U plex human il 18 assay

Venous blood samples were collected within 48 h of admission and included basic blood work such as complete blood count, C-reactive protein (CRP), coagulation factors, procalcitonin, kidney function, liver function, diabetes biomarkers, and lipid status.
For analysis of cytokines, blood samples were drawn in EDTA tubes and kept on ice until centrifuged at 3000 g for 15 min at 4 °C. Plasma was separated from the cells and stored at −80 °C until analysis by human pro-inflammatory kits (V-PLEX Viral Panel 3 Human Kit, Meso Scale Discovery). This involved concentration of interleukin (IL)-5, interferon (IFN)-γ, IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor (TNF)-α. The concentration of IL-18 was measured by IL-18 immunoassay (U-PLEX Human IL-18 Assay, Meso Scale Discovery). The concentration of adiponectin was measured by adiponectin immunoassays (Human Adiponectin Kit, Meso Scale Discovery). All cytokines were performed in duplicates with the average presented.

Serum samples were monitored using the V-plex Proinflammatory panel 1 Human kit (Meso Scale Discovery, ref: K15049D) according to the manufacturer’s instructions, for the measurement of IFNγ, IL1β, IL2, IL4, IL6, IL8, IL10, IL12p70, IL13, and TNFα. Soluble Calprotectin (diluted 1:100), IFNα2a and IL18 were analyzed using a R-plex Human Calprotectin Antibody Set (Meso Scale Discovery, ref: F21YB), the ultra-sensitive assay S-plex Human IFNa2a kit (Meso Scale Discovery, ref: K151P3S) and the U-plex Human IL18 assay (Meso Scale Discovery, ref: K151VJK), respectively, following manufacturer’s instructions. Acquisitions and analyses of soluble cytokines and calprotectin were performed on a MESO QuickPlex SQ120 reader and the MSD’s Discovery Workbench 4.0. Each serum sample was assayed twice with the average value taken as the result.