{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T04:47:41Z","timestamp":1791002861591,"version":"4.1.0"},"reference-count":29,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2022,11,21]],"date-time":"2022-11-21T00:00:00Z","timestamp":1668988800000},"content-version":"vor","delay-in-days":324,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Career Development","award":["MR\/V009052\/1"],"award-info":[{"award-number":["MR\/V009052\/1"]}]},{"name":"Career Development","award":["C61638\/A27112"],"award-info":[{"award-number":["C61638\/A27112"]}]},{"DOI":"10.13039\/501100000268","name":"Biotechnology and Biological Sciences Research Council","doi-asserted-by":"publisher","award":["BB\/J013951\/2"],"award-info":[{"award-number":["BB\/J013951\/2"]}],"id":[{"id":"10.13039\/501100000268","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,1,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>In lymphocytes, Nr4a gene expression is specifically regulated by antigen receptor signalling, making them ideal targets for use as distal T cell receptor (TCR) reporters. Nr4a3-Timer of cell kinetics and activity (Tocky) mice are a ground-breaking tool to report TCR-driven Nr4a3 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 Nr4a1\/Nur77 may be a more sensitive gene to distal TCR signals compared to Nr4a3, 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 Nr4a1 to lower TCR signal strengths.<\/jats:p>","DOI":"10.1093\/discim\/kyac009","type":"journal-article","created":{"date-parts":[[2022,11,21]],"date-time":"2022-11-21T10:09:49Z","timestamp":1669025389000},"source":"Crossref","is-referenced-by-count":29,"title":["Nur77-Tempo mice reveal T cell steady state antigen recognition"],"prefix":"10.1093","volume":"1","author":[{"given":"Thomas A E","family":"Elliot","sequence":"first","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma K","family":"Jennings","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A J","family":"Lecky","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Rouvray","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gillian M","family":"Mackie","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa","family":"Scarfe","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lozan","family":"Sheriff","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masahiro","family":"Ono","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, Imperial College London , London, SW7 2AZ , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kendle M","family":"Maslowski","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0071-1163","authenticated-orcid":false,"given":"David","family":"Bending","sequence":"additional","affiliation":[{"name":"Institute of Immunology and Immunotherapy, College of Medical and Dental Sciences, University of Birmingham , Birmingham B15 2TT , UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,11,21]]},"reference":[{"key":"2022122213273760200_CIT0001","doi-asserted-by":"crossref","first-page":"208","DOI":"10.4049\/immunohorizons.1800032","article-title":"Peptide antigen concentration modulates digital NFAT1 activation in primary mouse naive CD8+ T cells as measured by flow cytometry of isolated cell nuclei","volume":"2","author":"Gallagher","year":"2018","journal-title":"ImmunoHorizons"},{"key":"2022122213273760200_CIT0002","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.cell.2008.11.051","article-title":"Digital Signaling and hysteresis characterize Ras activation in lymphoid cells","volume":"136","author":"Das","year":"2009","journal-title":"Cell"},{"key":"2022122213273760200_CIT0003","doi-asserted-by":"crossref","first-page":"e2025825118","DOI":"10.1073\/pnas.2025825118","article-title":"Hierarchy of signaling thresholds downstream of the T cell receptor and the Tec kinase ITK","volume":"118","author":"Gallagher","year":"2021","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022122213273760200_CIT0004","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1111\/j.1365-2567.2011.03547.x","article-title":"T-cell receptor affinity in thymic development","volume":"135","author":"Moran","year":"2012","journal-title":"Immunology"},{"key":"2022122213273760200_CIT0005","doi-asserted-by":"crossref","first-page":"eaas9103","DOI":"10.1126\/sciimmunol.aas9103","article-title":"TCR