Figure 6.
Figure 6. In vitro impacts of microtubule depolymerizing drugs on DC maturation. (A) BM-DCs were incubated for 24 hours with COL (3 μg/mL), PDP (1 μg/mL), or vehicle alone and then examined for the surface expression of MHC II, CD40, and CD80 and for cell viability by PI uptake (mean ± SD; n = 3). (B) The supernatants of the same BM-DC cultures were examined for the secretion of the indicated cytokines (mean ± SD; n = 3). (C) XS106 DCs were incubated for 5 hours with COL (3 μg/mL), PDP (1 μg/mL), or vehicle alone and then examined for NF-κB activation (mean ± SD; n = 3). (D) BM-DCs generated from BALB/c mice were incubated for 24 hours with 3 μg/mL COL (○) or vehicle alone (•), washed extensively, and then cocultured at the indicated numbers with allogeneic T cells (5 × 104 cells per well) purified from C57BL/6 mice. Data shown are 3H-thymidine uptake on day 4 (mean ± SD; n = 3). (E) BM-DCs were incubated for 24 hours with 3 μg/mL COL (open symbols) or vehicle alone (closed symbols) in the presence (circles) or absence (triangles) of 2 μg/mL OVA323-339 peptide during the last 2 hours of the incubation period. After extensive washing, DCs were cocultured at the indicated numbers with CD4 T cells (5 × 104 cells per well) purified from the DO11.10 transgenic mice. Data shown are 3H-thymidine uptake on day 3 (mean ± SD; n = 3). (F) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were examined for their capacity to uptake FITC-dextran at 4°C (open bars) or 37°C (closed bars). Samples were then examined for FITC fluorescent signals (mean ± SD; n = 3) within the CD11c+ populations. (G) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were stained with anti–CCR-7 mAb (closed bars) or isotype-matched control IgG (open bars). Data shown are fluorescent signals (mean ± SD; n = 3) within the CD11c+ populations. (H) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were examined for their migratory capacity toward CCL-19 (closed bars) or PBS alone (open bars). Data shown are the percentage DC migration (mean ± SD; n = 3). In all panels, statistically significant changes as compared with controls treated with vehicle alone are indicated with asterisks (*P < .05, **P < .01).

In vitro impacts of microtubule depolymerizing drugs on DC maturation. (A) BM-DCs were incubated for 24 hours with COL (3 μg/mL), PDP (1 μg/mL), or vehicle alone and then examined for the surface expression of MHC II, CD40, and CD80 and for cell viability by PI uptake (mean ± SD; n = 3). (B) The supernatants of the same BM-DC cultures were examined for the secretion of the indicated cytokines (mean ± SD; n = 3). (C) XS106 DCs were incubated for 5 hours with COL (3 μg/mL), PDP (1 μg/mL), or vehicle alone and then examined for NF-κB activation (mean ± SD; n = 3). (D) BM-DCs generated from BALB/c mice were incubated for 24 hours with 3 μg/mL COL (○) or vehicle alone (•), washed extensively, and then cocultured at the indicated numbers with allogeneic T cells (5 × 104 cells per well) purified from C57BL/6 mice. Data shown are 3H-thymidine uptake on day 4 (mean ± SD; n = 3). (E) BM-DCs were incubated for 24 hours with 3 μg/mL COL (open symbols) or vehicle alone (closed symbols) in the presence (circles) or absence (triangles) of 2 μg/mL OVA323-339 peptide during the last 2 hours of the incubation period. After extensive washing, DCs were cocultured at the indicated numbers with CD4 T cells (5 × 104 cells per well) purified from the DO11.10 transgenic mice. Data shown are 3H-thymidine uptake on day 3 (mean ± SD; n = 3). (F) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were examined for their capacity to uptake FITC-dextran at 4°C (open bars) or 37°C (closed bars). Samples were then examined for FITC fluorescent signals (mean ± SD; n = 3) within the CD11c+ populations. (G) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were stained with anti–CCR-7 mAb (closed bars) or isotype-matched control IgG (open bars). Data shown are fluorescent signals (mean ± SD; n = 3) within the CD11c+ populations. (H) Following 24 hours of incubation with 3 μg/mL COL or vehicle alone, BM-DCs were examined for their migratory capacity toward CCL-19 (closed bars) or PBS alone (open bars). Data shown are the percentage DC migration (mean ± SD; n = 3). In all panels, statistically significant changes as compared with controls treated with vehicle alone are indicated with asterisks (*P < .05, **P < .01).

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