Fluorescent tagged NK cell receptors expressed in vivo using retrogenic mice to study the delivery of signals in NK cells

Abstract

Event Abstract Back to Event Fluorescent tagged NK cell receptors expressed in vivo using retrogenic mice to study the delivery of signals in NK cells David Tomaz1, Nadia Guerra2, Julian Dyson3 and Keith Gould1* 1 Imperial College London, Department of Immunology, United Kingdom 2 Imperial College London, Department of Life Sciences, United Kingdom 3 Imperial College London, Division of Immunology and Inflammation, United Kingdom NK cells display both activating and inhibitory receptors which recognize different ligands, and it is the integration of signals from these two classes of receptors that ultimately determines NK cell effector activity. Nevertheless, how exactly the signal integration occurs is still not well understood. Our approach is to employ retrogenic mice to generate authentic primary NK cells expressing two fluorescent tagged receptors, NKG2D (short or long isoform) and Ly49A, prototypical activating and inhibitory NK cell receptors, respectively. The aim is to use NKG2D-GFP/Ly49A-RFP Forster resonance energy transfer (FRET) to study the co-localisation of these receptors in synapses at NK-target cell-cell contact interfaces. Also, we will test the consequences of altering ligand dimensions in the synapse formation and NK cell activation. Our results show that GFP-NKG2D (short and long isoform) interacts with both adaptor molecules DAP10 and DAP12 in vitro, allowing efficient cell surface expression of fluorescent NKG2D. Also, we reconstituted irradiated C57BL/6 recipients with GFP-NKG2D transduced bone marrow cells from NKG2D-deficient C57BL/6 donor mice. Two weeks post reconstitution, we detected cell surface expression of GFP-NKG2D predominantly in NK cells. The functionality of the fluorescent receptors and their interaction with the adaptor proteins DAP10 and DAP12 are now being investigated in NK cells generated in vivo. Data on the functionality of the fluorescent receptors and the generation of NK cells in retrogenic mice will be presented. Keywords: NK cells, innate immunity, NKG2D, ly49a, retrogenic mice Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Immune receptors and signaling Citation: Tomaz D, Guerra N, Dyson J and Gould K (2013). Fluorescent tagged NK cell receptors expressed in vivo using retrogenic mice to study the delivery of signals in NK cells. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01194 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 22 Jul 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Keith Gould, Imperial College London, Department of Immunology, London, United Kingdom, k.gould@ic.ac.uk Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers David Tomaz Nadia Guerra Julian Dyson Keith Gould Google David Tomaz Nadia Guerra Julian Dyson Keith Gould Google Scholar David Tomaz Nadia Guerra Julian Dyson Keith Gould PubMed David Tomaz Nadia Guerra Julian Dyson Keith Gould Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.