title: Flanking residues are central to DO11.10 T cell hybridoma stimulation by ovalbumin 323-339.
abstract: T cell activation requires formation of a tri-molecular interaction between a major histocompatibility complex (MHC), peptide, and T cell receptor. In a common model system, the ovalbumin epitope 323-339 binds the murine class II MHC, I-A(d), in at least three distinct registers. The DO11.10 T cell recognizes the least stable of these, as determined by peptide-MHC dissociation rates. Using exogenous peptides and peptide insertions into a carrier protein in combination with IL-2 secretion assays, we show that the alternate registers do not competitively inhibit display of the active register four. In contrast, this weakly binding register is stabilized by the presence of n-terminal flanking residues active in MHC binding. The DO11.10 hybridoma is sensitive to the presence of specific wild-type residues extending to at least the P-3 peptide position. Transfer of the P-4 to P-2 flanking residues to a hen egg lysozyme epitope also presented by I-A(d) increases the activity of that epitope substantially. These results illustrate the inherent complexity in delineating the interaction of multiple registers based on traditional thermodynamic measurements and demonstrate the potential of flanking residue modification for increasing the activity of weakly bound epitopes. The latter technique represents an alternative to substitution of anchor residues within a weakly bound register, which we show can significantly decrease the activity of the epitope to a responding T cell.
authors.affiliation: Department of Chemical Engineering, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Roy
authors.forename: Dmitriy V
authors.initials: DV
authors.lastname: Zhukov
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Maynard
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D019295:Computational Biology; D000939:Epitopes / Q000737:chemistry; D006825:Hybridomas / Q000378:metabolism*; D051379:Mice; D010047:Ovalbumin / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: D000939:Epitopes; D010047:Ovalbumin