Cell activation-based screening of natively paired human T cell receptor repertoires
Adoptive immune therapies based on the transfer of antigen-specific T cells have been used successfully to treat various cancers and viral infections, but improved techniques are needed to identify optimally protective human T cell receptors (TCRs). Here we present a high-throughput approach to the...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Springer Science and Business Media LLC
2025
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| Online Access: | https://hdl.handle.net/1721.1/158239 |
| _version_ | 1824458253206028288 |
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| author | Fahad, Ahmed S Chung, Cheng Yu López Acevedo, Sheila N Boyle, Nicoleen Madan, Bharat Gutiérrez-González, Matías F Matus-Nicodemos, Rodrigo Laflin, Amy D Ladi, Rukmini R Zhou, John Wolfe, Jacy Llewellyn-Lacey, Sian Koup, Richard A Douek, Daniel C Balfour, Henry H Price, David A DeKosky, Brandon J |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Fahad, Ahmed S Chung, Cheng Yu López Acevedo, Sheila N Boyle, Nicoleen Madan, Bharat Gutiérrez-González, Matías F Matus-Nicodemos, Rodrigo Laflin, Amy D Ladi, Rukmini R Zhou, John Wolfe, Jacy Llewellyn-Lacey, Sian Koup, Richard A Douek, Daniel C Balfour, Henry H Price, David A DeKosky, Brandon J |
| author_sort | Fahad, Ahmed S |
| collection | MIT |
| description | Adoptive immune therapies based on the transfer of antigen-specific T cells have been used successfully to treat various cancers and viral infections, but improved techniques are needed to identify optimally protective human T cell receptors (TCRs). Here we present a high-throughput approach to the identification of natively paired human TCRα and TCRβ (TCRα:β) genes encoding heterodimeric TCRs that recognize specific peptide antigens bound to major histocompatibility complex molecules (pMHCs). We first captured and cloned TCRα:β genes from individual cells, ensuring fidelity using a suppression PCR. We then screened TCRα:β libraries expressed in an immortalized cell line using peptide-pulsed antigen-presenting cells and sequenced activated clones to identify the cognate TCRs. Our results validated an experimental pipeline that allows large-scale repertoire datasets to be annotated with functional specificity information, facilitating the discovery of therapeutically relevant TCRs. |
| first_indexed | 2025-02-19T04:22:57Z |
| format | Article |
| id | mit-1721.1/158239 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2025-02-19T04:22:57Z |
| publishDate | 2025 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1582392025-02-18T23:34:52Z Cell activation-based screening of natively paired human T cell receptor repertoires Fahad, Ahmed S Chung, Cheng Yu López Acevedo, Sheila N Boyle, Nicoleen Madan, Bharat Gutiérrez-González, Matías F Matus-Nicodemos, Rodrigo Laflin, Amy D Ladi, Rukmini R Zhou, John Wolfe, Jacy Llewellyn-Lacey, Sian Koup, Richard A Douek, Daniel C Balfour, Henry H Price, David A DeKosky, Brandon J Massachusetts Institute of Technology. Department of Chemical Engineering Ragon Institute of MGH, MIT and Harvard Adoptive immune therapies based on the transfer of antigen-specific T cells have been used successfully to treat various cancers and viral infections, but improved techniques are needed to identify optimally protective human T cell receptors (TCRs). Here we present a high-throughput approach to the identification of natively paired human TCRα and TCRβ (TCRα:β) genes encoding heterodimeric TCRs that recognize specific peptide antigens bound to major histocompatibility complex molecules (pMHCs). We first captured and cloned TCRα:β genes from individual cells, ensuring fidelity using a suppression PCR. We then screened TCRα:β libraries expressed in an immortalized cell line using peptide-pulsed antigen-presenting cells and sequenced activated clones to identify the cognate TCRs. Our results validated an experimental pipeline that allows large-scale repertoire datasets to be annotated with functional specificity information, facilitating the discovery of therapeutically relevant TCRs. 2025-02-18T23:34:51Z 2025-02-18T23:34:51Z 2023 2025-02-18T23:12:25Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/158239 Fahad, A.S., Chung, C.Y., López Acevedo, S.N. et al. Cell activation-based screening of natively paired human T cell receptor repertoires. Sci Rep 13, 8011 (2023). en 10.1038/s41598-023-31858-4 Scientific Reports Creative Commons Attribution https://creativecommons.org/licenses/by/4.0/ application/pdf Springer Science and Business Media LLC Springer Science and Business Media LLC |
| spellingShingle | Fahad, Ahmed S Chung, Cheng Yu López Acevedo, Sheila N Boyle, Nicoleen Madan, Bharat Gutiérrez-González, Matías F Matus-Nicodemos, Rodrigo Laflin, Amy D Ladi, Rukmini R Zhou, John Wolfe, Jacy Llewellyn-Lacey, Sian Koup, Richard A Douek, Daniel C Balfour, Henry H Price, David A DeKosky, Brandon J Cell activation-based screening of natively paired human T cell receptor repertoires |
| title | Cell activation-based screening of natively paired human T cell receptor repertoires |
| title_full | Cell activation-based screening of natively paired human T cell receptor repertoires |
| title_fullStr | Cell activation-based screening of natively paired human T cell receptor repertoires |
| title_full_unstemmed | Cell activation-based screening of natively paired human T cell receptor repertoires |
| title_short | Cell activation-based screening of natively paired human T cell receptor repertoires |
| title_sort | cell activation based screening of natively paired human t cell receptor repertoires |
| url | https://hdl.handle.net/1721.1/158239 |
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