Stress-testing the relationship between T cell receptor/peptide-MHC affinity and cross-reactivity using peptide velcro
Abstract
T cell receptors (TCRs) bind to peptide-major histocompatibility complex (pMHC) with low affinity ( K d ∼ μM), which is generally assumed to facilitate cross-reactive TCR “scanning” of ligands. To understand the relationship between TCR/pMHC affinity and cross-reactivity, we sought to engineer an additional weak interaction, termed “velcro,” between the TCR and pMHC to probe the specificities of TCRs at relatively low and high affinities. This additional interaction was generated through an eight-amino acid peptide library covalently linked to the N terminus of the MHC-bound peptide. Velcro was selected through an affinity-based isolation and was subsequently shown to enhance the cognate TCR/pMHC affinity in a peptide-dependent manner by ∼10-fold. This was sufficient to convert a nonstimulatory ultra-low-affinity ligand into a stimulatory ligand. An X-ray crystallographic structure revealed how velcro interacts with the TCR. To probe TCR cross-reactivity, we screened TCRs against yeast-displayed pMHC libraries with and without velcro, and found that the peptide cross-reactivity profiles of low-affinity ( K d > 100 μM) and high-affinity ( K d ∼ μM) TCR/pMHC interactions are remarkably similar. The conservation of recognition of the TCR for pMHC across affinities reveals the nature of low-affinity ligands for which there are important biological functions andmore »
- Authors:
- Publication Date:
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1460885
- Grant/Contract Number:
- AC02-76SF00515
- Resource Type:
- Published Article
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 115 Journal Issue: 31; Journal ID: ISSN 0027-8424
- Publisher:
- Proceedings of the National Academy of Sciences
- Country of Publication:
- United States
- Language:
- English
Citation Formats
Gee, Marvin H., Sibener, Leah V., Birnbaum, Michael E., Jude, Kevin M., Yang, Xinbo, Fernandes, Ricardo A., Mendoza, Juan L., Glassman, Caleb R., and Garcia, K. Christopher. Stress-testing the relationship between T cell receptor/peptide-MHC affinity and cross-reactivity using peptide velcro. United States: N. p., 2018.
Web. doi:10.1073/pnas.1802746115.
Gee, Marvin H., Sibener, Leah V., Birnbaum, Michael E., Jude, Kevin M., Yang, Xinbo, Fernandes, Ricardo A., Mendoza, Juan L., Glassman, Caleb R., & Garcia, K. Christopher. Stress-testing the relationship between T cell receptor/peptide-MHC affinity and cross-reactivity using peptide velcro. United States. doi:10.1073/pnas.1802746115.
Gee, Marvin H., Sibener, Leah V., Birnbaum, Michael E., Jude, Kevin M., Yang, Xinbo, Fernandes, Ricardo A., Mendoza, Juan L., Glassman, Caleb R., and Garcia, K. Christopher. Wed .
"Stress-testing the relationship between T cell receptor/peptide-MHC affinity and cross-reactivity using peptide velcro". United States. doi:10.1073/pnas.1802746115.
@article{osti_1460885,
title = {Stress-testing the relationship between T cell receptor/peptide-MHC affinity and cross-reactivity using peptide velcro},
author = {Gee, Marvin H. and Sibener, Leah V. and Birnbaum, Michael E. and Jude, Kevin M. and Yang, Xinbo and Fernandes, Ricardo A. and Mendoza, Juan L. and Glassman, Caleb R. and Garcia, K. Christopher},
abstractNote = {T cell receptors (TCRs) bind to peptide-major histocompatibility complex (pMHC) with low affinity ( K d ∼ μM), which is generally assumed to facilitate cross-reactive TCR “scanning” of ligands. To understand the relationship between TCR/pMHC affinity and cross-reactivity, we sought to engineer an additional weak interaction, termed “velcro,” between the TCR and pMHC to probe the specificities of TCRs at relatively low and high affinities. This additional interaction was generated through an eight-amino acid peptide library covalently linked to the N terminus of the MHC-bound peptide. Velcro was selected through an affinity-based isolation and was subsequently shown to enhance the cognate TCR/pMHC affinity in a peptide-dependent manner by ∼10-fold. This was sufficient to convert a nonstimulatory ultra-low-affinity ligand into a stimulatory ligand. An X-ray crystallographic structure revealed how velcro interacts with the TCR. To probe TCR cross-reactivity, we screened TCRs against yeast-displayed pMHC libraries with and without velcro, and found that the peptide cross-reactivity profiles of low-affinity ( K d > 100 μM) and high-affinity ( K d ∼ μM) TCR/pMHC interactions are remarkably similar. The conservation of recognition of the TCR for pMHC across affinities reveals the nature of low-affinity ligands for which there are important biological functions and has implications for understanding the specificities of affinity-matured TCRs.},
doi = {10.1073/pnas.1802746115},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 31,
volume = 115,
place = {United States},
year = {2018},
month = {7}
}
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DOI: 10.1073/pnas.1802746115
DOI: 10.1073/pnas.1802746115
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Works referenced in this record:
T-cell receptor binding affinities and kinetics: impact on T-cell activity and specificity
journal, February 2009
- Stone, Jennifer D.; Chervin, Adam S.; Kranz, David M.
