Elliot Thomas A E, Jennings Emma K, Lecky David A J, Rouvray Sophie, Mackie Gillian M, Scarfe Lisa, Sheriff Lozan, Ono Masahiro, Maslowski Kendle M, Bending David
Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham, Birmingham B15 2TT, UK.
Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
Discov Immunol. 2022 Dec 22;1(1):kyac009. doi: 10.1093/discim/kyac009.
In lymphocytes, gene expression is specifically regulated by antigen receptor signalling, making them ideal targets for use as distal T cell receptor (TCR) reporters. -Timer of cell kinetics and activity (Tocky) mice are a ground-breaking tool to report TCR-driven expression using Fluorescent Timer protein (FT). FT undergoes a time-dependent shift in its emission spectrum following translation, allowing for the temporal reporting of transcriptional events. Our recent work suggested that /Nur77 may be a more sensitive gene to distal TCR signals compared to , so we, therefore, generated Nur77-Timer-rapidly-expressed-in-lymphocytes (Tempo) mice that express FT under the regulation of Nur77. We validated the ability of Nur77-Tempo mice to report TCR and B cell receptor signals and investigated the signals regulating Nur77-FT expression. We found that Nur77-FT was sensitive to low-strength TCR signals, and its brightness was graded in response to TCR signal strength. Nur77-FT detected positive selection signals in the thymus, and analysis of FT expression revealed that positive selection signals are often persistent in nature, with most thymic Treg expressing FT Blue. We found that active TCR signals in the spleen are low frequency, but CD69 lymphoid T cells are enriched for FT Blue Red T cells, suggesting frequent TCR signalling. In non-lymphoid tissue, we saw a dissociation of FT protein from CD69 expression, indicating that tissue residency is not associated with tonic TCR signals. Nur77-Tempo mice, therefore, combine the temporal dynamics from the Tocky innovation with increased sensitivity of to lower TCR signal strengths.
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