Research Article: In vitro generation of ROR?t + regulatory T cells reveals enhanced immunosuppressive function and OXPHOS-dependent metabolism
Abstract:
ROR?t + regulatory T cells (Treg) play a crucial role in immune regulation, particularly in the gut. However, most current knowledge about this subset derives from in vivo studies, as in vitro investigation has been limited by the lack of protocols capable of preserving their phenotype.
Here, we developed and optimized an in vitro differentiation protocol to efficiently generate ROR?t + Treg cells. The protocol was evaluated based on the frequency of ROR?t + Treg cells generated, their suppressive function compared to conventional induced Treg (iTreg), and their metabolic profile.
The optimized protocol increased the frequency of ROR?t + Treg cells in vitro by up to 70%, providing a robust system for their study. Functionally, in vitro –differentiated ROR?t + Treg cells displayed enhanced immunosuppressive activity compared to conventional iTreg, effectively inhibiting effector CD4? T cell proliferation. Metabolic analyses further revealed a reliance on oxidative phosphorylation (OXPHOS) in this subset.
This protocol enables the efficient in vitro generation of ROR?t + Treg cells, facilitating functional and metabolic studies of this population and opening new avenues for potential therapeutic applications in immune-mediated diseases.
Introduction:
ROR?t + regulatory T cells (Treg) play a crucial role in immune regulation, particularly in the gut. However, most current knowledge about this subset derives from in vivo studies, as in vitro investigation has been limited by the lack of protocols capable of preserving their phenotype.
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