Massively parallel interrogation and mining of natively paired human TCRαβ repertoires
- 16 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Biotechnology
- Vol. 38 (5), 609-619
- https://doi.org/10.1038/s41587-020-0438-y
Abstract
T cells engineered to express antigen-specific T cell receptors (TCRs) are potent therapies for viral infections and cancer. However, efficient identification of clinical candidate TCRs is complicated by the size and complexity of T cell repertoires and the challenges of working with primary T cells. Here we present a high-throughput method to identify TCRs with high functional avidity from diverse human T cell repertoires. The approach used massively parallel microfluidics to generate libraries of natively paired, full-length TCRαβ clones, from millions of primary T cells, which were then expressed in Jurkat cells. The TCRαβ–Jurkat libraries enabled repeated screening and panning for antigen-reactive TCRs using peptide major histocompatibility complex binding and cellular activation. We captured more than 2.9 million natively paired TCRαβ clonotypes from six healthy human donors and identified rare (<0.001% frequency) viral-antigen-reactive TCRs. We also mined a tumor-infiltrating lymphocyte sample from a patient with melanoma and identified several tumor-specific TCRs, which, after expression in primary T cells, led to tumor cell killing.Funding Information
- U.S. Department of Health & Human Services | NIH | National Cancer Institute (R43CA232942)
- RCUK | Engineering and Physical Sciences Research Council (EP/L014904/1)
- U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (R43AI120313-01)
This publication has 67 references indexed in Scilit:
- Durable Complete Responses in Heavily Pretreated Patients with Metastatic Melanoma Using T-Cell Transfer ImmunotherapyClinical Cancer Research, 2011
- Search and clustering orders of magnitude faster than BLASTBioinformatics, 2010
- Enhanced receptor expression and in vitro effector function of a murine-human hybrid MART-1-reactive T cell receptor following a rapid expansionCancer Immunology, Immunotherapy, 2010
- Germ Line-governed Recognition of a Cancer Epitope by an Immunodominant Human T-cell ReceptorJournal of Biological Chemistry, 2009
- IMGT(R), the international ImMunoGeneTics information system(R)Nucleic Acids Research, 2008
- Expression of heterologous genes in oncolytic adenoviruses using picornaviral 2A sequences that trigger ribosome skippingJournal of General Virology, 2008
- Influence of Human CD8 on Antigen Recognition by T-Cell Receptor–Transduced CellsCancer Research, 2006
- Deconstructing the Form and Function of the TCR/CD3 ComplexImmunity, 2006
- A biased Vα24+ T-cell repertoire leads to circulating NKT-cell defects in a multiple sclerosis patient at the onset of his diseaseImmunology Letters, 2003
- CD161 (NKR-P1A) Costimulation of CD1d-dependent Activation of Human T Cells Expressing Invariant Vα24JαQ T Cell Receptor α ChainsThe Journal of Experimental Medicine, 1998