- Immunology, Vol. 126, Issue 2
An endogenous positively selecting peptide enhances mature T cell responses and becomes an autoantigen in the absence of microRNA miR-181a
journal, October 2009
- Ebert, Peter J. R.; Jiang, Shan; Xie, Jianming
- Nature Immunology, Vol. 10, Issue 11
Kinetics of T-cell receptor binding to peptide/I-Ek complexes: correlation of the dissociation rate with T-cell responsiveness.
journal, December 1994
- Matsui, K.; Boniface, J. J.; Steffner, P.
- Proceedings of the National Academy of Sciences, Vol. 91, Issue 26
Attenuated T Cell Responses to a High-Potency Ligand In Vivo
journal, September 2010
- Corse, Emily; Gottschalk, Rachel A.; Krogsgaard, Michelle
- PLoS Biology, Vol. 8, Issue 9
An endogenous peptide positively selects and augments the activation and survival of peripheral CD4+ T cells
journal, October 2009
- Lo, Wan-Lin; Felix, Nathan J.; Walters, James J.
- Nature Immunology, Vol. 10, Issue 11
Conformational changes and flexibility in T-cell receptor recognition of peptide–MHC complexes
journal, September 2008
- Armstrong, Kathryn M.; Piepenbrink, Kurt H.; Baker, Brian M.
- Biochemical Journal, Vol. 415, Issue 2
T-cell-receptor affinity and thymocyte positive selection
journal, June 1996
- Alam, S. Munir; Travers, Paul J.; Wung, Jay L.
- Nature, Vol. 381, Issue 6583
The Repertoire of T Cells Shaped by a Single MHC/Peptide Ligand
journal, February 1996
- Ignatowicz, Leszek; Kappler, John; Marrack, Philippa
- Cell, Vol. 84, Issue 4
Selecting and maintaining a diverse T-cell repertoire
journal, November 1999
- Goldrath, Ananda W.; Bevan, Michael J.
- Nature, Vol. 402, Issue 6759
Requirement for Diverse, Low-Abundance Peptides in Positive Selection of T Cells
journal, January 1999
- Barton, G. M.
- Science, Vol. 283, Issue 5398
How tcrs bind Mhcs, Peptides, and Coreceptors
journal, April 2006
- Rudolph, Markus G.; Stanfield, Robyn L.; Wilson, Ian A.
- Annual Review of Immunology, Vol. 24, Issue 1
Two-step binding mechanism for T-cell receptor recognition of peptide–MHC
journal, August 2002
- Wu, Lawren C.; Tuot, Delphine S.; Lyons, Daniel S.
- Nature, Vol. 418, Issue 6897
Lck Availability during Thymic Selection Determines the Recognition Specificity of the T Cell Repertoire
journal, September 2013
- Van Laethem, François; Tikhonova, Anastasia N.; Pobezinsky, Leonid A.
- Cell, Vol. 154, Issue 6
CD4 and CD8 binding to MHC molecules primarily acts to enhance Lck delivery
journal, September 2010
- Artyomov, M. N.; Lis, M.; Devadas, S.
- Proceedings of the National Academy of Sciences, Vol. 107, Issue 39
Insights into Cytokine–Receptor Interactions from Cytokine Engineering
journal, March 2015
- Spangler, Jamie B.; Moraga, Ignacio; Mendoza, Juan L.
- Annual Review of Immunology, Vol. 33, Issue 1
T cell positive selection by a high density, low affinity ligand
journal, April 1998
- Liu, C. -P.; Crawford, F.; Marrack, P.
