Using CombiCells, a platform enabling titration and combinatorial display of cell surface ligands, to investigate the sensitivity and costimulatory requirements of TCRs and CARs

Patel, A.; Andre, V.; Bustamante Eguiguren, S.; Barton, M. I.; Denham, E. M.; Pettmann, J.; Morch, A. M.; Kutuzov, M.; Siller-Farfan, J. A.; Dustin, M. L.; van der Merwe, P. A.; Dushek, O.

doi:10.1101/2023.06.15.545075

bioRxiv · 10.1101/2023.06.15.545075

Using CombiCells, a platform enabling titration and combinatorial display of cell surface ligands, to investigate the sensitivity and costimulatory requirements of TCRs and CARs

Patel, A.·Andre, V.·Bustamante Eguiguren, S.·Barton, M. I.·Denham, E. M.·Pettmann, J.·Morch, A. M.·Kutuzov, M.·Siller-Farfan, J. A.·Dustin, M. L.·van der Merwe, P. A.·Dushek, O.

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Abstract

Understanding how cellular decisions by receptor/ligand interactions at cell/cell interface has been challenging because it is difficult to independently vary the surface density of multiple ligands. Here, we exploit the SpyCatcher/SpyTag split-protein system for rapid combinatorial display of native ligands on cells (Combicells). We use this platform to assess T cell antigen sensitivity and the impact of T cell co-stimulation/co-inhibition receptors. The TCR displayed much greater sensitivity to pMHC than CARs and BiTES did to CD19. While TCR sensitivity was greatly enhanced by CD2 ligand, CAR sensitivity to CD19 was primarily but more modestly enhanced by LFA-1 ligand. Lastly, we show that the PD-1/ligand engagement inhibited T cell activation triggered solely by TCR/pMHC interactions, as well as the amplified activation induced by CD2 and CD28 co-stimulation. The ability to easily produce cells with different concentrations and combinations of ligands should accelerate the study of receptor/ligand interactions at cell/cell interfaces. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=177 SRC="FIGDIR/small/545075v2_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@d1ec95org.highwire.dtl.DTLVardef@33c0forg.highwire.dtl.DTLVardef@5bfed7org.highwire.dtl.DTLVardef@1b80ee2_HPS_FORMAT_FIGEXP M_FIG C_FIG One sentence summaryUsing CombiCells, a platform for the combinatorial display of cell surface ligands, to compare T cell antigen sensitivity mediated by TCRs, CARs, and BiTEs and its dependence on co-stimulation/co-inhibition receptor ligands

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Patel, A., Andre, V., Bustamante Eguiguren, S., Barton, M. I., Denham, E. M., Pettmann, J., Morch, A. M., Kutuzov, M., Siller-Farfan, J. A., Dustin, M. L., van der Merwe, P. A., Dushek, O.. 2023-06-17. Using CombiCells, a platform enabling titration and combinatorial display of cell surface ligands, to investigate the sensitivity and costimulatory requirements of TCRs and CARs. https://doi.org/10.1101/2023.06.15.545075

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