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

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
DOIs
StatePublished - May 15 2007

Fingerprint

Disulfides
Peptides
T-Lymphocytes
Endoplasmic Reticulum
Vaccination
Vaccines
Technology
DNA

ASJC Scopus subject areas

  • Immunology and Allergy
  • 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. https://doi.org/10.4049/jimmunol.178.10.6280

Disulfide bond engineering to trap peptides in the MHC class I binding groove. / Truscott, Steven M.; Lybarger, Lonnie; Martinko, John M.; Mitaksov, Vesselin E.; Kranz, David M; Connolly, Janet M.; Fremont, Daved H.; Hansen, Ted H.

In: Journal of Immunology, Vol. 178, No. 10, 15.05.2007, p. 6280-6289.

Research output: Contribution to journalArticle

Truscott, SM, Lybarger, L, Martinko, JM, Mitaksov, VE, Kranz, DM, Connolly, JM, Fremont, DH & Hansen, TH 2007, 'Disulfide bond engineering to trap peptides in the MHC class I binding groove', Journal of Immunology, vol. 178, no. 10, pp. 6280-6289. https://doi.org/10.4049/jimmunol.178.10.6280
Truscott SM, Lybarger L, Martinko JM, Mitaksov VE, Kranz DM, Connolly JM et al. Disulfide bond engineering to trap peptides in the MHC class I binding groove. Journal of Immunology. 2007 May 15;178(10):6280-6289. https://doi.org/10.4049/jimmunol.178.10.6280
Truscott, Steven M. ; Lybarger, Lonnie ; Martinko, John M. ; Mitaksov, Vesselin E. ; Kranz, David M ; Connolly, Janet M. ; Fremont, Daved H. ; Hansen, Ted H. / Disulfide bond engineering to trap peptides in the MHC class I binding groove. In: Journal of Immunology. 2007 ; Vol. 178, No. 10. pp. 6280-6289.
@article{55708ff715fc41dd8c99ece0306726a8,
title = "Disulfide bond engineering to trap peptides in the MHC class I binding groove",
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.",
author = "Truscott, {Steven M.} and Lonnie Lybarger and Martinko, {John M.} and Mitaksov, {Vesselin E.} and Kranz, {David M} and Connolly, {Janet M.} and Fremont, {Daved H.} and Hansen, {Ted H.}",
year = "2007",
month = "5",
day = "15",
doi = "10.4049/jimmunol.178.10.6280",
language = "English (US)",
volume = "178",
pages = "6280--6289",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "10",

}

TY - JOUR

T1 - Disulfide bond engineering to trap peptides in the MHC class I binding groove

AU - Truscott, Steven M.

AU - Lybarger, Lonnie

AU - Martinko, John M.

AU - Mitaksov, Vesselin E.

AU - Kranz, David M

AU - Connolly, Janet M.

AU - Fremont, Daved H.

AU - Hansen, Ted H.

PY - 2007/5/15

Y1 - 2007/5/15

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=34248212778&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34248212778&partnerID=8YFLogxK

U2 - 10.4049/jimmunol.178.10.6280

DO - 10.4049/jimmunol.178.10.6280

M3 - Article

C2 - 17475856

AN - SCOPUS:34248212778

VL - 178

SP - 6280

EP - 6289

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 10

ER -