A platform for research: civil engineering, architecture and urbanism
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Characterization of the murine pulmonary phagocytic network during Aspergillus fumigatus infection ; Charakterisierung des murinen pulmonalen phagoyztischen Netzwerks während einer Aspergillus fumigatus Infektion
Aspergillus fumigatus is a ubiquitous, airborne mold and a common threat for immunosuppressed patients as it can induce invasive aspergillosis, characterized by a tremendously high mortality rate. Immunocompetent individuals on the other hand, encounter this infection very efficiently due to the phagocytic action of different immune cell types such as alveolar macrophages, neutrophil granulocytes and dendritic cells we believe, that beside these professional phagocytes, “unprofessional” phagocytic cells, such as alveolar epithelial type I and II cells (AEC) also constitute a central element for encountering fungal invasion. Although many studies have deeply analyzed the interaction between professional phagocytes and A. fumigatus, just few works have concentrated on the fungal interaction with alveolar epithelial cells. With this ground, we started with analyzing the proteome regulation of AECII after pulmonary A. fumigatus infection. We started with the successful establishment of a negative, magnetic isolation protocol to isolate primary murine AECII with purity greater than 90 %. Using this method, we isolated AECII from infected murine lungs and characterized their proteomic constitution by liquid chromatography mass spectrometry (LCMS) in comparison to AECII from uninfected animals. From the resulting list of significantly regulated proteins the protein with the highest up-regulation was chosen for further investigation in the context of an A. fumigatus infection. By additional experimental approaches, such as qRT-PCR, Western Blot and Immunohistology we could first of all confirm the initially detected high level of up-regulation. Subsequently, we employed a knock-out mouse model for the protein to investigate its significance under infection conditions. Addressing different parameters, such as survival, colony forming units (CFU), Neutrophil recruitment to the lungs of the infected animals and TNF- α secretion level in bronchoalveolar lavage fluid, we unfortunately could not find a different behavior of ...
Characterization of the murine pulmonary phagocytic network during Aspergillus fumigatus infection ; Charakterisierung des murinen pulmonalen phagoyztischen Netzwerks während einer Aspergillus fumigatus Infektion
Aspergillus fumigatus is a ubiquitous, airborne mold and a common threat for immunosuppressed patients as it can induce invasive aspergillosis, characterized by a tremendously high mortality rate. Immunocompetent individuals on the other hand, encounter this infection very efficiently due to the phagocytic action of different immune cell types such as alveolar macrophages, neutrophil granulocytes and dendritic cells we believe, that beside these professional phagocytes, “unprofessional” phagocytic cells, such as alveolar epithelial type I and II cells (AEC) also constitute a central element for encountering fungal invasion. Although many studies have deeply analyzed the interaction between professional phagocytes and A. fumigatus, just few works have concentrated on the fungal interaction with alveolar epithelial cells. With this ground, we started with analyzing the proteome regulation of AECII after pulmonary A. fumigatus infection. We started with the successful establishment of a negative, magnetic isolation protocol to isolate primary murine AECII with purity greater than 90 %. Using this method, we isolated AECII from infected murine lungs and characterized their proteomic constitution by liquid chromatography mass spectrometry (LCMS) in comparison to AECII from uninfected animals. From the resulting list of significantly regulated proteins the protein with the highest up-regulation was chosen for further investigation in the context of an A. fumigatus infection. By additional experimental approaches, such as qRT-PCR, Western Blot and Immunohistology we could first of all confirm the initially detected high level of up-regulation. Subsequently, we employed a knock-out mouse model for the protein to investigate its significance under infection conditions. Addressing different parameters, such as survival, colony forming units (CFU), Neutrophil recruitment to the lungs of the infected animals and TNF- α secretion level in bronchoalveolar lavage fluid, we unfortunately could not find a different behavior of ...
Characterization of the murine pulmonary phagocytic network during Aspergillus fumigatus infection ; Charakterisierung des murinen pulmonalen phagoyztischen Netzwerks während einer Aspergillus fumigatus Infektion
Seddigh, Pegah (author) / Gunzer, Matthias
2017-10-06
Theses
Electronic Resource
English
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