Fig. 4

BsAb-armed T cells could be easily adjusted the cell surface BsAb level and reduce cytokine release significantly. A The T cells were armed with different concentrates (0.32–200 nM) of anti-PSMA/anti-CD3 BsAbs. The surface BsAb level were detected by staining anti-His antibody and (B) measured as MFI by flow cytometry. C For testing the self-releasing cytokines of T cells, overactivated T cell (OKT3 co-cultured T cell), and BsAb-armed T cells, serial dilution of OKT3 or BsAbs were added to T cells for 1 h. After washed, the T cells, overactivated T cell or BsAb-armed T cells were incubated for 16 h at 37 °C. The supernatants were harvested for testing by commercial ELISA kits for human IL-2, TNF-α and IFN-γ. D The T cells arming with 0.4, 4, or 40 nM of each BsAb or OKT3 cultured T cell were applied at different ratios (effector cell/target cell ratios of 3:1, 5:1, or 10:1) and added to LNCaP. The anti-cancer efficiency was analyzed by cytotox 96 non-radioactive cytotoxicity assay kit. E The BsAb-armed T cells or BsAb mixed T cells were incubated with LNCaP for 16 h. The anti-cancer efficiency was analyzed by cytotox 96 non-radioactive cytotoxicity assay kit. Bar, SD; ***p < 0.001