 shift. Zusammenfassend tragen die Ergebnisse der Arbeit zu einem besseren Verständnis für die frühen Ereignisse von Th2 Immunantworten bei und zeigen, dass Helminthenmoleküle Bystandereffekte auf Immunantworten vermitteln können, die gegen andere Antigene gerichtet sind. / Helminth infections induce protective type 2 (Th2) immune responses in the host leading to arrested larval development and/or intestinal worm expulsion. Moreover, Th2 immune responses are initiated against harmless environmental allergens and mediate development of allergic disease. Among multiple mechanisms implicated in host responses to parasites and allergens, mast cells (MCs) play a pivotal role. The present study shows that MC-deficient mouse strains following infection with two gastrointestinal helminths had dramatically reduced early production of the tissue-derived cytokines IL-25, IL-33, and TSLP, which resulted in impaired induction of Th2 immune responses as well as increased parasite burden. These parameters were restored after transfer of WT bone marrow. These data reveal an important role for MCs in orchestrating type 2 immune responses. Parasites have developed various strategies to modulate the immune system via induction of a range of regulatory mechanisms. In this study AvCystatin, the filarial cysteine inhibitor, was found to inhibit airway inflammation and hyperreactivity induced by a clinically relevant allergen of timothy grass pollen (Phl p 5b). AvCystatin increased levels of the regulatory cytokine IL-10 and total numbers of CD4+CD25+Foxp3+ T cells. The immunomodulatory effect in vivo was found to be independent of AvCystatin’s protease inhibitor activity or oligomerization. Finally, AvCystatin suppressed allergen-specific production of IL-13 and created a shift towards Th1 immunity by increased levels IFN-gamma of human peripheral blood mononuclear cells (PBMCs) from grass pollen allergic patients. The findings contribute to a better understanding of the early events that dictate the priming of type 2 immune responses. Furthermore, helminth product-induced suppression may also have effects on bystander responses to unrelated antigens, thus, suggesting a promising preventive and therapeutic concept in the treatment of aberrant inflammations.

<p>Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. </p><p><i>In vitro</i> tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary.</p><p>Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD).</p><p>In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (<i>Phleum pratense</i>) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD.</p><p>Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp <i>Penaeus aztecus</i> (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach.</p><p>In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant protein, to be used as a spiking reagent.</p>

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant protein, to be used as a spiking reagent.

MRO-industrin är en kritisk del för underhåll inom flygindustrin och arbetsprocesserna ser olika ut från företag till företag. För att kunna utföra ett så säkert och kvalitetsmässigt arbete som möjligt så är det viktigt att riskbedöma samt minimera existerande risker i operationen för att säkerställa kvalitet till kund samt säkerhet för arbetare. Uppdragsgivaren ST Engineering Aerospace Solutions (STEAS) är det ledande företaget i Sverige när det gäller MRO arbete och med den ständiga utvecklingen av utrustning upptäcks det allt fler risker som kräver analyser och minimering. Forskning har tidigare gjorts på hur fixtur design kan förbättras ergonomiskt m h a virtuell miljöteknik. Detta för att kunna minimera design och tillverkningskostnader med hänsyn till itereringsprocessen. Detta i kombination med studier gjorda på muskuloskeletala skador med kopplingar till MRO-industrin belyser behovet av riskanalys i MRO industrin. Arbetet hos STEAS går ut på att utföra riskanalyser ur ett tekniskt, ergonomiskt och ekonomiskt perspektiv på deras rörliga fixturer samt kemiska processer som behandlar diverse komponenter av motornacellet. Detta för att STEAS sedan ska kunna kolla på eventuella kostnader och implementera de rekommenderade åtgärderna. Analysmetoderna som används är en PHL (Preliminary Hazard List), FMEA (Failure Modes and Effects Analysis) och en ACHB analys (Accidental Cost &amp; Hypothetical Benefit) / The MRO industry is a critical part of maintenance in the aviation industry where the work processes differ from company to company. In order to be able to perform as safe and high-quality work as possible, it is important to assess risks and minimize existing risks in the operation to ensure quality for customers and safety for workers. The client ST Engineering Aerospace Solutions (STEAS) is the leading company in Sweden interms of MRO work and with the constant development of equipment, more and more risks are discovered that require analysis and minimization. Research has previously been done on how fixture design can be improved ergonomically using virtual environment technology. This is to be able to minimize design and manufacturing costs with regard to the iteration process. This in combination with studies done on musculoskeletal injuries with connections to the MRO industry highlights the need for risk analysis in the MRO industry. The work at STEAS consists of performing risk analysis from a technical, ergonomic and financial perspective on their moving fixtures and chemical processes so that STEAS can then check for possible costs and implement the recommended measures. The analysis methods used are a PHL (Preliminary Hazard List), FMEA (Failure Modes and Effects Analysis) and an ACHB analysis (Accidental Cost &amp; Hypothetical Benefit)

Man-made materials such as fiber-reinforced composites (FRCs) can be tailored for optimum performance in product design applications in terms of strength and weight. The current work is aimed at studying the behaviors of composite laminates based on E-glass CSM (Chopped Strand Mat) or WRM (Woven Roving Mat) plies with a polyester resin for impact protection applications. Detailed mechanical characterization of CSM and WRM laminates till failure is carried out for tensile, compressive and shear loads by varying manufacturing process, number of plies, and laminate thickness. The effect of fiber volume fraction on mechanical properties is shown. The efficacy of CSM and WRM laminates as energy- absorbing countermeasures is studied by performing quasi-static and axial impact tests on cylindrical tubes made of the stated FRCs. In addition to load-displacement and specific energy absorption attributes, failure modes are of interest in such studies. The potential of FRC laminates for protection against projectile impact is investigated by performing low velocity impact perforation tests with a falling tup fitted with an indentor, and medium to high velocity projectile impact tests in a gas gun-based device. The valuable results generated are used for the validation of nonlinear finite element-based CAE (Computer-Aided Engineering) procedures including application of a multi-modal failure criterion for explicit dynamic analysis. The present study not only throws light on complex mechanical behavior of an important class of lightweight materials under static and dynamic loads, but also simulation tools for the design of impact safety countermeasures such as bullet-proof laminates and energy–absorbing components for automotive body structures.