A Dual-Color Bioluminescence Reporter Mouse for Simultaneous in vivo Imaging of T Cell Localization and Function.

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_A6143E0B1670
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A Dual-Color Bioluminescence Reporter Mouse for Simultaneous in vivo Imaging of T Cell Localization and Function.
Journal
Frontiers in immunology
Author(s)
Kleinovink J.W., Mezzanotte L., Zambito G., Fransen M.F., Cruz L.J., Verbeek J.S., Chan A., Ossendorp F., Löwik C.
ISSN
1664-3224 (Electronic)
ISSN-L
1664-3224
Publication state
Published
Issued date
2018
Peer-reviewed
Oui
Volume
9
Pages
3097
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
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 <sup>*</sup> 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.
Keywords
Animals, Benzothiazoles/metabolism, CD4-Positive T-Lymphocytes/metabolism, CD8-Positive T-Lymphocytes/metabolism, Cell Line, Coleoptera/enzymology, Dendritic Cells/immunology, Epitopes, Firefly Luciferin/chemical synthesis, Firefly Luciferin/metabolism, Luciferases, Firefly/metabolism, Luminescent Measurements/methods, Lymph Nodes/cytology, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Mice, Inbred CBA, Mice, Transgenic, NFATC Transcription Factors/metabolism, Ovalbumin/immunology, Spleen/cytology, Vaccination, T cell, activation, bioluminescence, dual-color, imaging, luciferase, mouse, reporter
Pubmed
Web of science
Open Access
Yes
Create date
20/01/2019 15:04
Last modification date
26/10/2019 5:09
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