PBMC (500,000 cells per well in 0.5 mL culture medium + PHA) were incubated for 48 h with nanoparticles (25 µg/mL, non cytotoxic concentration) in triplicate wells and, in parallel samples, no NPs were added to set the baseline levels. Cell culture supernatants were harvested and stored at −70 °C until analyzed for the quantifications of different cytokines (Interleukin-17 (IL-17), IL-23, Interferon- γ (IFN-γ), IL-10, IL-13, IL-1-β, IL-2, IL-6, Tumor Necrosis Factor- α (TNF-α) by enzyme-linked immunosorbent assay (ELISA) MAP Human Cytokine reagents and the Milliplex platform (Millipore, Darmstadt, Germany), according to the protocol provided by the manufacturer.

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