Fig. 5: CARTIV promotors show an additive response only to TNFα and hypoxia in primary human T cells and NK92 cells due to autocrine IFNγ secretion.
From: Synthetic promoters to induce immune-effectors into the tumor microenvironment
a Representative plots of primary human T cells or NK92 cells infected using lenti viral vectors with RFP670 under the control of the indicated CARTIV promotor and ZsGreen controlled by the ef1α core promotor. At least 72 h following infection, the cells were incubated for 48 h with the indicated cytokines at 250 U/mL and for 18 h under hypoxic or normoxic conditions, harvested and analyzed by flow cytometry. Data shown are ZsGreen-positive, single-discriminated, and DAPI-negative results. b, c Geometric mean of RFP670 in ZsGreen-positive cells normalized to unstimulated cells, showing the average of duplicates, an ordinary one-way ANOVA was performed to test for significance, error bars indicate standard deviation of NK92 or human primary T-cells. d Supernatants of primary human T cells incubated for 48 h with 250 U/mL indicated cytokines and for 18 h under hypoxic or normoxic condition were collected, and ELISA for IFNγ was performed. Showing average of triplicates, an ordinary one-way ANOVA was performed to test for significance, error bars indicate standard deviation. Results are from one representative experiment of four performed.
