Chemokine panel 1 human kit

From each blood sample for each time point 250 µl of the cell suspension was centrifuged at 4 °C for 5 min with 1,000 G. The supernatant was collected and frozen for cytokine and chemokine determination at − 80 °C. Levels of IL-1β, IL-2, IL-4, IL-6, IL-8, IL10, IL-12p70, IL-13, TNF-α, and IFN-γ (V-PLEX Proinflammatory Panel 1 Human Kit), Eotaxin, Eotaxin-3, IL-8, IL-8 (HA), IP-10, MCP-1, MCP-4, MDC, MIP-1α, MIP-1β, and TARC (V-PLEX Chemokine Panel 1 Human Kit) and GM-CSF, IL-1α, IL-5, IL-7, IL-12/IL-23p40, IL-15, IL-16, IL-17A, TNF-β, and VEGF-A (V-PLEX Cytokine Panel 1 Human Kit) were analysed via an electrochemiluminescent immunoassay according to the manufacturer’s instructions (Meso Scale Discovery (MSD), Gaithersburg, MD, USA) using the Meso Quick Plex SQ 120. Raw data were analysed using the Discovery Workbench 4.0 software (MSD).

Medium was collected from the cell cultures at the time points indicated and stored at −80 °C. Samples were thawed and centrifuged to remove any debris on the day of analysis. Chemokines were quantified in the supernatants using the V-PLEX Chemokine Panel 1 Human Kit, on the QuickPlex Meso SQ 120 (Meso Scale Diagnostics, LLC), according to the kit manuals.