Fig. 3

DARPin28z induces human CAR-T cell cytokine production upon HER2 stimulation. 10⁶ scFv28z, DARPin28z, or CAR –’ve transduced human T cells were stimulated with HER2 (HER2Fc fusion protein) or an unrelated target (KDRFc fusion protein) for four hours at 37 °C in a 96-well plate. a Production of IFN-γ and TNF-α was measured by ICS and subsequent flow cytometry. Data from CD8⁺ NGFR⁺ T cells is presented as mean ± SD. Bracketed numbers are quantitative of the representative plots shown. b Production of IL-2 by CD8⁺ NGFR⁺ T cells as measured by ICS. c Production of CD107a by CD8⁺ NGFR⁺ T cells as measured by ICS. d Pie graphs capturing the distribution of single and multi-functional CAR-T cells as produced with SPICE software. Pie arcs indicate functional populations represented by pie wedges. Error bars = SD. * = p < 0.05 ** = p < 0.005 *** = p < 0.001. All data from n = 3 experiments repeated with T cells from two donors