Glucocorticoid hormone differentially modulates the in vitro expansion and cytokine profile of thymic and splenic Treg cells.

摘要: Abstract Objective Functional disturbances in regulatory T cells (Treg) have been described autoimmune diseases, and their potential therapeutic use is intensively studied. Our goal was to investigate the influence of glucocorticoid hormone on vitro differentiation Treg from thymic splenic CD4+ under different conditions establish methods for generating stable functionally suppressive iTregs future adoptive transfer experiments. Methods Thymic lymphocytes were isolated 3 4 week-old control vivo dexamethasone (DX) pretreated BALB/c mice using magnetic bead negative selection, followed by CD25 positive selection. The cultured with anti-CD3/CD28 beads IL-2 presence or absence TGFβ and/or DX 3–6 days. Multiparametric flow cytometry performed CD4, CD25, CD8, (LAP) cell surface Foxp3, IL-4, IL-10, IL-17 IFNγ intracellular staining. Quantitative RT-PCR measure cytokine Foxp3 mRNA levels. Results Differentiation thymus-derived into iTreg most effective (24–25%) when beads, IL-2, present. Splenic expansion same resulted a higher (44–45%) ratio that further increased (up 50% Treg) DX. Elevated immunosuppressive (IL-10 TGFβ) production could be measured both at protein levels without elevation Th1/Th2 Th17 production. We got highest (74%) CD4+CD25+ stimulated TGFβ. In days pretreatment enhanced expression decreased spleen-derived cells. these Tregs relative IL-10 significantly all stimulation conditions, while level did not change. Conclusion promotes cells, enhances Foxp3+ cytokines vitro. inhibited differentiated hypothesize patients receiving GC therapy may need special attention prior transplantation