Disulfide bond engineering to trap peptides in the MHC class I binding groove

Steven M. Truscott, Lonnie Lybarger, John M. Martinko, Vesselin E. Mitaksov, David M. Kranz, Janet M. Connolly, Daved H. Fremont, Ted H. Hansen

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Immunodominant peptides in CD8 T cell responses to pathogens and tumors are not always tight binders to MHC class I molecules. Furthermore, antigenic peptides that bind weakly to the MHC can be problematic when designing vaccines to elicit CD8 T cells in vivo or for the production of MHC multimers for enumerating pathogen-specific T cells in vitro. Thus, to enhance peptide binding to MHC class I, we have engineered a disulfide bond to trap antigenic peptides into the binding groove of murine MHC class I molecules expressed as single-chain trimers or SCTs. These SCTs with disulfide traps, termed dtSCTs, oxidized properly in the endoplasmic reticulum, transited to the cell surface, and were recognized by T cells. Introducing a disulfide trap created remarkably tenacious MHC/peptide complexes because the peptide moiety of the dtSCT was not displaced by high-affinity competitor peptides, even when relatively weak binding peptides were incorporated into the dtSCT. This technology promises to be useful for DNA vaccination to elicit CD8 T cells, in vivo study of CD8 T cell development, and construction of multivalent MHC/peptide reagents for the enumeration and tracking of T cells - particularly when the antigenic peptide has relatively weak affinity for the MHC.

Original languageEnglish (US)
Pages (from-to)6280-6289
Number of pages10
JournalJournal of Immunology
Volume178
Issue number10
Publication statusPublished - May 15 2007

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ASJC Scopus subject areas

  • Immunology

Cite this

Truscott, S. M., Lybarger, L., Martinko, J. M., Mitaksov, V. E., Kranz, D. M., Connolly, J. M., ... Hansen, T. H. (2007). Disulfide bond engineering to trap peptides in the MHC class I binding groove. Journal of Immunology, 178(10), 6280-6289.