Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells

Sunil Kumar Saini, Tripti Tamhane, Raghavendra Anjanappa, Ankur Saikia, Sofie Ramskov, Marco Donia, Inge Marie Svane, Søren Nyboe Jakobsen, Maria Garcia-Alai, Martin Zacharias, Rob Meijers, Sebastian Springer, Sine Reker Hadrup*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the a1 and a2 helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.

Original languageEnglish
Article numbereaau9039
JournalScience Immunology
Volume4
Issue number37
Number of pages13
DOIs
Publication statusPublished - 2019

Cite this

Saini, Sunil Kumar ; Tamhane, Tripti ; Anjanappa, Raghavendra ; Saikia, Ankur ; Ramskov, Sofie ; Donia, Marco ; Svane, Inge Marie ; Jakobsen, Søren Nyboe ; Garcia-Alai, Maria ; Zacharias, Martin ; Meijers, Rob ; Springer, Sebastian ; Hadrup, Sine Reker. / Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells. In: Science Immunology. 2019 ; Vol. 4, No. 37.
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abstract = "The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the a1 and a2 helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.",
author = "Saini, {Sunil Kumar} and Tripti Tamhane and Raghavendra Anjanappa and Ankur Saikia and Sofie Ramskov and Marco Donia and Svane, {Inge Marie} and Jakobsen, {S{\o}ren Nyboe} and Maria Garcia-Alai and Martin Zacharias and Rob Meijers and Sebastian Springer and Hadrup, {Sine Reker}",
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Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells. / Saini, Sunil Kumar; Tamhane, Tripti; Anjanappa, Raghavendra; Saikia, Ankur; Ramskov, Sofie; Donia, Marco; Svane, Inge Marie; Jakobsen, Søren Nyboe; Garcia-Alai, Maria; Zacharias, Martin; Meijers, Rob; Springer, Sebastian; Hadrup, Sine Reker.

In: Science Immunology, Vol. 4, No. 37, eaau9039, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

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AU - Saini, Sunil Kumar

AU - Tamhane, Tripti

AU - Anjanappa, Raghavendra

AU - Saikia, Ankur

AU - Ramskov, Sofie

AU - Donia, Marco

AU - Svane, Inge Marie

AU - Jakobsen, Søren Nyboe

AU - Garcia-Alai, Maria

AU - Zacharias, Martin

AU - Meijers, Rob

AU - Springer, Sebastian

AU - Hadrup, Sine Reker

PY - 2019

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N2 - The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the a1 and a2 helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.

AB - The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer–based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the a1 and a2 helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2Kb can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.

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JO - Science Immunology

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SN - 2470-9468

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