High-throughput pairing of T cell receptor α and β sequences

• Published in: Science translational medicine 2015-08, Vol.7 (301), p.301ra131-301ra131
• Main Authors: Howie, Bryan, Sherwood, Anna M, Berkebile, Ashley D, Berka, Jan, Emerson, Ryan O, Williamson, David W, Kirsch, Ilan, Vignali, Marissa, Rieder, Mark J, Carlson, Christopher S, Robins, Harlan S
• Format: Article
• Language: English
• Subjects: CD8-Positive T-Lymphocytes - metabolism; Humans; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - metabolism; Receptors, Antigen, T-Cell, alpha-beta - genetics; Receptors, Antigen, T-Cell, alpha-beta - metabolism
• ISSN: 1946-6234; 1946-6242
• DOI: 10.1126/scitranslmed.aac5624

The T cell receptor (TCR) protein is a heterodimer composed of an α chain and a β chain. TCR genes undergo somatic DNA rearrangements to generate the diversity of T cell binding specificities needed for effective immunity. Recently, high-throughput immunosequencing methods have been developed to profile the TCR α (TCRA) and TCR β (TCRB) repertoires. However, these methods cannot determine which TCRA and TCRB chains combine to form a specific TCR, which is essential for many functional and therapeutic applications. We describe and validate a method called pairSEQ, which can leverage the diversity of TCR sequences to accurately pair hundreds of thousands of TCRA and TCRB sequences in a single experiment. Our TCR pairing method uses standard laboratory consumables and equipment without the need for single-cell technologies. We show that pairSEQ can be applied to T cells from both blood and solid tissues, such as tumors.