Flow Cytometry Analysis

Cells were incubated with FcR blocking reagent for 5 min followed by incubating with or without the mixture of fluorescent-labeled antihuman monoclonal antibodies: CD14-ECD, CD15-PE-Cy5, CD3-fluorescein isothiocyanate (FITC), and IFN-γ-PE (Beckman Coulter, Brea, CA, United States) for 30 min. For intracellular staining, cells were treated with IntraPrep Permeabilization Reagent (Beckman Coulter, Brea, CA, United States) before the staining. The Annexin V-FITC and propidium iodide were used to eliminate the dead cells from the analysis. Auto-fluorescence and non-specific staining were determined by using isotype-matched controls. The analysis was performed using a Cytomics FC 500 flow cytometer (Beckman Coulter, Brea, CA, United States).