signal strength controls the differentiation of CD4+ effector and memory T cells","volume":"3","author":"Snook","year":"2018","journal-title":"Sci Immunol"},{"key":"2022122213273760200_CIT0006","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.4049\/jimmunol.1901167","article-title":"TCR signal strength and antigen affinity regulate CD8+ memory T cells","volume":"205","author":"Solouki","year":"2020","journal-title":"J Immunol"},{"key":"2022122213273760200_CIT0007","doi-asserted-by":"crossref","first-page":"2481","DOI":"10.1016\/j.immuni.2021.08.020","article-title":"Antigen and checkpoint receptor engagement recalibrates T cell receptor signal strength","volume":"54","author":"Elliot","year":"2021","journal-title":"Immunity"},{"key":"2022122213273760200_CIT0008","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1038\/nature02284","article-title":"Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1","volume":"427","author":"Matloubian","year":"2004","journal-title":"Nature"},{"key":"2022122213273760200_CIT0009","doi-asserted-by":"crossref","first-page":"2412","DOI":"10.1084\/jem.20190414","article-title":"Resident memory CD8 T cells persist for years in human small intestine","volume":"216","author":"Bartolom\u00e9-Casado","year":"2019","journal-title":"J Exp Med"},{"key":"2022122213273760200_CIT0010","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1084\/jem.20181365","article-title":"CD4+ resident memory T cells dominate immunosurveillance and orchestrate local recall responses","volume":"216","author":"Beura","year":"2019","journal-title":"J Exp Med"},{"key":"2022122213273760200_CIT0011","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.imlet.2022.02.009","article-title":"Heterogeneity of tissue resident memory T cells","volume":"245","author":"Konjar","year":"2022","journal-title":"Immunol Lett"},{"key":"2022122213273760200_CIT0012","doi-asserted-by":"crossref","first-page":"4866","DOI":"10.4049\/jimmunol.1200402","article-title":"Antigen-independent differentiation and maintenance of effector-like resident memory T cells in tissues","volume":"188","author":"Casey","year":"2012","journal-title":"J Immunol"},{"key":"2022122213273760200_CIT0013","doi-asserted-by":"crossref","first-page":"1140","DOI":"10.1038\/s41590-021-01004-1","article-title":"Discrete tissue microenvironments instruct diversity in resident memory T cell function and plasticity","volume":"22","author":"Christo","year":"2021","journal-title":"Nat Immunol"},{"key":"2022122213273760200_CIT0014","doi-asserted-by":"crossref","first-page":"7037","DOI":"10.1073\/pnas.1202288109","article-title":"Long-lived epithelial immunity by tissue-resident memory T (T RM) cells in the absence of persisting local antigen presentation","volume":"109","author":"Mackay","year":"2012","journal-title":"Proc Natl Acad Sci USA"},{"key":"2022122213273760200_CIT0015","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1038\/nature04606","article-title":"CD69 acts downstream of interferon-\u03b1\/\u03b2 to inhibit S1P1 and lymphocyte egress from lymphoid organs","volume":"440","author":"Shiow","year":"2006","journal-title":"Nature"},{"key":"2022122213273760200_CIT0016","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1111\/j.1600-065X.1994.tb00894.x","article-title":"Identification of genes induced during apoptosis in T lymphocytes","volume":"142","author":"Osborne","year":"1994","journal-title":"Immunol Rev"},{"key":"2022122213273760200_CIT0017","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1084\/jem.20110308","article-title":"T cell receptor signal strength in Treg and iNKT cell development demonstrated by a novel fluorescent reporter mouse","volume":"208","author":"Moran","year":"2011","journal-title":"J Exp Med"},{"key":"2022122213273760200_CIT0018","doi-asserted-by":"crossref","first-page":"657","DOI":"10.4049\/jimmunol.1601301","article-title":"Endogenous Nur77 is a specific indicator of antigen receptor signaling in human T and B cells","volume":"198","author":"Ashouri","year":"2017","journal-title":"J