Cytometric Bead Array (CBA)
Using the BD Cytometric Bead Array (CBA) Human Kit Flex Set (BD Bioscience-Pharmingen, San Diego, CA). to measure IL-2 (catalog #558270), IL-6 (catalog #558276), IFN-γ (catalog #558269), IL-10 (catalog #558274), TNF (catalog #560112), IL-1β (catalog #558279), IFN-α (catalog #560379), GM-CSF (catalog #558335) and IL-8 (catalog #558277). The kit contains three bead populations with distinct fluorescence intensities. The beads were coated with capture antibodies specific to IL-2, IL-6, IFN-γ, IL-10, TNF, IL-1β, IFN-α, GM-CSF and IL-8 and mixed with phycoerythrin (PE) conjugated detection antibodies. These capture beads were incubated with recombinant standards or test samples to form sandwich complexes. The CBA was analyzed at FACSCcanto flow cytometer (BD Bioscience-Pharmingen). Results were anayzed using graphic and tabular formate using CBA analysis software (BD Bioscience-Pharmingen).