Carter, Lisa Louise; (2020) Investigating the Role of the T-Cell Receptor Using Targeted Capture and High-Throughput Sequencing. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Maintaining a diverse immune repertoire is crucial for protection against a wide range of pathogens. Until recently, it has been difficult to quantify this diversity and define the range of a repertoire in a healthy individual. The rise of massively parallel highthroughput sequencing has enabled researchers to gather more information than ever before, but many published works concentrate on describing individual T-cell and immunoglobulin chains. This report introduces a novel method for sequencing all a, b, g and d chains of the T-cell receptor and all immunoglobulin chains simultaneously, using high-throughput sequencing and targeted capture. Data was obtained through this method using two sequencing platforms, the Illumina MiSeq and the Ion Torrent Personal Genome Machine, and the analysis of data focused on the T-cell receptor using diversity measures borrowed from other scientific fields. This work demonstrated the successes and limitations of the capture technique and suggests that immune repertoire sequencing could have dramatic impact on the understanding of the immune system across a range of disease states. Therefore, a preliminary investigation was carried out into the reconstitution of the immune repertoires following paediatric haematopoeitic stem cell transplants in the treatment of acute myeloid leukaemia.

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