Protocartaplex analyst 1

According to the manufacturer’s recommendations, we used the mouse ProtocartaPlex 10-plex immunoassay kit (Thermofisher Scientific, Vienna, Austria) to measure cytokine levels in serum samples. We classified the immune mediators into five categories: proinflammatory mediator (Monocyte Chemoattractant Protein [MCP]–1), type 1 cytokines (IL-2, interferon [IFN]- γ, and tumor necrosis factor [TNF]–α), type 2 cytokines (IL-4, IL-5, IL-10, and IL-13), a T helper 17 cytokine (IL-17), and a regulatory cytokine (Transforming Growth Factor [TGF]-β1). Data were acquired on the MAGPIX-Luminex XMAP technology (Merck Millipore, Darmstadt, Germany) using the Luminex XPONENT software solutions. The concentration (pg/mL) of each cytokine was obtained by interpolating MFI (median fluorescent intensity) of a dilution standard curve over seven dilution points supplied with the kit and calculated by the ProtocartaPlex Analyst 1.0 software (eBioscience, Thermo Fisher Scientific, USA).

We measured cytokine levels in serum samples using the mouse ProtocartaPlex 10-plex immunoassay kit (Thermofisher Scientific, Vienna, Austria), following the manufacturer’s recommendations. Data were acquired on the Luminex XMAP technology (Merck Millipore, Darmstadt, Germany) using the Luminex XPONENT software solutions. The concentration (pg/mL) of each cytokine was determined by interpolating the median fluorescent intensity (MFI) of a dilution standard curve over seven dilution points supplied with the kit and calculated by the ProtocartaPlex Analyst 1.0 software (eBioscience, Thermo Fisher Scientific, USA). The immune mediators were classified into four categories: T helper-1 cytokines (IL-2, IFN-γ, and TNF-α), T helper-2 cytokines (IL-4, IL-5, and IL-13), T helper-17 cytokine (IL-17A), and regulatory cytokines (TGF-β1 and IL-10).