@phdthesis{Mayer2006,
  author    = {Mayer, Katrin Doris},
  title     = {Visualization of type I immunity using bicistronic IFN-gamma reporter mice in vitro and in vivo},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-19415},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2006},
  abstract  = {Typ I Immunantworten, wie z.B. gegen Influenza Virus, Sendai Virus aber auch gegen intrazellul{\"a}re Erreger wie Toxoplasma gondii sind klassischerweise durch robuste IFN-\&\#947; Expression gekennzeichnet. Th1 und CD8+ Effektor T Zellen z{\"a}hlen zu den Hauptproduzenten von IFN-\&\#947;. Im Zusammenhang mit Autoimmunerkrankungen, Immunpathologie aber auch Impfstoffentwicklung, ist es {\"u}beraus wichtig die Regulierung von IFN-\&\#947; zu verstehen. In der vorliegenden Arbeit wurde die IFN-\&\#947; Expression von CD4+ und CD8+ T Zellen detailliert charakterisiert. Des Weiteren wurde die Rolle des IFN-\&\#947; Rezeptors f{\"u}r die IFN-\&\#947; Expression von T Zellen untersucht. Unter Zuhilfenahme von bicistronischen IFN-\&\#947;-eYFP Reporter M{\"a}usen, welche die direkte Identifizierung und Isolierung von vitalen IFN-\&\#947; exprimierenden Zellen erm{\"o}glichen, wurde die Expression von IFN-\&\#947; in vitro und in vivo, nach Infektion mit den bereits erw{\"a}hnten Erregern,visualisiert. Die Expression des IFN-\&\#947;-eYFP Reporters zeichnete sich, sowohl in vitro als auch in vivo nach Infektion, durch ein {\"a}ußerst heterogenes Fluoreszenzspektrum aus. Die Helligkeit der Reporter Fluoreszenz korrelierte positiv mit der Menge an IFN-\&\#947; Transkripten und mit der Menge des sekretierten IFN-\&\#947; Proteins nach Stimulierung. Die Helligkeit des Reporters reflektierte das Potenzial zur IFN-\&\#947; Produktion, die eigentliche Sekretion war jedoch weitgehend abh{\"a}ngig von zus{\"a}tzlicher Stimulierung durch Antigen. Des Weiteren korrelierte die Helligkeit des Reporters mit der zunehmenden Produktion von weiteren proinflammatorischen Zytokinen und Chemokinen. Hoch fluoreszente Zellen exprimierten zudem vermehrt Marker auf ihrer Oberflache, die auf akute Aktivierung hinweisen. Die am hellsten eYFP fluoreszierenden Zellen waren im Allgemeinen weiter ausdifferenziert und ihre Pr{\"a}senz war auf bestimmte Organe beschr{\"a}nkt. Die anatomische Begrenzung wurde durch den Erreger bestimmt. IFN-\&\#947; exprimierende Zellen wurden nach Infektion mit Sendai Virus oder Toxoplasma gondii in IFN-\&\#947; Rezeptor defizienten Reporter M{\"a}usen generiert. Die Frequenz und die Helligkeit der eYFP Reporter Expression waren jedoch ver{\"a}ndert. Experimente mit dualen Knochenmarks-Chim{\"a}ren M{\"a}usen, welche mit Wild-Typ und IFN-\&\#947; Rezeptor defizientem Knochenmark rekonstituiert wurden, ergaben eine T Zell-intrinsische Abh{\"a}ngigkeit von IFN-\&\#947; Rezeptor vermittelten Signalen f{\"u}r die Expression von IFN-\&\#947;. Die Helligkeit des Reporters dagegen wurde unabh{\"a}ngig von dem IFN-\&\#947; Rezeptor reguliert. Abschließend wurde ein Modell f{\"u}r die Expression von IFN-\&\#947; in CD4+ und CD8+ T Zellen entwickelt. Zusammenfassend f{\"u}hren diese Ergebnisse zu dem Schluss, dass die Expression von IFN-\&\#947; in CD4+ und CD8+ T Zellen und nach viraler oder parasit{\"a}rer Infektion unterschiedlich reguliert wird. Zus{\"a}tzlich wurde gezeigt, dass der IFN-\&\#947; Rezeptor an der \&\#65279;Modulation der IFN-\&\#947; Expression beteiligt ist.},
  subject      = {Interferon <gamma->},
  language  = {en}
}
@phdthesis{Bucher2018,
  author    = {Bucher, Hannes},
  title     = {Pre-clinical modeling of viral- and bacterial-induced exacerbations of chronic obstructive pulmonary disease},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-144368},
  school      = {Universit{\"a}t W{\"u}rzburg},
  pages     = {XIII, 105},
  year      = {2018},
