The major goal of my study is to understand how the nervous system regulates lung function and disease pathophysiology. Asthma, which is a chronic allergic disease in the lung, has been associated with deregulated airway innervation. The two cell types in the lung that are innervated are airway smooth muscle cells and pulmonary neuroendocrine cells (PNECs). Given that asthma often starts in early childhood, prior research established a neonatal mouse model of allergen exposure to facilitate functional studies of nerves in the development of asthma. Our previous findings showed that allergen exposure to developing, postnatal lungs upregulated levels of neurotrophin 4 (NT4), and caused airway hyperinnervation associated with persistent mucus overproduction. In this work, I describe a novel role of the pulmonary neuroendocrine system in promoting mucus overproduction in early life through deregulated GABAergic signaling. PNECs are the only innervated epithelial cells and express a variety of neuropeptides and bioactive amines. However, how neural innervation affects PNEC secretion and function in disease is not known. Here, I demonstrated that PNECs were the only source of gamma-Aminobutyric acid (GABA) in airways and that GABA hypersecretion from PNECs was required for mucus overproduction following early life allergen exposure. Further, mice lacking NT4 were protected from allergen-induced PNEC hyperinnervation, GABA hypersecretion, and thus mucus overproduction, all which could be rescued with addition of GABA. These findings link PNECs and allergen-induced mucus overproduction through NT4-dependent innervation. Notably, like mice, infant nonhuman primates exhibit PNEC hyperinnervation following early life exposure to ozone and allergens. In addition, I demonstrate that GABA acts in concert with interleukin-13 to induce the proliferation of mucus-producing goblet cells in human airway epithelium cell cultures. Lastly, building upon our previous observations that mast cells contributed to the elevated NT4 levels after allergen exposure, I initiated a research project that investigates the function of a discrete, resident mast cell in: NT4 expression, PNEC innervation, and mucus overproduction. Together, my findings address a novel fundamental role of the neuroendocrine system biology in animal models of asthma. Targeting the nerve–PNEC axis may be a valid treatment strategy for mucus overproduction in asthma. / 2020-06-12T00:00:00Z

<h1>Sammanfattning</h1><h1> </h1><h1>Barn drabbas i allt större utsträckning av allergi i någon form. Det är viktigt att forska kring dess medicinska verkningar på barnen och påverkan av allergin i det dagliga livet. Det forskas mycket idag kring barn och allergi. Läkarna ser en uppgång under senaste decennierna och ingen bättring i sikte. Det kan bero på dagens livsstil och miljötänkande. En större förståelse i dagens samhälle för barnens sjukdom behövs i många sammanhang. Jag använde mig av en kvalitativ intervjumetod för att få reda på barnens kunskaper och tankar kring ämnet. Jag har även gått igenom tidigare forskning kring ämnet. I min litteratur/enkätstudie framkom det att barnen vet mycket om allergi och att de får försaka aktiviteter inom skola/fritid. De barn som hade någon närstående eller kamrat på förskolan med allergi hade goda kunskaper och insikter i hur det var att leva med allergi. Nyckelord: Allergi, Allergen, Immunförsvaret, Pollen och Överkänslighet.</h1><h1> </h1> / Detta är den version som ska gälla, den gamla innehåller fel.

<p>ABSTRACT</p><p>Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.</p>

ABSTRACT Allergy to mustard has been reported for many years, in some cases as severe anaphylactic reactions. Recent studies imply that this allergy is increasing. Three major allergens have been isolated and characterised; Sin a 1 and Sin a 2 in yellow mustard (Sinapis alba), and Bra j 1 in oriental mustard (Brassica juncea). Yellow mustard and black mustard (Brassica nigra) are the most common species in Europe, whereas oriental mustard is more frequent outside Europe. Mustard plants belong to the Brassicaceae/Cruciferae family. Mustard is present as an ingredient in different foods, sauces and spices, often in small amounts. According to the European labelling directives, mustard and products thereof must always be declared. To monitor this regulation, methods need to be developed to detect mustard. Polyclonal antibodies, produced in rabbits, against yellow and black mustard were characterised with immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and immunoblotting. Rocket-immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA) were developed for the detection and quantification of mustard protein. With indirect competitive ELISA a concentration of 156ng mustard protein per ml food extract was detected, which is more than enough to cover the lowest reported reactive doses.

