Meso quickplex sq 120 imager

Colon tissue was homogenized, and cytokine protein levels were measured using a Mouse Pro-inflammatory 7-Plex kit (Meso Scale Diagnostics, Rockville, MD) read on a MESO QuickPlex SQ 120 imager (Meso Scale), as described in detail in the supplemental methods.

Cytokine and chemokines levels were determined by the V-PLEX multiplex panel from Meso Scale Discovery (Rockville, Massachusetts USA). These analytes were measured from serum obtained at the time of registry enrolment, the only time point for which samples are routinely banked for these study participants. Following sample collection, specimens were processed and stored at −70°C until time of measurement. Thirty analytes were examined at enrolment: IL-1α, IL-1β, IL-1 receptor antagonist (RA), IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL-16, IL-17α, IL-23, IL-27, interferon (IFN)-γ, granulocyte-macrophage colony-stimulating factor, macrophage-derived chemokine (MDC), MCP-1, MCP-4, macrophage inflammatory protein (MIP)−1α, MIP-1β, MIP-3α, Vascular Endothelial Growth Factor, eotaxin and TNF-α. Assays were performed as per manufacture protocols and analysed on the MESO QuickPlex SQ 120 imager (Meso Scale Discovery). From banked serum, a second-generation commercial anti-cyclic citrullinated peptide antibody and high sensitivity CRP (hsCRP) were also measured from banked enrolment serum as previously reported.^{26 (link)}