  abstract  = {Chronic Obstructive Pulmonary Disease (COPD) exacerbations are a considerable reason for increased morbidity and mortality in patients. Infections with influenza virus (H1N1), respiratory syncytial virus (RSV) or nontypeable Haemophilus influenzae (NTHi) are important triggers of exacerbations. To date, no treatments are available which can stop the progression of COPD. Novel approaches are urgently needed. Pre-clinical models of the disease are crucial for the development of novel therapeutic options. In order to establish pre-clinical models which mimic aspects of human COPD exacerbations, mice were exposed to cigarette smoke (CS) and additionally infected with H1N1, RSV and/or NTHi. Clinically relevant treatments such as the corticosteroids Fluticasone propionate and Dexamethasone, the phosphodiesterase-4 (PDE-4) inhibitor Roflumilast and the long-acting muscarinic receptor antagonist Tiotropium were tested in the established models. Furthermore, a novel treatment approach using antibodies (Abs) directed against IL-1α, IL-1β or IL-1R1 was examined in the established CS/H1N1 model. Levels of IFN-γ, IL-1β, IL-2, IL-6, KC, TNF-α, RANTES, IL-17, MCP-1, MIP 1α and MIP-1β were measured in lung homogenate. Numbers of total cells, neutrophils and macrophages were assessed in bronchoalveolar lavage (BAL) fluid. Hematoxylin- and eosin- (H\&E-) stained lung slices were analyzed to detect pathological changes. Quantitative polymerase-chain-reaction (qPCR) was used to investigate gene expression of ICAM-1 and MUC5 A/C. The viral/bacterial load was investigated in lung homogenate or BAL fluid. In addition to the in vivo studies, the effects of the above mentioned treatments were investigated in vitro in H1N1, RSV or NTHi-infected (primary) human bronchial epithelial cells using submerged or air-liquid-interface (ALI) cell culture systems. Four pre-clinical models (CS/H1N1, CS/RSV, CS/NTHi, CS/H1N1/NTHi) were established depicting clinically relevant aspects of COPD exacerbations such as increased inflammatory cells and cytokines in the airways and impaired lung function. In the CS/H1N1 model, Tiotropium improved lung function and was superior in reducing inflammation in comparison to Fluticasone or Roflumilast. Moreover, Fluticasone increased the loss of body-weight, levels of IL-6, KC and TNF-α and worsened lung function. In CS/RSV-exposed mice Tiotropium but not Fluticasone or Roflumilast treatment reduced neutrophil numbers and IL-6 and TNF α levels in the lung. The viral load of H1N1 and RSV was significantly elevated in CS/virus-exposed mice and NCI-H292 cells after Fluticasone and Dexamethasone treatment. The results from these studies demonstrate that Tiotropium has anti-inflammatory effects on CS/virus-induced inflammation and might help to explain the observed reduction of exacerbation rates in Tiotropium-treated COPD patients. Furthermore, the findings from this work indicate that treatment with Fluticasone or Dexamethasone might not be beneficial to reduce inflammation in the airways of COPD patients and supports clinical studies that link treatment with corticosteroids to an increased risk for pneumonia. Testing of anti-IL-1α, anti-IL-1β or anti-IL-1R1 Abs in the CS/H1N1 model suggests that, in line with clinical data, antagonization of IL-1β is not sufficient to reduce pulmonary inflammation and indicates a predominant role of IL-1α in CS/virus-induced airway inflammation. In line with the in vivo findings, anti-IL-1α but not anti-IL-1β Abs reduced levels of TNF-α and IL-6 in H1N1-infected primary human bronchial epithelial ALI cell culture. Blocking the IL-1R1 provided significant inhibitory effects on inflammatory cells in vivo but was inferior compared to inhibiting both its soluble ligands IL-1α and IL-1β. Concomitant usage of Abs against IL-1α/IL-1β revealed strong effects and reduced total cells, neutrophils and macrophages. Additionally, levels of KC, IL-6, TNF-α, MCP-1, MIP-1α and MIP-1β were significantly reduced and ICAM-1 mRNA expression was attenuated. These results suggest that combined inhibition of IL-1α/IL-1β might be beneficial to reduce inflammation and exacerbations in COPD patients. Moreover, combined targeting of both IL-1α/IL-1β might be more efficient compared to inhibition of the IL-1R1. As in the CS/virus models, corticosteroid treatment failed to reduce inflammatory cells in the CS/NTHi and CS/H1N1/NTHi models, increased the loss of body-weight and the bacterial load. Furthermore, Roflumilast administration had no significant effects on cell counts or cytokines. However, it improved compliance in the CS/NTHi model. Treatment with Azithromycin reduced the bacterial load in the CS/NTHi model and reduced numbers of total cells, neutrophils, macrophages and levels of KC and TNF-α in the CS/H1N1/NTHi model. In conclusion, the established CS/H1N1, CS/RSV, CS/NTHi, CS/H1N1/NTHi models depict clinically relevant aspects of human COPD exacerbations in mice and provide the opportunity to investigate underlying disease mechanisms and to test novel therapies.},
  subject      = {Obstruktive Ventilationsst{\"o}rung},
  language  = {en}
}