Fig. 2.

Binding of MZF by DCs requires β-glucan and mannan recognition. a Internalization of Fl-MZF by DCs. DCs were cultured in RPMI-1640 medium containing 10% FBS with Fl-MZF (400 μg/ml) at 37°C. Cells were collected at various time periods and then washed, fixed, and observed by fluorescence microscopy. b Binding of MZF by DCs. DCs (2 × 10⁵ cells/tube) were incubated on ice for 1 h in PBS containing 3% FBS and Fl-MZF at various concentrations. After incubation, the cells were washed and fixed. Fl-MZF⁺ DCs were identified by flow cytometry. c Laminarin, mannan, and anti-Dectin-1 Ab inhibited the DC recognition of Fl-MZF. Ice-cold PBS containing 3% FBS with or without inhibitors (laminarin, mannan and dextran; 1 mg/ml), anti-Dectin-1 Ab (2A11), or isotype Ab (rat IgG2bκ) was added to the DCs (2 × 10⁵ cells/tube) and incubated on ice for 1 h before the addition of Fl-MZF (400 μg/ml). Cells were incubated on ice for 1 h, washed, and fixed. Fl-MZF⁺ DCs were identified by flow cytometry. The data shown have been normalized to the percentages of Fl-MZF⁺ CD11c⁺ cells of uninhibited control. Values are expressed as mean ± SE, n = 3; *p < 0.05