- Proceedings of the National Academy of Sciences, Vol. 95, Issue 8
Class II-restricted T cell receptor engineered in vitro for higher affinity retains peptide specificity and function
journal, December 2005
- Weber, K. S.; Donermeyer, D. L.; Allen, P. M.
- Proceedings of the National Academy of Sciences, Vol. 102, Issue 52
Structural Basis of Plasticity in T Cell Receptor Recognition of a Self Peptide-MHC Antigen
journal, February 1998
- Garcia, K. C.
- Science, Vol. 279, Issue 5354
Autoreactive T cells specific for insulin B:11-23 recognize a low-affinity peptide register in human subjects with autoimmune diabetes
journal, September 2014
- Yang, J.; Chow, I. -T.; Sosinowski, T.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 41
Deficient Positive Selection of CD4 T Cells in Mice Displaying Altered Repertoires of MHC Class II–Bound Self-Peptides
journal, August 1997
- Grubin, Catherine E.; Kovats, Susan; deRoos, Paul
- Immunity, Vol. 7, Issue 2
TCR Scanning of Peptide/MHC through Complementary Matching of Receptor and Ligand Molecular Flexibility
journal, February 2014
- Hawse, William F.; De, Soumya; Greenwood, Alex I.
- The Journal of Immunology, Vol. 192, Issue 6
Structural Basis of Specificity and Cross-Reactivity in T Cell Receptors Specific for Cytochrome c –I-E k
journal, April 2011
- Newell, Evan W.; Ely, Lauren K.; Kruse, Andrew C.
- The Journal of Immunology, Vol. 186, Issue 10
T Cell Receptor Signaling Is Limited by Docking Geometry to Peptide-Major Histocompatibility Complex
journal, November 2011
- Adams, Jarrett J.; Narayanan, Samanthi; Liu, Baoyu
- Immunity, Vol. 35, Issue 5
Cohen's Conjecture, Howard's Hypothesis, and Ptashne's Ptruth: an exploration of the relationship between affinity and specificity
journal, April 2010
- Greenspan, Neil S.
- Trends in Immunology, Vol. 31, Issue 4
XDS
journal, January 2010
- Kabsch, Wolfgang
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2
TCRs with high affinity for foreign pMHC show self-reactivity
journal, December 2002
- Holler, Phillip D.; Chlewicki, Lukasz K.; Kranz, David M.
- Nature Immunology, Vol. 4, Issue 1
Reciprocal TCR-CD3 and CD4 Engagement of a Nucleating pMHCII Stabilizes a Functional Receptor Macrocomplex
journal, January 2018
- Glassman, Caleb R.; Parrish, Heather L.; Lee, Mark S.
- Cell Reports, Vol. 22, Issue 5
Why must T cells be cross-reactive?
journal, August 2012
- Sewell, Andrew K.
- Nature Reviews Immunology, Vol. 12, Issue 9
Protein binding specificity versus promiscuity
journal, February 2011
- Schreiber, Gideon; Keating, Amy E.
- Current Opinion in Structural Biology, Vol. 21, Issue 1
A very high level of crossreactivity is an essential feature of the T-cell receptor
journal, September 1998
- Mason, Don
- Immunology Today, Vol. 19, Issue 9
How the T Cell Repertoire Becomes Peptide and MHC Specific
journal, July 2005
- Huseby, Eric S.; White, Janice; Crawford, Frances
- Cell, Vol. 122, Issue 2
A Single Peptide-MHC Complex Positively Selects a Diverse and Specific CD8 T Cell Repertoire
journal, November 2009
- Wang, B.; Primeau, T. M.; Myers, N.
- Science, Vol. 326, Issue 5954
Structural interplay between germline interactions and adaptive recognition determines the bandwidth of TCR-peptide-MHC cross-reactivity
journal, November 2015
- Adams, Jarrett J.; Narayanan, Samanthi; Birnbaum, Michael E.
- Nature Immunology, Vol. 17, Issue 1
Cancer Regression and Neurological Toxicity Following Anti-MAGE-A3 TCR Gene Therapy
journal, January 2013
- Morgan, Richard A.; Chinnasamy, Nachimuthu; Abate-Daga, Daniel
- Journal of Immunotherapy, Vol. 36, Issue 2
Force-Regulated In Situ TCR–Peptide-Bound MHC Class II Kinetics Determine Functions of CD4 + T Cells
journal, September 2015
- Hong, Jinsung; Persaud, Stephen P.; Horvath, Stephen
- The Journal of Immunology, Vol. 195, Issue 8
Deletion of CD4 and CD8 Coreceptors Permits Generation of αβT Cells that Recognize Antigens Independently of the MHC
journal, November 2007
- Van Laethem, François; Sarafova, Sophia D.; Park, Jung-Hyun
- Immunity, Vol. 27, Issue 5
The Study of High-Affinity TCRs Reveals Duality in T Cell Recognition of Antigen: Specificity and Degeneracy
journal, November 2006
- Donermeyer, David L.; Weber, K. Scott; Kranz, David M.
