Legendplex human th cytokine panel 13 plex kit

Following 42–44 h of stimulation, the supernatant of the PBMC cultures was collected and frozen at −80°C for later use. The supernatant of six D2R‐specific T‐cell‐positive patients was analysed for secreted cytokines with the LEGENDPlex Human Th Cytokine Panel 13‐plex Kit (BioLegend, San Diego, CA, USA) and an IFN‐γ enzyme‐linked immunosorbent assay (ELISA; Mabtech Stockholm, Sweden) used as per the manufacturer's protocol. The following cytokines were quantified: IL‐2, IL‐4, IL‐5, IL‐6, IL‐9, IL‐10, IL‐13, IL‐17A, IL‐17F, IL‐21, IL‐22, IFN‐γ and TNF. All samples were measured undiluted and acquired on BD LSR II and BD FACSCanto II. Data were analysed with LEGENDPlex Data Analysis Software v8 (BioLegend), Excel and GraphPad Prism.

Serum human cytokine concentrations were determined using a LEGENDplex Human Th Cytokine Panel (13-plex) kit (BioLegend) according to the manufacturer’s instructions with an Attune NxT Flow Cytometer and Autosampler (Thermo Fisher Scientific). Cytokine concentrations outside the standard curves were set to the corresponding detection limits for statistical analyses.