Description
- Abstract:
- Abstract of Optimizing T cell/Antigen Presenting Cell coculture conditions for analysis of signaling from the T cell receptor and Major Histocompatibility Complex, by Tobias Robert Hildebrandt, Degree ScM., Brown University, May 2023. T-cell receptor ligation of peptides displayed on the Major Histocompatibility Complex (MHC) of Antigen Presenting Cells (APC) initiate signaling events. Several studies reported canonical signaling events upon engagement of the T-cell receptor complex, yet MHC reverse signaling remains understudied. Here, we describe an optimized approach to permit the study of proximal signaling in live T-cells and APCs simul- taneously via coculture of CD8+ Jurkat T-cells expressing an OT-1 receptor capable of ligating Ovalbumin with a B-cell X T-cell hybrid expressing an empty MHC capable of binding to Ovalbumin in solution. A protein of the MAPK/ERK pathway was used as readout for early signaling. It was shown that only in the presence of Ovalbumin, T-cells with an OT-1 receptor and cells with an empty MHC Complex can fully activate T-cell signaling. Methods such as the one described in this body of work permits the simultaneous study of proximal signaling events in live APCs and T-cells, allowing for the characterization of reverse MHC signaling in different cell types. This method also permits the study of signaling events unique to cell therapies, including signaling in Chimeric Antigen Receptor T-cells (CAR-T), in an in vitro model that closely resembles the human’s body biology.
- Notes:
- Thesis (Sc. M.)--Brown University, 2023
Citation
Hildebrandt, Tobias Robert,
"Optimizing T cell/Antigen Presenting Cell coculture conditions for analysis of signaling from the T cell receptor and Major Histocompatibility Complex"
(2023).
Biotechnology, Biology and Medicine Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:uzepnttc/
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Biology and Medicine Theses and Dissertations
Theses and Dissertations for the Biology and Medicine department....