- The Journal of Immunology, Vol. 177, Issue 10
Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma
journal, August 2013
- Linette, Gerald P.; Stadtmauer, Edward A.; Maus, Marcela V.
- Blood, Vol. 122, Issue 6
Features and development of Coot
journal, March 2010
- Emsley, P.; Lohkamp, B.; Scott, W. G.
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
Mimotopes for Alloreactive and Conventional T Cells in a Peptide–MHC Display Library
journal, April 2004
- Crawford, Frances; Huseby, Eric; White, Janice
- PLoS Biology, Vol. 2, Issue 4
Breaking the affinity ceiling for antibodies and T cell receptors
journal, September 2000
- Foote, J.; Eisen, H. N.
- Proceedings of the National Academy of Sciences, Vol. 97, Issue 20
PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010
- Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
- Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
Low affinity interaction of peptide-MHC complexes with T cell receptors
journal, December 1991
- Matsui, K.; Boniface, J.; Reay, P.
- Science, Vol. 254, Issue 5039
Direct molecular mimicry enables off-target cardiovascular toxicity by an enhanced affinity TCR designed for cancer immunotherapy
journal, January 2016
- Raman, Marine C. C.; Rizkallah, Pierre J.; Simmons, Ruth
- Scientific Reports, Vol. 6, Issue 1
Peptide-MHC heterodimers show that thymic positive selection requires a more restricted set of self-peptides than negative selection
journal, May 2010
- Juang, Jeremy; Ebert, Peter J. R.; Feng, Dan
- The Journal of Experimental Medicine, Vol. 207, Issue 6
How the TCR balances sensitivity and specificity for the recognition of self and pathogens
journal, January 2012
- Morris, Gerald P.; Allen, Paul M.
- Nature Immunology, Vol. 13, Issue 2
Thymic Selection by a Single MHC/Peptide Ligand Produces a Semidiverse Repertoire of CD4+ T Cells
journal, August 1997
- Surh, Charles D.; Lee, Dong-Sup; Fung-Leung, Wei-ping
- Immunity, Vol. 7, Issue 2
Imperfect interfaces
journal, June 1998
- Ysern, Xavier; Li, Hongmin; Mariuzza, Roy A.
- Nature Structural Biology, Vol. 5, Issue 6
Selection of a Broad Repertoire of CD4+ T Cells in H-2Ma0/0 Mice
journal, August 1997
- Tourne, Sylvie; Miyazaki, Toru; Oxenius, Annette
- Immunity, Vol. 7, Issue 2
Deconstructing the Peptide-MHC Specificity of T Cell Recognition
journal, May 2014
- Birnbaum, Michael E.; Mendoza, Juan L.; Sethi, Dhruv K.
- Cell, Vol. 157, Issue 5
Specific low-affinity recognition of major histocompatibility complex plus peptide by soluble T-cell receptor
journal, April 1992
- Weber, Susanna; Traunecker, André; Oliveri, Filippo
- Nature, Vol. 356, Issue 6372
High-affinity T cell receptor differentiates cognate peptide–MHC and altered peptide ligands with distinct kinetics and thermodynamics
journal, May 2010
- Persaud, Stephen P.; Donermeyer, David L.; Weber, K. Scott
- Molecular Immunology, Vol. 47, Issue 9
Tonic T cell signalling and T cell tolerance as opposite effects of self-recognition on dendritic cells
journal, October 2010
- Garbi, Natalio; Hämmerling, Günter J.; Probst, Hans-Christian
- Current Opinion in Immunology, Vol. 22, Issue 5
Structure of the Superantigen Staphylococcal Enterotoxin B in Complex with TCR and Peptide–MHC Demonstrates Absence of TCR–Peptide Contacts
journal, July 2014
- Rödström, Karin E. J.; Elbing, Karin; Lindkvist-Petersson, Karin
- The Journal of Immunology, Vol. 193, Issue 4