Sammanfattning   Barn drabbas i allt större utsträckning av allergi i någon form. Det är viktigt att forska kring dess medicinska verkningar på barnen och påverkan av allergin i det dagliga livet. Det forskas mycket idag kring barn och allergi. Läkarna ser en uppgång under senaste decennierna och ingen bättring i sikte. Det kan bero på dagens livsstil och miljötänkande. En större förståelse i dagens samhälle för barnens sjukdom behövs i många sammanhang. Jag använde mig av en kvalitativ intervjumetod för att få reda på barnens kunskaper och tankar kring ämnet. Jag har även gått igenom tidigare forskning kring ämnet. I min litteratur/enkätstudie framkom det att barnen vet mycket om allergi och att de får försaka aktiviteter inom skola/fritid. De barn som hade någon närstående eller kamrat på förskolan med allergi hade goda kunskaper och insikter i hur det var att leva med allergi. Nyckelord: Allergi, Allergen, Immunförsvaret, Pollen och Överkänslighet. / Detta är den version som ska gälla, den gamla innehåller fel.

Atopic asthma is characterized by excessive T helper 2 (Th2)-like immunity to allergens in the bronchial mucosa. The Th2-cytokine interleukin (IL)-4 induces IgE production, while the Th2-cytokine IL-5 promotes eosinophilic inflammation in the airways of asthmatics. Most asthmatics are atopic, but a subgroup is non-atopic. We hypothesize that allergen-induced Th2, particularly IL-5, responses can be observed in peripheral blood in both atopic and non-atopic asthmatic children but not in healthy control children. The aim of the present study was to determine IL-4, IL-5, IL-9, IL-10, IL-13 and IFN-γ secretion induced from peripheral blood mononuclear cells (PBMC) by a broad panel of inhalant allergens (timothy, cat, birch, dog and house dust mite) in asthmatic children with and without sensitization. The study included 13 atopic asthmatic, 5 non-atopic asthmatic, and 12 non-atopic non-asthmatic children. PBMC were stimulated with allergens and cytokine production was measured with enzyme-linked immunosorbent assay (ELISA). Higher levels of cat and dog antigen-induced IL-5 release were more commonly observed in both atopic and non-atopic asthmatics than in controls. Children with atopic, but not non-atopic, asthma produced higher levels of allergen-induced IL-4 and IL-9 than controls. Non-atopic asthmatics produced more IL-10 than atopic asthmatics after cat stimulation. High levels of eosinophilia-associated IL-5 responses are induced by cat and dog allergen in both atopic and non-atopic asthmatic children. The Th2 cytokines IL-4 and IL-9 were associated only with atopic asthma, probably due to their IgE-inducing properties.

Proteins are important building blocks of all living organisms. They are composed of a defined sequence of different amino acids, and fold into a specific three-dimensional, ordered structure. The three-dimensional structure largely determines the function of the protein, but protein function always requires motion. Small movements within the protein structure govern the functional properties, and this thesis aims to better understand these discrete protein movements. The motions within the protein structure are governed by thermodynamics, which therefore is useful to predict protein interactions. Nuclear magnetic resonance (NMR) is a powerful tool to study proteins at atomic resolution. Therefore, NMR is the primary method used within this thesis, along with other biophysical techniques such as Fluorescence spectroscopy, Circular Dichroism spectroscopy and in silico modeling. In paper I, NMR in combination with molecular engineering is used to show that the folding of the catalytical subdomains of the enzyme Adenylate kinase does not affect the core of the protein, and thus takes a first step to linking folding, thermodynamic stability and catalysis. In paper II, the structure of the primary allergen from Brazil nut, Ber e 1, is presented along with biophysical measurements that help explain the allergenic potential of the protein. Paper III describes the need for a specific Brazil nut lipid fraction needed to induce an allergenic response. NMR and fluorescence spectroscopy is used to show that there is a direct interaction between Ber e 1 and one or several components in the lipid fraction.

Techn. Universiẗat, Diss., 2005--München.

