Overview of methodologies for T-cell receptor repertoire analysis

Abstract

BackgroundThe T-cell receptor (TCR), located on the surface of T cells, is responsible for the recognition of the antigen-major histocompatibility complex, leading to the initiation of an inflammatory response. Analysing the TCR repertoire may help to gain a better understanding of the immune system features and of the aetiology and progression of diseases, in particular those with unknown antigenic triggers. The extreme diversity of the TCR repertoire represents a major analytical challenge; this has led to the development of specialized methods which aim to characterize the TCR repertoire in-depth. Currently, next generation sequencing based technologies are most widely employed for the high-throughput analysis of the immune cell repertoire.ResultsHere, we report on the latest methodological advancements in the field by describing and comparing the available tools; from the choice of the starting material and library preparation method, to the sequencing technologies and data analysis.Finally, we provide a practical example and our own experience by reporting some exemplary results from a small internal benchmark study, where current approaches from the literature and the market are employed and compared.ConclusionsSeveral valid methods for clonotype identification and TCR repertoire analysis exist, however, a gold standard method for the field has not yet been identified. Depending on the purpose of the scientific study, some approaches may be more suitable than others. Finally, due to possible method specific biases, scientists must be careful when comparing results obtained using different methods.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0379-9) contains supplementary material, which is available to authorized users.

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“…16 BCR sequencing also measures somatic hypermutation. 16 BCR sequencing also measures somatic hypermutation.…”

“…16 In targeted protocols, the antigen receptor gene must be amplified to ensure sequence detection in a high proportion of cells. 16 In targeted protocols, the antigen receptor gene must be amplified to ensure sequence detection in a high proportion of cells.…”

“…16 In addition, identification of BCR gene segments and rate of somatic hypermutation depends on alignment to a germline database. 16 In addition, identification of BCR gene segments and rate of somatic hypermutation depends on alignment to a germline database.…”

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