Cytokine 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).

K15050D V–PLEX Cytokine Panel 1 human kit, K15049D V–PLEX Pro–inflammatory Panel 1 human kit, K15190D V–PLEX Angiogenesis Panel 1 human kit and K151JFC human TIMP–1 kit (Meso Scale Discovery, Rockville, MD, USA) were used to assay interleukin 7 and 8 (IL–7, IL–8), vascular endothelial growth factor C (VEGF–C), tumor necrosis factor alpha (TNF–α) and circulating vascular cell adhesion molecule 1 (sVCAM–1) in plasma samples from patients and HCs according to the manufacturer’s protocol. Meso Scale Discovery Sector Imager 2400 was used to read plates while Meso Scale Discovery Workbench 2.0 software was used to analyze the data.