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Mouse models to deplete or label dendritic cells via genetic manipulation of the Clec9a locus

Van Blijswijk, JM; (2015) Mouse models to deplete or label dendritic cells via genetic manipulation of the Clec9a locus. Doctoral thesis , UCL (University College London). Green open access

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Abstract

Dendritic cells (DCs) play important roles at the interface between innate and adaptive immunity by priming and directing T cell responses. Much of our current knowledge of DC biology has come from mouse models in which DCs can be genetically manipulated, labelled or ablated. Here, novel models are presented using a strategy that targets DC precursors via genetic editing of the Clec9a locus. While validating a novel mouse model to inducibly deplete DCs using diphtheria toxin receptor (DTR) expression driven by Clec9a, it became clear that these Clec9a+/CreROSAiDTR mice suffer from unexpected lymph node (LN) hypocellularity and reduced frequencies of DCs in LNs, even in the absence of diphtheria toxin (DT) injection. This phenotype turned out to be a common feature of other mouse models in which DTR is expressed on DCs (e.g. CD11c-DTR and Langerin-DTR mice) and raises questions about the interpretation of results obtained with such animals. Therefore, in an alternative approach, mice were developed to constitutively lack DCs by expressing the diphtheria toxin alpha (DTA) subunit under control of the Clec9a locus. Unfortunately, these mice still harboured DCs and only showed partial reduction of one DC subset. Finally, seeding of tissues by DC precursors was examined. Clec9a+/CreROSA+/confetti mice were generated in which DC precursors stochastically express one of four fluorescent proteins, which is inherited by its daughter cells. 8- Colour microscopy of tissue sections and histo-cytometry analysis of the images was developed to analyse these mice. This approach will be used to determine how many daughter cells are produced when a single DC precursor seeds the small intestine (clonal burst size), whether these daughters are found among different DC subsets and whether seeding changes during inflammation. In summary, manipulation of the Clec9a locus proves to be an excellent way to study the DC lineage and DC precursor behaviour in the mouse.

Type: Thesis (Doctoral)
Title: Mouse models to deplete or label dendritic cells via genetic manipulation of the Clec9a locus
Event: UCL (University College London)
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Third party copyright material has been removed from ethesis.
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
URI: https://discovery.ucl.ac.uk/id/eprint/1472681
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