Deep sequencing of T-Cell and B-Cell receptors with next-generation DNA sequencers

Abstract

Both T and B lymphocytes work as key effectors of adaptive immunity, recognizing a broad range of antigens with their specialized receptors (B-cell receptor in B lymphocytes, T-cell receptor in T lymphocytes). The unique rearrangement of variable (V), diversity (D), and joining (J) gene segments and the somatic hypermutations (SHM) of BCR and TCR create immense immune repertoires. Approaches such as flow cytometry, CDR3 spectratyping, or capillarybased sequencing are unable to comprehensively characterize extremely diverse immune repertoires, whereas recent advances in next-generation sequencing (NGS) enable us to examine such complicated repertoires and unravel the complexity of the immune repertoires. Application of NGS to TCR and BCR analysis has great potential to improve the monitoring of lymphoid malignancies and the assessing of immune reconstitution after hematopoietic stem cell transplantation, to better characterize immune system dysfunction in various disease conditions, including autoimmune diseases and food and drug allergies, and to determine the immune responses in various treatments (immunotherapy, radiation therapy, and chemotherapy). Here we review the role of NGS in analyzing these immune repertoires and discuss how to optimize a protocol for an unbiased preparation of TCR and BCR libraries for NGS immune repertoire analysis.