Figure 1
Figure 1. The immunostimulatory phenotype and functions of iDCZol+ and mDCZol+ are preserved. (A) Dose-response and time course analyses of Zol activity on iDC viability and cell counts. Total counts of viable iDC per well were determined by annexin-V and propidium iodide staining. Bars represent the mean ± SEM of 3 experiments. Results indicate that iDC exposure up to 10 μM Zol for 72 hours does not exert any direct cytotoxic activity. (B) Zol treatment does not affect (1) the capability of iDC to internalize FITC-dextran (iDCZol− 49 ± 17% versus iDCZol+ 53 ± 11%; n = 3, P > .05); and (2) the downregulation of FITC-dextran uptake of mDC (mDCZol− 17 ± 6% versus mDCZol+ 15 ± 3%, n = 3, P > .05). iDCZol−, iDCZol+, mDCZol−, and mDCZol+ were incubated with FITC-dextran for 2 hours at 4°C (dotted light line histograms) or 37°C (solid bold line histograms). mDCZol− and mDCZol+ were matured with TNF-α and IL-1β. Results are 1 of 3 (mDC) to 6 (iDC) experiments. (C) Zol treatment does not affect the allostimulatory activity of iDCZol+ and mDC Zol+. iDC and mDC subsets were incubated for 5 days with allogeneic PBL cells (allo-PBL). Proliferation was determined on day 5 by 3HTdR incorporation. Bars represent the mean (± SEM) of 3 head-to-head experiments. The proliferation rates of allo-PBL stimulated with iDCZol+ and mDCZol+ were slightly higher than those induced by iDCZol− and mDCZol−, but differences are not statistically signficant (iDCZol− 48 600 ± 25 700 cpm/well versus iDCZol+ 52 900 ± 25 800; mDCZol− 65 300 ± 35 800 cpm/well versus mDCZol+ 73 500 ± 42 800; P > .05). (D) Release of IL-4, IL-6, IL-10, and IL-12 in the supernatants of iDCZol−, iDCZol+, mDCZol−, and mDCZol+. Cytokines were measured with a multiparametric cytometric bead immunoassay and results are expressed as pg/mL. Bars represent the mean (± SEM) of 4 experiments. Closed bars represent iDCZol− and mDCZol−, open bars represent iDCZol+ and mDCZol+. mDCZol− and mDCZol+ were matured with TNF-α and IL-1β. IL-10 production was decreased in iDCZol+, whereas IL-12 was increased and IL-6 was decreased in mDCZol+. None of these differences reached a statistical significance.

The immunostimulatory phenotype and functions of iDCZol+ and mDCZol+ are preserved. (A) Dose-response and time course analyses of Zol activity on iDC viability and cell counts. Total counts of viable iDC per well were determined by annexin-V and propidium iodide staining. Bars represent the mean ± SEM of 3 experiments. Results indicate that iDC exposure up to 10 μM Zol for 72 hours does not exert any direct cytotoxic activity. (B) Zol treatment does not affect (1) the capability of iDC to internalize FITC-dextran (iDCZol− 49 ± 17% versus iDCZol+ 53 ± 11%; n = 3, P > .05); and (2) the downregulation of FITC-dextran uptake of mDC (mDCZol− 17 ± 6% versus mDCZol+ 15 ± 3%, n = 3, P > .05). iDCZol−, iDCZol+, mDCZol−, and mDCZol+ were incubated with FITC-dextran for 2 hours at 4°C (dotted light line histograms) or 37°C (solid bold line histograms). mDCZol− and mDCZol+ were matured with TNF-α and IL-1β. Results are 1 of 3 (mDC) to 6 (iDC) experiments. (C) Zol treatment does not affect the allostimulatory activity of iDCZol+ and mDC Zol+. iDC and mDC subsets were incubated for 5 days with allogeneic PBL cells (allo-PBL). Proliferation was determined on day 5 by 3HTdR incorporation. Bars represent the mean (± SEM) of 3 head-to-head experiments. The proliferation rates of allo-PBL stimulated with iDCZol+ and mDCZol+ were slightly higher than those induced by iDCZol− and mDCZol−, but differences are not statistically signficant (iDCZol− 48 600 ± 25 700 cpm/well versus iDCZol+ 52 900 ± 25 800; mDCZol− 65 300 ± 35 800 cpm/well versus mDCZol+ 73 500 ± 42 800; P > .05). (D) Release of IL-4, IL-6, IL-10, and IL-12 in the supernatants of iDCZol−, iDCZol+, mDCZol−, and mDCZol+. Cytokines were measured with a multiparametric cytometric bead immunoassay and results are expressed as pg/mL. Bars represent the mean (± SEM) of 4 experiments. Closed bars represent iDCZol− and mDCZol−, open bars represent iDCZol+ and mDCZol+. mDCZol− and mDCZol+ were matured with TNF-α and IL-1β. IL-10 production was decreased in iDCZol+, whereas IL-12 was increased and IL-6 was decreased in mDCZol+. None of these differences reached a statistical significance.

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