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Abstract
Non-invasive imaging technologies to visualize the location and functionality of T cells are of great value in immunology. Here, we describe the design and generation of a transgenic mouse in which all T cells constitutively express green-emitting click-beetle luciferase (CBG99) while expression of the red-emitting firefly luciferase (PpyRE9) is induced by Nuclear Factor of Activated T cells (NFAT) such as during T cell activation, which allows multicolor bioluminescence imaging of T cell location and function. This dual-luciferase mouse, which we named TbiLuc, showed high constitutive luciferase expression in lymphoid organs such as lymph nodes and the spleen. Ex vivo purified CD8+ and CD4+ T cells both constitutively expressed luciferase, whereas B cells showed no detectable signal. We cross-bred TbiLuc mice to T cell receptor-transgenic OT-I mice to obtain luciferase-expressing naïve CD8+ T cells with defined antigen-specificity. TbiLuc*OT-I T cells showed a fully antigen-specific induction of the T cell activation-dependent luciferase. In vaccinated mice, we visualized T cell localization and activation in vaccine-draining lymph nodes with high sensitivity using two distinct luciferase substrates, D-luciferin and CycLuc1, of which the latter specifically reacts with the PpyRE9 enzyme. This dual-luciferase T cell reporter mouse can be applied in many experimental models studying the location and functional state of T cells.
Highlights
Bioluminescence imaging (BLI) is an optical molecular imaging technique based on the emission of light produced by luciferase enzymes expressed in cells or animals
We report on the design and development of a dualluciferase T cell transgenic mouse, called TbiLuc, as a novel tool for non-invasive imaging of T cells
The dual-luciferase construct allows live visualization of both the location and the activation status of T cells, which we established in antigen-specific T cell activation studies both in vitro and in vivo
Summary
Bioluminescence imaging (BLI) is an optical molecular imaging technique based on the emission of light produced by luciferase enzymes expressed in cells or animals. Besides functioning as a quantitative measure for cell number, BLI provides information on cell viability as the light-producing. Luciferase-transgenic reporter mouse strains can be developed using similar constructs, requiring more time and effort but becoming a source of luciferase-expressing cells without the need for further modification. Such transgenic models are especially valuable if the cells of interest are to be studied in their natural, unmodified state
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