A cloning and expression system to probe T-cell receptor specificity and assess functional avidity to neoantigens.
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Abstract | Recent studies have highlighted the promise of targeting tumor neoantigens to generate potent antitumor immune responses and provide strong motivation for improving our understanding of antigen-T-cell receptor (TCR) interactions. Advances in single-cell sequencing technologies have opened the door for detailed investigation of the TCR repertoire, providing paired information from TCRα and TCRβ, which together determine specificity. However, a need remains for efficient methods to assess the specificity of discovered TCRs. We developed a streamlined approach for matching TCR sequences with cognate antigen through on-demand cloning and expression of TCRs and screening against candidate antigens. Here, we first demonstrate the system's capacity to identify viral-antigen-specific TCRs and compare the functional avidity of TCRs specific for a given antigen target. We then apply this system to identify neoantigen-specific TCR sequences from patients with melanoma treated with personalized neoantigen vaccines and characterize functional avidity of neoantigen-specific TCRs. Furthermore, we use a neoantigen-prediction pipeline to show that an insertion-deletion mutation in a putative chronic lymphocytic leukemia (CLL) driver gives rise to an immunogenic neoantigen mut- and use this approach to identify the mut--specific TCR sequence. This approach provides a means to identify and express TCRs, and then rapidly assess antigen specificity and functional avidity of a reconstructed TCR, which can be applied for monitoring antigen-specific T-cell responses, and potentially for guiding the design of effective T-cell-based immunotherapies. |
Year of Publication | 2018
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Journal | Blood
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Volume | 132
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Issue | 18
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Pages | 1911-1921
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Date Published | 2018 11 01
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ISSN | 1528-0020
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DOI | 10.1182/blood-2018-04-843763
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PubMed ID | 30150207
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PubMed Central ID | PMC6213317
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Grant list | HHMI / Howard Hughes Medical Institute / United States
R50 CA211482 / CA / NCI NIH HHS / United States
R21 CA220147 / CA / NCI NIH HHS / United States
R01 CA155010 / CA / NCI NIH HHS / United States
T32 CA207021 / CA / NCI NIH HHS / United States
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