Optimisation of T cell receptor antigen recognition for targeting disease

Doctoral thesis English OPEN
Lissina, Anna
  • Subject: QR180
    mesheuropmc: chemical and pharmacologic phenomena

This thesis is a collection of some of the studies I have undertaken over the last 3.5 years while working as a Research Assistant in the T cell modulation group at the Cardiff University School of Medicine. The work contained within is linked by the common theme of optimising interactions between peptide-Major Histocompatibility (pMHC) molecules, the T cell receptor (TCR), and/or coreceptor that engages these ligands. The work of the T cell modulation group is heavily focused on translational medicine. This aspect of biomedicine is also strongly encouraged by my funding body, the Wellcome Trust. My focus on translational aspects of interactions with pMHC ligands took my work in several different directions. Initially, I examined ways of improving interactions with pMHC that could be used to ameliorate the detection of antigen-specific T cells by flow cytometry. My studies have improved this technology to a point where I can now reliably claim to be able to stain the majority of relevant T cells with their cognate multimeric antigen. The approaches I helped pioneer are now in use all over the world. This work is reported in Chapters 3 and 5, and has resulted in two published primary data papers. A third paper that examines pMHC multimer valency (described in Chapter 4) is in preparation. In addition to the above-mentioned work aimed at improving T cell-related diagnostics using pMHC multimers, I also explored potential ways of improving TCR/pMHC interactions for therapeutic approaches. Specifically, I was interested in exploring whether TCRs displaying enhanced affinities for antigen would be useful in the clinic. These studies necessitated that we establish optimal TCR gene transfer protocols in Cardiff. I took the lead on these optimisation studies (Chapter 6). With the TCR gene transfer technology optimised, I was able to investigate whether increasing functional avidity of TCR-redirected T cells could be achieved by removing defined N-glycosylation sites within the TCR constant domain. This work was based on my observation that the desialylation of T cells improved the surface engagement of pMHC multimers and the recognition of cognate antigen when displayed naturally on a target cell surface. These studies were taken forward in Chapter 7. As part of my work with affinity enhanced TCRs, I was fortunate to test a novel set of TCR-based soluble therapeutic reagents comprising affinity-enhanced TCRs (Chapter 8). The enhanced TCRs were generated by phage display and directed evolution using techniques that were pioneered by my T cell modulation group colleague Jonathan Boulter while working at Avidex Ltd.
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