Figure 6

Effect of treatment with different microtubule-depolymerizing agents on expression of cell-surface markers and CD4⁺ and CD8⁺ T-cell proliferation in mouse bone marrow-derived immature dendritic cells. (A) Expression of surface markers on DCs. Cells were treated for 24 h with CMQ (0.1, 0.5 and 1 μM), colchicine (2.5 μM) and LPS (1 μg/ml), and harvested. Examination for the expression of CD40, CD80, CD86, MHC class II and CD11c markers was performed by flow-cytometry. The levels of CD40, CD80, CD86, MHC class II were expressed as mean fluorescence intensity (MFI). (B) CD4⁺ and CD8⁺ T-cell proliferation. After indicated treatments, treated DCs were co-cultured with CD4⁺ T cells or CD8⁺ T cells in a ratio of 1:20 (DCs versus CD4⁺ T cells or CD8⁺ T cells) for 72 h. T-cell proliferation was determined in vitro using a BrdU proliferation ELISA kit (Roche, Heidelberg, Germany) according to the manufacturer's instructions. The T-cell proliferation was expressed as the stimulation index, the OD₄₅₀ value of co-culture of treated-DCs and T cells was divided by the value of co-culture of DMSO-treated-DCs co-cultured with T cells. All Data were expressed as mean ± S.D. P values less than 0.05 were considered statistically significant (*, P < 0.05; **, P < 0.01; ***, P < 0.001).