Abstract
The binding of oligomeric peptide-MHC (pMHC) complexes to cell surface TCR can be considered to approximate TCR-pMHCinteractions at cell-cell interfaces. In this study, we analyzed the equilibrium binding of streptavidin-based pMHC oligomers(tetramers) and their dissociation kinetics from CD8 pos T cells from 2C-TCR transgenic mice and from T cell hybridomas thatexpressed the 2C TCR or a high-affinity mutant (m33) of this TCR. Our results show that the tetramers did not come close tosaturating cell-surface TCR (binding only 10-30% of cell-surface receptors), as is generally assumed in deriving affinity values(K D), in part because of dissociative losses from tetramer-stained cells. Guided by a kinetic model, the oligomer dissociation rateand equilibrium constants were seen to depend not only on monovalent association and dissociation rates (k off and k on), but also ona multivalent association rate (μ) and TCR cell-surface density. Our results suggest that dissociation rates could account for therecently described surprisingly high frequency of tetramer-negative, functionally competent T cells in some T cell responses.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6281-6290 |
| Number of pages | 10 |
| Journal | Journal of Immunology |
| Volume | 187 |
| Issue number | 12 |
| DOIs | |
| State | Published - Dec 15 2011 |
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ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Interaction of streptavidin-based peptide-MHC oligomers (tetramers) with cell-surface TCRs. / Stone, Jennifer D.; Artyomov, Maxim N.; Chervin, Adam S.; Chakraborty, Arup K.; Eisen, Herman N.; Kranz, David M.
In: Journal of Immunology, Vol. 187, No. 12, 15.12.2011, p. 6281-6290.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Interaction of streptavidin-based peptide-MHC oligomers (tetramers) with cell-surface TCRs
AU - Stone, Jennifer D.
AU - Artyomov, Maxim N.
AU - Chervin, Adam S.
AU - Chakraborty, Arup K.
AU - Eisen, Herman N.
AU - Kranz, David M
PY - 2011/12/15
Y1 - 2011/12/15
N2 - The binding of oligomeric peptide-MHC (pMHC) complexes to cell surface TCR can be considered to approximate TCR-pMHCinteractions at cell-cell interfaces. In this study, we analyzed the equilibrium binding of streptavidin-based pMHC oligomers(tetramers) and their dissociation kinetics from CD8 pos T cells from 2C-TCR transgenic mice and from T cell hybridomas thatexpressed the 2C TCR or a high-affinity mutant (m33) of this TCR. Our results show that the tetramers did not come close tosaturating cell-surface TCR (binding only 10-30% of cell-surface receptors), as is generally assumed in deriving affinity values(K D), in part because of dissociative losses from tetramer-stained cells. Guided by a kinetic model, the oligomer dissociation rateand equilibrium constants were seen to depend not only on monovalent association and dissociation rates (k off and k on), but also ona multivalent association rate (μ) and TCR cell-surface density. Our results suggest that dissociation rates could account for therecently described surprisingly high frequency of tetramer-negative, functionally competent T cells in some T cell responses.
AB - The binding of oligomeric peptide-MHC (pMHC) complexes to cell surface TCR can be considered to approximate TCR-pMHCinteractions at cell-cell interfaces. In this study, we analyzed the equilibrium binding of streptavidin-based pMHC oligomers(tetramers) and their dissociation kinetics from CD8 pos T cells from 2C-TCR transgenic mice and from T cell hybridomas thatexpressed the 2C TCR or a high-affinity mutant (m33) of this TCR. Our results show that the tetramers did not come close tosaturating cell-surface TCR (binding only 10-30% of cell-surface receptors), as is generally assumed in deriving affinity values(K D), in part because of dissociative losses from tetramer-stained cells. Guided by a kinetic model, the oligomer dissociation rateand equilibrium constants were seen to depend not only on monovalent association and dissociation rates (k off and k on), but also ona multivalent association rate (μ) and TCR cell-surface density. Our results suggest that dissociation rates could account for therecently described surprisingly high frequency of tetramer-negative, functionally competent T cells in some T cell responses.
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U2 - 10.4049/jimmunol.1101734
DO - 10.4049/jimmunol.1101734
M3 - Article
VL - 187
SP - 6281
EP - 6290
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 12
ER -