Fig. 4

Antigen-specific cytokine production of receptor-transfected γ/δ T cells and bulk T cells. PBMC were activated with ZA or OKT3 and expanded as mentioned above (Fig. 1). After that, these cells were electroporated with RNA coding for a gp100/HLA-A2-specific TCR or with RNA encoding a MCSP-specific CAR. T cells electroporated without RNA (mock) or with RNA coding for a CEA-specific CAR were used as negative controls. Four hours after electroporation, T cells were co-incubated over night with human tumor cell lines at a 1:1 ratio. Induced cytokine production was examined using intracellular cytokine staining. Double stainings for γ/δ (a) or CD3 (b) and TNF (a + b; left panels) or IFNγ (a + b; right panels) are depicted and percent positives are shown. Presented plots are representatives out of 4 independent experiments. a TNF (left panels) and IFNγ (right panels) production of transfected ZA-expanded γ/δ T cells was characterized in response to co-culture with T2.A1 cells (upper rows) or gp100-pulsed A375M melanoma cells (lower rows). b TNF (left panels) and IFNγ (right panels) production of transfected OKT3-expanded bulk T cells was characterized in response to co-culture with T2.A1 cells (upper row) or gp100-pulsed A375M melanoma cells (lower row)