“…For differentiation into DCs, monocytes were incubated in the presence of 100U/mL recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF; Peprotech, Rocky Hill, NJ, USA) and 1,000U/mL recombinant human interleukin-4 (rhIL-4; Peprotech) as described above 17,18 . For phenotypic evaluation, cells were washed with 200µl PBS containing 0.1% bovine serum albumin and 0.01% sodium azide and stained with fl uorescein isothiocyanate (FITC)-conjugated monoclonal antibodies against cluster of differentiation 14 (CD14) and peridinin-chlorophyll protein-cyanine dye (PerCPCy5.5)-conjugated CD209, also known as dendritic cell-specifi c intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) [19][20][21] , on ice for 60min. Cells were washed and analyzed using an Accuri C6 fl ow cytometer (Accuri Cytometers Inc., Ann Arbor, MI, USA) and the FlowJo TM software (Tree Star, Ashland, OR,USA).…”