- Author
-
J.J. Freen-van Heeren
- Title
- FISHing out the good guys
- Subtitle
- Enhancing T cell effector function
- Supervisors
-
R.A.W. van Lier
M.C. Wolkers - Award date
- 10 June 2020
- Number of pages
- 205
- ISBN
- 9789463757102
- Document type
- PhD thesis
- Faculty
- Faculty of Medicine (AMC-UvA)
- Abstract
-
CD8+ T cells are critical to protect us from pathogenic microbes and to keep malignantly transformed cells in check. To do so, they produce various effector molecules, including the prototypical T cell cytokine interferon (IFN)-γ. Cytokine production is (partially) regulated through post-transcriptional events, where regulatory elements in mRNA molecules determine mRNA half-life, location and translational rate. Together, these events influence the protein production by T cells. Here, we show that one class of post-transcriptional regulatory elements, AU-rich elements (AREs), regulate IFN-γ protein production in human and murine T cells. Deleting the AREs in the IFNG gene resulted in increased IFN-γ production by human T cells, and rendered murine T cells refractory to inhibitory signals in a tumor model, which significantly improved anti-tumor responses.
Furthermore, we investigated the pathways engaged upon Toll-like receptor (TLR) costimulation. TLR ligands engage different (post)-transcriptional mechanisms to enhance the cytokine production of both human and murine T cells. To investigate the relation between cytokine mRNA and protein, we here described a novel method for the simultaneous measurement thereof.
Lastly, we generated a novel CRISPR/Cas9-tool to genetically modify human T cells. This method does not make use of viral vectors and is electroporation-free. Therefore, it is potentially suitable for augmenting the effector function of T cells used in cellular therapies.
Together, the work in this thesis provides new tools to study and genetically manipulate T cells, and potentially provides novel angles to enhance T cell effector function for therapeutic applications. - Persistent Identifier
- https://hdl.handle.net/11245.1/12216c34-a0df-4a77-b04f-c73b6882add0
- Downloads
-
Thesis (complete)
Front matter
Chapter 1: General introduction
Chapter 2: Measuring T cell responses by flow cytometry-based fluorescence in situ hybridization
Chapter 3: Combined single-cell measurement of cytokine mRNA and protein in immune cells
Chapter 4: Costimulation through TLR2 and TLR7 differently augments cytokine production of CD8⁺ T cells
Chapter 5: Relieving IFN-γ from post-transcriptional regulation elicits superior anti-tumor T cell responses
Chapter 6: Human T cells employ conserved AU-rich elements to fine-tune IFN-γ production
Chapter 7: Developing TAT-Cas9, a novel electroporation-free tool for CRISPR-mediated genome editing in human T cells
Chapter 8: General discussion
English summary; Nederlandse samenvatting; List of co-authors and their contributions; PhD portfolio; List of publications; Curriculum vitae; Acknowledgements
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