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Characterization of ex vivo immunostimulated dendritic cells in murine lung tissue

 
: Brandstetter, Ronja Lorena
: Braun, Armin; Schwinzer, Reinhard

Hannover, 2014, V, 49 pp.
Hannover, Medizinische Hochschule, Master Thesis, 2014
English
Master Thesis
Fraunhofer ITEM ()

Abstract
Although there has been excessive research in the field of inflammatory airway diseases throughout the last decades, there are no approaches to inhibit the causes of asthma. Therapy of asthma remains symptomatic and there is no solution in sight to prevent onset of disease. On the one hand, the field is hampered by inconclusive in vitro experiments, omitting the complex environment provided by the lung tissue. On the other hand, the majority of current studies are based on animal testing, which is one of the most contentious issues in science.
We applied the ex vivo method of precision-cut lung slices (PCLS) to circumvent lack of biological coherence, but at the same time maintaining the ability to assess different cell types by flow cytometry (FC). We focused on dendritic cells (DCs) and their migration in response to exogenous stimuli.
The key players of the innate immune system during the induction phase of atopic allergic diseases are DCs. In order to study DCs in lung context dependent manner, they have to be embedded in a structural network of a variety of haematopoietic and mesenchymal cell types contributing to their function. In this work we confirm that the composition of murine PCLS provide a cellular context in which DCs are embedded. We characterized CD11c+ cells from LPS-stimulated PCLS regarding their expression of the co-stimulatory molecule CD86 and the antigen-presenting molecule MHCII. Additionally to the determined functional competence of immune-stimulated DCs within PCLS, we investigated their migration potential along a CCL19 gradient into an extracellularmatrix. We demonstrated that LPS does not significantly modulate expression of CD86, MHCII or CCR7 in CD11c+, in the presence of DQTM-OVA, but it appeared to have an influence on migration of DCs.
We thus provided a method to characterise and assess the phenotype of PCLS derived cells by FC. Therefore tissue embedded cells can be subjected to a treatment of choice and subsequently analyzed for their phenotype, functional properties and activation status.

: http://publica.fraunhofer.de/documents/N-317983.html