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Pilot In Vitro Study of Dexamethasone-Induced Immunomodulation in Murine Splenic T Cells
(Tartu Ülikool, 2025) Tyran, Violetta; Chinna Susan Philip; Tartu Ülikool. Loodus- ja täppisteaduste valdkond; Tartu Ülikool. Bioinseneeria instituut
Glucocorticoids (GCs) are key hormones in stress response and chronic elevations from chronic stress or prolonged therapeutic use accelerates immune aging and dysfunction. To dissect their direct, time- and dose-dependent effects on T cells, we established an in vitro model in which murine splenocytes were cultured with graded concentrations of dexamethasone (a potent, synthetic GC) and analyzed by 22-color spectral flow cytometry over 24, 48, and 72 hours. Dexamethasone caused a progressive, dose-dependent decline in overall viability and absolute CD4⁺/CD8⁺ T cell counts, transiently increased proliferation (Ki-67) at 48 hours before inducing collapse and apoptosis, and modulated activation (CD69, CD25) and exhaustion (PD-1) markers. Memory and naïve T cell subsets were especially vulnerable, whereas low-dose dexamethasone selectively enriched regulatory (CD4⁺Foxp3⁺) and Th1 (CD4⁺Tbet⁺) subsets. Effector cytokine production (IFN-γ, TNF-α) was markedly suppressed in a dose- and time-dependent manner. This platform thus provides a robust tool for exploring GC-driven immunosenescence, GC resistance mechanisms, and strategies to restore T cell function after chronic GC exposure.