Immunol"},{"key":"2022122213273760200_CIT0019","doi-asserted-by":"crossref","first-page":"108328","DOI":"10.1016\/j.celrep.2020.108328","article-title":"Nr4a1 and Nr4a3 reporter mice are differentially sensitive to T cell receptor signal strength and duration","volume":"33","author":"Jennings","year":"2020","journal-title":"Cell Rep"},{"key":"2022122213273760200_CIT0020","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1038\/nature11311","article-title":"Endogenous antigen tunes the responsiveness of naive B cells but not T cells","volume":"489","author":"Zikherman","year":"2012","journal-title":"Nature"},{"key":"2022122213273760200_CIT0021","doi-asserted-by":"crossref","first-page":"2931","DOI":"10.1083\/jcb.201711048","article-title":"A timer for analyzing temporally dynamic changes in transcription during differentiation in vivo","volume":"217","author":"Bending","year":"2018","journal-title":"J Cell Biol"},{"key":"2022122213273760200_CIT0022","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1038\/nchembio.138","article-title":"Monomeric fluorescent timers that change color from blue to red report on cellular trafficking","volume":"5","author":"Subach","year":"2009","journal-title":"Nat Chem Biol"},{"key":"2022122213273760200_CIT0023","doi-asserted-by":"crossref","first-page":"100284","DOI":"10.1016\/j.xpro.2020.100284","article-title":"Application of dual Nr4a1-GFP Nr4a3-Tocky reporter mice to study T cell receptor signaling by flow cytometry","volume":"2","author":"Jennings","year":"2021","journal-title":"STAR Protocols"},{"key":"2022122213273760200_CIT0024","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1007\/978-1-0716-0338-3_13","article-title":"Isolation, characterization, and culture of intestinal intraepithelial lymphocytes.","volume":"2121","author":"James","year":"2020","journal-title":"Methods Mol Biol"},{"key":"2022122213273760200_CIT0025","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1038\/nri3155","article-title":"Selection of regulatory T cells in the thymus","volume":"12","author":"Hsieh","year":"2012","journal-title":"Nat Rev Immunol"},{"key":"2022122213273760200_CIT0026","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1016\/j.it.2017.06.010","article-title":"Tonic signals: why do lymphocytes bother?","volume":"38","author":"Myers","year":"2017","journal-title":"Trends Immunol"},{"key":"2022122213273760200_CIT0027","doi-asserted-by":"crossref","first-page":"e70055","DOI":"10.7554\/eLife.70055","article-title":"Tissue environment, not ontogeny, defines murine intestinal intraepithelial T lymphocytes.","volume":"10","author":"Brenes","year":"2021","journal-title":"Elife"},{"key":"2022122213273760200_CIT0028","first-page":"7","article-title":"Tissue-resident memory T cell reactivation by diverse antigen-presenting cells imparts distinct functional responses","volume":"21","author":"Low","year":"2020","journal-title":"J Exp Med"},{"key":"2022122213273760200_CIT0029","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1016\/j.immuni.2019.09.006","article-title":"Butyrophilin-like 3 directly binds a human V\u03b34+ T cell receptor using a modality distinct from clonally-restricted antigen","volume":"51","author":"Willcox","year":"2019","journal-title":"Immunity"}],"container-title":["Discovery Immunology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/discovimmunology\/advance-article-pdf\/doi\/10.1093\/discim\/kyac009\/47159753\/kyac009.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/discovimmunology\/article-pdf\/1\/1\/kyac009\/48349365\/kyac009.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/discovimmunology\/article-pdf\/1\/1\/kyac009\/48349365\/kyac009.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,22]],"date-time":"2022-12-22T13:28:11Z","timestamp":1671715691000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/discovimmunology\/article\/doi\/10.1093\/discim\/kyac009\/6835142"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,1]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2022,1,1]]}},"URL":"https:\/\/doi.org\/10.1093\/discim\/kyac009","relation":{},"ISSN":["2754-2483"],"issn-type":[{"value":"2754-2483","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2022,1,1]]},"published":{"date-parts":[[2022,1,1]]},"article-number":"kyac009"}}