Identification et caractérisation d'une population d'ILC2 associée à la sévérité de la rhinite allergique et de l'asthmeTrois catégories de cellules lymphoïdes innées (innate lymphoid cells, ILC) ont été décrites récemment sur la base de leurs phénotypes et leurs caractéristiques fonctionnelles : les ILC1, ILC2 et ILC3. Les ILC2 semblent avoir un rôle pro-inflammatoire important dans l’allergie en raison de leur capacité à produire de grandes quantités de cytokines TH2.Pour mieux comprendre le rôle de ces cellules dans l’allergie respiratoire, nous avons comparé les ILC sanguines de patients atteints d’une rhinite allergique associée ou non à un asthme, à celles de sujets non allergiques. Cette étude révèle de multiples différences fonctionnelles entre les ILC circulantes de sujets sains et allergiques. Notamment, la fréquence d’ILC2 exprimant le récepteur aux chimiokines CCR10 est augmentée dans le sang de patients asthmatiques sévères.CCR10 pouvant permettre le recrutement des ILC vers les organes cibles, le rôle des ILC2 CCR10+ dans la physiopathologie de l’asthme a été étudié. Leur présence dans les poumons humains a été observée. Des analyses fonctionnelles et phénotypiques ont révélé que cette sous-population cellulaire était peu activée mais présentait une plasticité leur conférant des caractéristiques proches des ILC1. La déplétion de ces cellules dans un modèle murin d’asthme allergique aggrave l’hyperréactivité bronchique.Les travaux de cette thèse documentent le rôle des ILC dans l’asthme. En particulier, la fréquence sanguine d’ILC2 CCR10+ augmente avec la sévérité de la maladie. Les résultats obtenus dans les modèles animaux suggèrent que ces cellules auraient un rôle bénéfique dans le contrôle de l’asthme. La voie du CCR10 pourrait représenter une nouvelle cible pour le développement de traitements innovants contre l’asthme ou une source prometteuse de biomarqueurs. / Identification and characterization of an ILC2 subset linked to allergic rhinitis and asthma severityInnate lymphoid cells (ILCs) have been classified into ILC1, ILC2 and ILC3 subsets based on their respective phenotypes and functions. Considering the strong ability of ILC2s to produce TH2 cytokines, these cells likely play a significant role in allergic diseases.To better understand the role of these cells in respiratory allergies, we compared blood ILCs from allergic patients with or without asthma to non-allergic individuals. Together our results show multiple functional differences between ILC from allergic and healthy subjects. In particular, ILC2s expressing the chemokine receptor CCR10 are specifically enriched in the blood of patients with severe allergic asthma.Considering that CCR10 could allow the recruitment of ILCs to target organs, the role of CCR10+ ILC2s in asthma physiopathology has been studied. This ILC2 subtype is present in human lungs. Functional and phenotypic analyses revealed that these cells are less activated than other ILC2s and show ILC1-like properties. CCR10+ ILC2s depletion in a mouse model of allergic asthma exacerbate airway hyperreactivity.Together, this work documents the role of ILCs in asthma. Specifically, circulating CCR10+ ILC2 frequency increases with asthma severity. The results obtained in mouse models suggest that these cells could have a beneficial role in asthma control. CCR10 pathway could represent a new target to elaborate breakthrough treatments against asthma or a source of promising biomarkers.

Ph.D. THESIS TITLE Nutritional Biology of Synanthropic Mites (Acari: Acaridida) ABSTRACT Several attempts to describe the nutritional biology of acaridid mites were undertaken, however full understanding of these processes remains incomplete. The objective of this Ph.D. thesis was to expand our knowledge concerning digestive physiology of stored product and house dust mites and to apply this knowledge to their nutritional biology. The research approach adopted in this Ph.D. thesis includes in vitro characterization of enzymatic activity in whole mite extracts (WME) and spent growth medium extracts (SGME), evaluation of the enzyme activities with respect to the gut physiological pH, enzyme inhibition experiments, in vivo localization of enzyme activities in the mite gut, determination of effects of nutrient or antifeedant additives in experimental diets on mite population growth and determination of the feeding preferences of synanthropic mites as assessed by in vitro and in vivo analyses. The gut contents of twelve species of synanthropic acaridid mites were determined to be within a pH range of 4 to 7 and showed a pH gradient from the anterior to the posterior midgut. The pH in digestive tract of synanthropic acaridid mites corresponds to the activity of proteases, α-glucosidases, α-amylases and...