Grimshaw, BD;
(2015)
Developing a universal T cell for use in adoptive immunotherapy.
Doctoral thesis , UCL (University College London).
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Abstract
Adoptive immunotherapy with genetically-engineered T-cells is showing promise in a clinical setting. However, this approach is limited by the requirement to generate autologous, patient-derived, T-cells, which is both costly and time-consuming. To address this, a methodology which would allow the generation of allogeneic universal T-cells would be highly beneficial. In this project, I have attempted to develop a protocol by which the following T-cell modifications will be introduced: (1) a chimeric antigen receptor (CAR), (2) a sort-suicide gene, (3) human leukocyte antigen (HLA) knock-down, (4) T cell receptor (TCR) knock-down (5) natural killer (NK) cell inhibition. Various strategies have been tested to attempt these modifications. Firstly, I knocked down HLA Class I by using two viral proteins, US11 and infected cell protein (ICP)-47, which give knock-down of 88% and 91% of HLA Class I expression respectively when tested in donor peripheral blood mononuclear cells (PBMCs). In a mixed lymphocyte reaction (MLR) with HLA mis-matched donors, US11 has clearly been shown to reduce the proliferation of effector PBMCs. To address rejection of HLA negative cells by NK cells, HLA-G has been cloned and expressed in cell lines and donor PBMCs to mimic the expression of HLA-G by trophoblast cells in the uterus and by certain cancers. TCR knock-down has been demonstrated in PBMCs, and CD52 knock-down has been achieved in a cell line by using DNA-editing transcription activator-like effector nucleases (TALENs). A CAR can be introduced to redirect the T-cells and a sort-suicide gene can be used to select modified cells and also provide a mechanism to deplete therapeutic cells in case of an adverse event. Some of these strategies have been combined with each other, with limited success.
| Type: | Thesis (Doctoral) |
|---|---|
| Title: | Developing a universal T cell for use in adoptive immunotherapy |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| UCL classification: | UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1470207 |
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