Authors:

Oluwagbemisola Awonusonu

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About Paper:

In healthy individuals, a subset of regulatory T cells (Tregs) suppresses self-reactive immune cells, thereby protecting pancreatic beta cells. Our work seeks to enhance the functionality of Tregs in the context of T1D by using gene-editing to enhance Treg activation and migration to sites of inflammation.To accomplish this, lentiviral transduction was used to generate Tregs expressing a preproinsulin (PPI)-directed T cell receptor (TCR). Following 14 days of in vitro expansion, PPI-TCR+ Tregs were characterized using flow cytometry to assess activation and lineage markers. After 24 hours, we observed increased activation in Tregs expressing moderate and high-affinity PPI-TCRs, whereas low-affinity PPI- TCR+ Tregs showed no differences from PPI-TCR (polyclonal) Tregs. We further examined the proliferative capacities of these PPI-TCR+ Tregs after 4 days of antigen-specific stimulation and identified significant increases in proliferation for PPI-TCR+ Tregs compared to polyclonal Tregs. Finally, we used in vitro suppression assays to measure CD8+ and CD4+ T responder proliferation by flow cytometry and observed that PPI-TCR Tregs demonstrated improved suppressive capacity compared to polyclonal Tregs after four days of co-culture.This data suggests PPI-specific TCRs can improve in vitro Treg suppressive and proliferative capacity during antigen-specific stimulation with potential implications for adoptive cell immunotherapies.

Source:

University of Florida / Oluwagbemisola Awonusonu, Alexander Pearce, / 2023

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Co-authors:

Oluwagbemisola Awonusonu