A cloning and expression system to probe T-cell receptor specificity and assess functional avidity to neoantigens.

Autor:	Hu Z; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Anandappa AJ; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA., Sun J; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Kim J; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Leet DE; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA., Bozym DJ; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA., Chen C; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Williams L; Broad Institute of MIT and Harvard, Cambridge, MA., Shukla SA; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Broad Institute of MIT and Harvard, Cambridge, MA.; Translational Immunogenomics Laboratory, Dana-Farber Cancer Institute, Boston, MA., Zhang W; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Tabbaa D; Broad Institute of MIT and Harvard, Cambridge, MA., Steelman S; Broad Institute of MIT and Harvard, Cambridge, MA., Olive O; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Livak KJ; Translational Immunogenomics Laboratory, Dana-Farber Cancer Institute, Boston, MA., Kishi H; Department of Immunology, University of Toyama, Toyama, Japan., Muraguchi A; Department of Immunology, University of Toyama, Toyama, Japan., Guleria I; Department of Pathology, Brigham and Women's Hospital, Boston, MA., Stevens J; Department of Pathology, Brigham and Women's Hospital, Boston, MA., Lane WJ; Harvard Medical School, Boston, MA.; Department of Pathology, Brigham and Women's Hospital, Boston, MA., Burkhardt UE; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA., Fritsch EF; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Broad Institute of MIT and Harvard, Cambridge, MA., Neuberg D; Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute, Boston, MA., Ott PA; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA.; Department of Medicine, Brigham and Women's Hospital, Boston, MA; and., Keskin DB; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA.; Broad Institute of MIT and Harvard, Cambridge, MA.; Department of Medicine, Brigham and Women's Hospital, Boston, MA; and., Hacohen N; Harvard Medical School, Boston, MA.; Broad Institute of MIT and Harvard, Cambridge, MA.; Massachusetts General Hospital, Boston, MA., Wu CJ; Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA.; Harvard Medical School, Boston, MA.; Broad Institute of MIT and Harvard, Cambridge, MA.; Department of Medicine, Brigham and Women's Hospital, Boston, MA; and.
Jazyk:	angličtina
Zdroj:	Blood [Blood] 2018 Nov 01; Vol. 132 (18), pp. 1911-1921. Date of Electronic Publication: 2018 Aug 27.
DOI:	10.1182/blood-2018-04-843763
Abstrakt:	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- MGA, and use this approach to identify the mut- MGA -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. (© 2018 by The American Society of Hematology.)
Databáze:	MEDLINE
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