Regulatory T cells (Tregs) are key to keeping your immune system balanced and preventing autoimmune diseases. Their pronounced immunosuppressive function makes them promising targets for the development of therapeutic approaches to autoimmune diseases and immune rejection of transplants, but their low frequency and the challenges of isolating and proliferating these cells hinder their development. One approach to selectively promote the growth of Treg cells is the use of the antibiotic rapamycin, which partially limits the proliferation of effector T cells by inhibiting the mTOR target and promotes the differentiation of Treg cells, as evidenced by an increased proportion of Tregs in samples. In our thesis, we isolated Treg cells from mouse spleens by negative selection of CD4+ cells and positive selection of CD25+ cells. We cultured the cells for 15 days at different concentrations of rapamycin in the presence of the cytokine mIL-2 and monitored the number and viability of the cells. Finally, we analysed the samples using flow cytometry. We successfully quantified the expression of characteristic surface and intracellular markers and thus determined the size of the Treg population. We observed an increased proportion of Tregs compared to T helper cells (Th) at elevated concentrations of rapamycin (100 nM). Cell viability and number were higher in samples without rapamycin than in samples with rapamycin. The use of rapamycin in human Tregs is already standard practice, but it has not yet been standardised in mouse Tregs. The literature reports both improved and reduced expansion of mouse Treg cells in the presence of rapamycin. We obtained preliminary results with our thesis work, on the basis of which we can plan further experiments to draw more reliable conclusions.
|
