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Regulatory T-cells are crucial for maintaining immune homeostasis and preventing the development of autoimmune and inflammatory diseases. However, they are also responsible for hindering the immune response against malignancies. Despite years of research, many questions remain unanswered regarding the function of human T-regs. Specifically, what are the biophysical factors that make certain responses susceptible to regulation while others circumvent them. To address those questions, we have established a 3D collagen-based culture that supports the migration of human immune cells. We confer specificity to T-cells using mRNA electroporation to express specific T-cell receptors. Next, using a library of altered peptide ligands with different affinities, we perform time-lapse microscopy and functional analysis in the 3D system. Using this system, we are currently investigating the role of antigen stimulation in T-reg suppressive function. Furthermore, we aim at delineating which inhibitory mechanisms are dominant under different homeostatic and inflammatory conditions where we hypothesise that the dynamics of the T-cell/DC interactions and the strength of the integrated signals will require different suppressive capacity of T-regs. We aim to use our data to construct a mathematical model to describe the mechanism of action of T-regs.



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