Figure 6: Apoptotic T cell-induced tolerogenic DCs block development of CD8+ regulatory T cells. CD11c+ DCs were isolated from spleen of C57 BL/6J mice and sorted using flow cytometry. CD11c+ DCs were incubated overnight with apoptotic or fresh T cells at 37°C. MOGspecific CD4+CD25- T cells were isolated from 2D2 mice and co-cultured with DCs pulsed with 0.1 μM MOG peptide and treated with apoptotic or fresh T cells as a control for 3 days at 37°C. A flow cytometry assay was done. CD4-CD8+ cells were gated. Expression of FoxP3 (A), IL-10 (B), CD25 (C) and CD122 (D) is shown. Isotype controls are also demonstrated. Error bars represent mean and SD of triplicate determinations of percentage (%) of expression of Treg-associated markers on CD4-CD8+ T cells in three independent experiments (n=3, t test, *P< 0.05).