Interaction of streptavidin-based peptide-MHC oligomers (tetramers) with cell-surface TCRs

Jennifer D. Stone, Maxim N. Artyomov, Adam S. Chervin, Arup K. Chakraborty, Herman N. Eisen, David M. Kranz

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

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 languageEnglish
Pages (from-to)6281-6290
Number of pages10
JournalJournal of Immunology
Volume187
Issue number12
DOIs
StatePublished - 15 Dec 2011
Externally publishedYes

Funding

FundersFunder number
NIH Office of the DirectorDP1OD001022
National Cancer InstituteP01CA097296
National Institute of General Medical SciencesR01GM055767
National Institute of Allergy and Infectious DiseasesP01AI071195

    Fingerprint

    Dive into the research topics of 'Interaction of streptavidin-based peptide-MHC oligomers (tetramers) with cell-surface TCRs'. Together they form a unique fingerprint.

    Cite this