title: Interaction of streptavidin-based peptide-MHC oligomers (tetramers) with cell-surface TCRs.
abstract: The 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(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(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(off) and k(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.
authors.affiliation: Department of Biochemistry, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA. jstone@illinois.edu
authors.forename: Jennifer D
authors.initials: JD
authors.lastname: Stone
authors.forename: Maxim N
authors.initials: MN
authors.lastname: Artyomov
authors.forename: Adam S
authors.initials: AS
authors.lastname: Chervin
authors.forename: Arup K
authors.initials: AK
authors.lastname: Chakraborty
authors.forename: Herman N
authors.initials: HN
authors.lastname: Eisen
authors.forename: David M
authors.initials: DM
authors.lastname: Kranz
mesh_terms: D000818:Animals; D006825:Hybridomas; D008285:Major Histocompatibility Complex / Q000235:genetics / Q000276:immunology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D018448:Models, Immunological; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000378:metabolism; D019809:Streptavidin / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D046912:Multiprotein Complexes; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta; D057127:Single-Chain Antibodies; D019809:Streptavidin
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