Figure 3.

Chimeric antigen receptor (CAR) triggered cytokine secretion pattern of CAR T cells is specifically altered by MSCs. (A) Anti-carcinoembryonic antigen (CEA) CAR T cells were co-cultivated in 96-well plates (2.5 × 10⁴ cells/well) in quadruples with LS174T tumor cells (2.5 × 10⁴ cells/well), respectively. After 48 h supernatants were recovered and supernatants of 2 adjacent wells were pooled and subjected to 43-parameter multiplex analysis as described in Materials and Methods. Mean values of duplicates were determined and data were displayed as heat map. (B–D) Anti-CEA CAR T cells were co-cultivated with LS174T cancer cells as described above in presence of wt or IL7/IL12 modified MSCs (5 × 10³ cells/well) and subjected to 43-parameter multiplex analysis as described above. The number of secreted cytokines and growth factors was determined, and x-fold secretion compared to release of cytokines in presence of CAR T cells and tumor cells alone was calculated and presented as heat map (B). Cytokine secretion was dependent on the presence of MSC (C) or IL7/12 producing MSCs (D), data indicate mean values of duplicates.