MHC Multimer-Binding CD8 T Cells: Optimized Detection, Automated Analysis and Identification of Antigen Specific CD8 T Cells in Narcolepsy Type 1
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MHC multimer-binding CD8 T cells: Optimized detection, automated analysis and identification of antigen specific CD8 T cells in Narcolepsy type 1 PhD Thesis Natasja Wulff Pedersen October 2018 DTU Nanotech Department of Micro- and Nanotechnology 1015 106 105 107 1015 β α ββ β α α monomers to a streptavidin -conjugated fluorochrome . Using the fluorescent label, CD8 T cells that had recognized the pMHC complex and bound the tetramer could be identified by flow cytometry. Figure 2. Single interactions between TCR and pMHC are unstable and not sufficiently strong to allow fluorescence based detection, whereas the simultaneous binding between several molecules increases the avidity of the interaction enough for detection to be possible. Modified from \citep{Davis2011}. This strategy offered an attractive alternative to conventional methods of detecting antigen specific T cells, as for instance the elispot assay, limiting dilution assay and intracellular cytokine staining, that all rely on the functionality of antigen specific T cells. Though these methods provide an important .