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. 2019 May 20;11(5):696. doi: 10.3390/cancers11050696

Figure 3.

Figure 3

TETARs specifically produced cytokines after antigen encounter. (A,B) CD8+ T cells were lentivirally transduced with a gp100-specific TCR (TCR only) and electroporated with mRNA coding for the CSPG4-specific CAR (TETARs). Non-transduced T cells were either transfected without mRNA (mock) or with CSPG4-specific CAR mRNA (CAR only). Mock-transfected cells were used as a negative control. Following overnight stimulation of T cells with either gp100 peptide-loaded or unpulsed T2.A1 (HLA-A2+, CSPG4−, gp100−) and A375M (HLA-A2+, CSPG4+, gp100−) tumor cells, the production of cytokines was measured in a cytometric bead array (CBA). (A) Specific secretion of tumor necrosis factor (TNF) and (B) interferon-gamma (IFNγ) were calculated by subtracting the number of produced cytokines after co-incubation with unpulsed T2.A1 target cells, which served as a negative control. (A,B) Mean values of four independent experiments with SEM are shown. The p-values were calculated by unpaired Student’s t-test and are listed in Table S2. ** p ≤ 0.01, * p ≤ 0.05.