Dynamika a lokalizace antigen prezentujících buněk v MHCII-EGFP knock-in modelech

Pačes, Jan

This doctoral thesis presents an innovative methodological framework for the quantification and phenotyping of immune organs and cell populations in the mouse model. The approach benefits from a transgenic animal model-the MHC II-EGFP knock-in mouse-and combines precise tissue dissection, optimised sample preparation, flow cytometry, stereomicroscopy, three-dimensional imaging using light sheet fluorescence microscopy (LSFM), and data analysis pipelines for quantitative evaluation. Together, these methods form a practical and adaptable toolbox for the histological analysis of immune tissues. The studies included in this thesis demonstrate the application of this toolbox to investigate the distribution, morphology, and interactions of antigen-presenting cell (APC) populations in gastrointestinal and lymphoid tissues. The approach was used to compare different gnotobiotic mouse models, revealing the impact of microbial colonization on immune cell composition and the morphology of the gastrointestinal tract and associated lymphoid tissues. The findings show differences in various quantitative immune parameters among germ-free, oligocolonised, and specific-pathogen-free (SPF) mice. Furthermore, the methods were adapted for the analysis of a primary lymphoid organ-the thymus-highlighting their broader...

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