Cytokine multiplex assay
REGRESSION
Quantification of cytokines (for example, IL-6, IFNɣ) in serum was performed using the Meso Scale Discovery (MSD) 10-plex assay using the Proinflammatory Panel 1 Multiplex Test Kit. IFNɣ inducible protein 10 was measured using the Chemokine Panel 1 Kit (MSD). Assays were performed per the manufacturer’s protocol with a seven-point standard curve generated using a fourfold dilution series in triplicate. The standards were used as described in the package insert. All samples were evaluated in duplicate at standard 1:2 dilution and maximal 1:8 dilution; calculated percentage coefficients of variation for the duplicate measures were less than 20%. Data were acquired on a Quickplex SQ120 Imager instrument (MSD) and analyzed using an MSD Discovery Workbench and a four-parameter logistic model. Reported values included those within the standard curve range and those calculated by the logistic regression analysis.