UBC Theses and Dissertations
High-throughput pairing of antigen receptor chains Mewis, Georgia
The specificity of antigen recognition by a T cell or B cell is determined by its unique T cell receptor (TCR) or B cell receptor (BCR), each consisting of two, paired polypeptide chains (alpha and beta, or heavy and light, respectively). An immense diversity of receptors is created during T cell and B cell development through a process of gene recombination. Previously, this diversity has been studied by extracting RNA from large numbers of cells, amplifying the alpha and beta chain (or heavy and light chain) transcripts, and then deep sequencing. However, through this process, information on correct chain pairing is lost. In this thesis, I present a high-throughput approach for maintaining paired-chain information in next-generation sequencing libraries. Briefly, a bulk cell population is divided into a number of sub-populations, and TCR or BCR transcripts are independently amplified; chains are considered paired if they co-occur in more sub-populations than expected by random chance. Fundamental to this approach is a reliable, sensitive library preparation chemistry in which a sub-population specific index can be incorporated. Such a chemistry was validated on primary human CD8⁺ T cells. This approach for antigen receptor chain pairing will enable in-depth studies of immune dynamics, tracking of disease progression, and personalized immunotherapeutics.
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Attribution-NonCommercial-NoDerivs 2.5 Canada