Polyinosinic acid poly 1

Following 6 days in culture, immature MoDCs were plated at 5x10 4 cells/well in 96-well roundbottom plates (Corning). For the basic uptake assay, PLGA nanoparticles loaded with OVA-Alexa Fluor 488 and matched by polymer weight were added to cells, which were incubated for 60 min at 37 o C or 4 o C before washing 3 times (930 x g for 2 minutes) with PBS. For the inhibition studies, cells were pre-treated with; mannan (MAN; 50 µg/ml), polyinosinic acid (poly I; 50 µg/ml) or amantidine (50 µg/ml) (all Sigma) for 30 min at 37 o C. Uptake was assessed using a MACSQuant Analyzer flow cytometer. Dead cells were excluded by staining with Near-Infra Red Fixable Live/Dead Cell stain (Life Technologies), and the mean Alexa Fluor 488 fluorescence intensity of viable cells used as a measure of uptake.