Gauging antigen recognition by human primary T-cells featuring orthotopically exchanged TCRs of choice

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Item Type:	Article
Title:	Gauging antigen recognition by human primary T-cells featuring orthotopically exchanged TCRs of choice
Creators Name:	Mühlgrabner, V., Plach, A., Holler, J., Leitner, J., Steinberger, P., Dupré, L., Göhring, J. and Huppa, J.B.
Abstract:	Understanding human T-cell antigen recognition in health and disease is becoming increasingly instrumental for monitoring T-cell responses to pathogen challenge and for the rational design of T-cell-based therapies targeting cancer, autoimmunity and organ transplant rejection. Here we showcase a quantitative imaging platform which is based on the use of planar glass-supported lipid bilayers (SLBs). The latter are functionalized with antigen (peptide-loaded HLA) as adhesion and costimulatory molecules (ICAM-1, B7-1) to serve as surrogate antigen presenting cell for antigen recognition by T-cells, which are equipped with T-cell antigen receptors (TCRs) sequenced from antigen-specific patient T-cells. We outline in detail, how the experimental use of SLBs supports recoding and analysis of synaptic antigen engagement and calcium signaling at the single cell level in response to user-defined antigen densities for quantitative comparison.
Keywords:	TCR Sequencing, T-Cell Antigen Sensitivity, CRISPR-Cas9-Mediated TCR Exchange, Advanced Live-Cell Imaging
Source:	Methods in Cell Biology
ISSN:	0091-679X
Publisher:	Elsevier
Date:	15 April 2024
Official Publication:	https://doi.org/10.1016/bs.mcb.2024.03.003

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