TIMP-3–treated DCs altered the Th1/Th2 polarization. To stimulate the naive T cells, 4 types of mDCs were applied: (1) imDCs stimulated with LPS (Control); (2) imDCs stimulated with LPS plus TIMP-3; (3) imDCs preincubated with 500nM wortmamin for 1 hour with the following maturation by LPS plus TIMP-3 (wortmamin + TIMP-3); and (4) TIMP-3–knockdown imDCs stimulated by LPS (si-TIMP-3). These mDCs were mixed with allogeneic naive T cells for 6 days to induce T-cell polarization at a DC:T ratio of 1:5. (A) Dot plots of IFN-γ and IL-4 production by CD4⁺ T cells from one representative donor. For intracellular analysis of cytokine production, CD4⁺ T cells were restimulated with 10 ng/mL phorbol myristate acetate and 1 μg/mL ionomycin in the presence of 10 μg/mL brefeldin A for 5 to 6 hours. Cells were then fixed, permeabilized, and stained for IFN-γ and IL-4 and analyzed by flow cytometry. (B) Average of percentage of CD4⁺ T cells positive for IFN-γ and IL-4 staining from 4 individual donors are presented in the bar charts. (C) Supernatants of the mixed lymphocyte reaction coculture were harvested, and the production of IFN-γ, IL-4, IL-5, IL-10, and IL-13 was detected by the Bio-Plex Protein Array system. Data are the mean ± SD from 3 independent experiments. *P < .05.