Tgfbmag 64k 03

After an overnight fasting for at least 10 hours when DK/DKA resolved, fasting blood samples were collected and centrifuged, and sera were collected for cytokine determination. The following cytokines were measured: interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A/CTLA-8, IL-17F, IL-21, IL-22, IL-23, interferon-γ (IFN-γ), programmed death-1 (PD-1), PD ligand 1 (PD-L1), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4/CD152), CD40, transforming growth factor β1 (TGFβ1), TGFβ2, and TGFβ3. All these measurements of cytokines were performed using Milliplex MAP technology in the Luminex platform (Millipore, Billerica, MA). Immunology multiplex assay kits (HTH17MAG-14K-12, TGFBMAG-64K-03, and HCKPMAG-11K-04) were obtained from MERCK (MERCK Millipore Co., Billerica, MA, USA). All the collected serum samples were mixed and diluted at a ratio of 1:2 with buffer, and all samples, standards, and quality controls were assayed in accordance with the manufacturer’s instructions. Median fluorescent intensity (MFI) data were acquired using MILLIPLEX Analyst.V5.1 software, and a 5-parameter logistic method was used to calculate cytokine concentrations.