The lifeact-EGFP mouse is a translationally controlled fluorescent reporter of T cell activation

Jorge Luis Galeano Niño, Szun S. Tay, Jacqueline L.E. Tearle, Jianling Xie, Matt A. Govendir, Daryan Kempe, Jessica Mazalo, Alexander P. Drew, Feyza Colakoglu, Sarah K. Kummerfeld, Christopher G. Proud, Maté Biro

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

It has become increasingly evident that T cell functions are subject to translational control in addition to transcriptional regulation. Here, by using live imaging of CD8+ T cells isolated from the Lifeact-EGFP mouse, we show that T cells exhibit a gain in fluorescence intensity following engagement of cognate tumour target cells. The GFP signal increase is governed by Erk1/2-dependent distal T cell receptor (TCR) signalling and its magnitude correlates with IFN-γ and TNF-α production, which are hallmarks of T cell activation. Enhanced fluorescence was due to increased translation of Lifeact-EGFP protein, without an associated increase in its mRNA. Activation-induced gains in fluorescence were also observed in naive and CD4+ T cells from the Lifeact-EGFP reporter, and were readily detected by both flow cytometry and live cell microscopy. This unique, translationally controlled reporter of effector T cell activation simultaneously enables tracking of cell morphology, F-actin dynamics and activation state in individual migrating T cells. It is a valuable addition to the limited number of reporters of T cell dynamics and activation, and opens the door to studies of translational activity and heterogeneities in functional T cell responses in situ.

Original languageEnglish
Article numberjcs238014
Number of pages15
JournalJournal of Cell Science
Volume133
Issue number5
DOIs
Publication statusPublished - Mar 2020
Externally publishedYes

Keywords

  • Actin
  • Flow cytometry
  • Lifeact
  • Live imaging
  • T cell activation
  • Translation

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