Photocrosslinkable pMHC monomers stain T cells specifically and cause ligand-bound TCRs to be 'preferentially' transported to the cSMAC

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Item Type:	Article
Title:	Photocrosslinkable pMHC monomers stain T cells specifically and cause ligand-bound TCRs to be 'preferentially' transported to the cSMAC
Creators Name:	Xie, J., Huppa, J.B., Newell, E.W., Huang, J., Ebert, P.J.R., Li, Q.J. and Davis, M.M.
Abstract:	The binding of T cell antigen receptors (TCRs) to specific complexes of peptide and major histocompatibility complex (pMHC) is typically of very low affinity, which necessitates the use of multimeric pMHC complexes to label T lymphocytes stably. We report here the development of pMHC complexes able to be crosslinked by ultraviolet irradiation; even as monomers, these efficiently and specifically stained cognate T cells. We also used this reagent to probe T cell activation and found that a covalently bound pMHC was more stimulatory than an agonist pMHC on lipid bilayers. This finding suggested that serial engagement of TCRs is dispensable for activation when a substantial fraction of TCRs are stably engaged. Finally, pMHC-bound TCRs were 'preferentially' transported into the central supramolecular activation cluster after activation, which suggested that ligand engagement enabled linkage of the TCR and its associated CD3 signaling molecules to the cytoskeleton.
Keywords:	CD3 Complex, Cultured Cells, Coloring Agents, Cross-Linking Reagents, Cytoskeleton, Lymphocyte Activation, Major Histocompatibility Complex, Transgenic Mice, T-Cell Antigen Receptors, Signal Transduction, T-Lymphocytes, T-Lymphocytes / Immunology, Animals, Mice
Source:	Nature Immunology
ISSN:	1529-2908
Publisher:	Nature Publishing Group
Volume:	13
Number:	7
Page Range:	674-80
Date:	July 2012
Official Publication:	https://doi.org/10.1038/ni.2344
PubMed:	View item in PubMed

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