Interaction of Streptavidin-Based Peptide-MHC Oligomers (Tetramers) with Cell-Surface T Cell Receptors
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Chakraborty_Interaction of Streptavidin.pdf
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Author(s)
Stone, Jennifer D.
Artyomov, Maxim N.
Chervin, Adam S.
Eisen, Herman N.
Kranz, David
Chakraborty, Arup K
Date Issued
November 2011
Journal
Journal of Immunology
Publisher
American Association of Immunologists
Citation
Stone, J. D. et al. “Interaction of Streptavidin-Based Peptide-MHC Oligomers (Tetramers) with Cell-Surface TCRs.” The Journal of Immunology 187.12 (2011): 6281–6290.
Version
Author's final manuscript
Abstract
he binding of oligomeric peptide–MHC (pMHC) complexes to cell surface TCR can be considered to approximate TCR–pMHC interactions at cell-cell interfaces. In this study, we analyzed the equilibrium binding of streptavidin-based pMHC oligomers (tetramers) and their dissociation kinetics from CD8[superscript pos] T cells from 2C-TCR transgenic mice and from T cell hybridomas that expressed the 2C TCR or a high-affinity mutant (m33) of this TCR. Our results show that the tetramers did not come close to saturating cell-surface TCR (binding only 10–30% of cell-surface receptors), as is generally assumed in deriving affinity values (K[subscript D]), in part because of dissociative losses from tetramer-stained cells. Guided by a kinetic model, the oligomer dissociation rate and equilibrium constants were seen to depend not only on monovalent association and dissociation rates (k[subscript off] and k[subscript on]), but also on a multivalent association rate (μ) and TCR cell-surface density. Our results suggest that dissociation rates could account for the recently described surprisingly high frequency of tetramer-negative, functionally competent T cells in some T cell responses.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Massachusetts Institute of Technology. Computer Science and Artificial Intelligence Laboratory
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemical Engineering
Massachusetts Institute of Technology. Department of Chemistry
Ragon Institute of MGH, MIT and Harvard
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.4049/jimmunol.1101734
