Luminex xponent 4

A Luminex IS100 machine was used to detect cytokines with the MILLIPLEX MAP mouse cytokine/chemokine, premixed, 10 Plex kit. Murine T cells were cocultured with tumor targets at a 1:1 ratio for 24 hours in a 96-well round-bottom plate in 200 μl of media. The supernatant fluid was collected and analyzed for cytokines on a Luminex IS100 instrument. Luminex FlexMap3D system and Luminex Xponent 4.2 (Millipore Corporation) were used to detect the cytokines.

To analyze in vitro and ex vivo T-cell cytokine production, T cells and antigen positive tumor cells were cocultured in a 1:1 ratio for 24 hours in a 96-well round-bottom plate in 200 μl of media. The supernatant was collected and analyzed for cytokines on a Luminex IS100 instrument. Luminex FlexMap3D system and Luminex Xponent 4.2 (Millipore Corporation) were used to detect cytokines.

Cytokines were detected in the supernatant using the Luminex FlexMap3D system (Millipore Corporation, Billerica, MA, USA). Cytokine concentrations were assessed using Luminex Xponent 4.2 (Millipore Corporation).
T-cell cytotoxicity was assessed by 4 h chromium release assays, as previously described.^{24 (link)}To assess expansion, CAR T cells were co-cultured with Set2 or Nalm6 cells in the absence of supplemented cytokines. The cell count for each sample was assessed at indicated times by flow cytometry and cells were enumerated using 1,2,3 count ebeads (ebiosciences, San Diego, CA, USA) according to the manufacturer’s instructions.