Fig. 3 | Nature Communications

Fig. 3

From: Synthetic TRuC receptors engaging the complete T cell receptor for potent anti-tumor response

Fig. 3

Characterization of TRuC-T cell cytotoxicity and cytokine production in vitro. a Tumor cell lysis of luciferase-expressing Nalm6 (Nalm6-LUC) tumor cells after 24 h at a T cell-to-target ratio of 1:1. Data represent results from three independent experiments. b Kinetics of lysis of HeLa cells expressing a truncated form CD19 (HeLa-CD19t) lacking the cytoplasmic domain. T cells and target cells were co-cultured at a ratio of 1:3. HeLa-CD19t lysis was measured in an impedance-based real-time cytotoxicity assay. Representative data of three experiments are shown. c Impact of donor variability on target killing of TRuC and CAR-T cells was assessed as in b. T cells derived from three different blood donors were transduced with TRuC or CAR variants and co-cultured at an effector-to-target ratio of 1:1. d CD107a degranulation by TRuC-T and CAR-T cells upon exposure to CD19-negative or CD19-positive tumor cells. Engineered and non-transduced (NT) T cells were co-cultured with the K562 (light grey), RPMI-8226 (dark gray), Raji (light blue), or Nalm6 (dark blue) at an effector-to-target ratio of 10:1. Error bars are standard deviation. Data represent results from two independent experiments. Samples in ad were measured in triplicates. em Secretion of cytotoxic proteins or cytokines by non-transduced T cells (grey), ε-TRuC (light blue), γ-TRuC (dark blue), 28ζ CAR (red), or BBζ CAR-T cells (purple) following 24-hour co-culture with Nalm6 (1:1 effector-to-target ratio). After incubation, culture supernatants were collected and each analyte was quantified using a Luminex-based immunoassay. Data points represent individual donors (n = 5). Bars indicate group means. Statistical analysis was performed on log-transformed data using parametric one-way ANOVA followed by Dunnett’s (ef) or Tukey’s (gm) multiple comparison test comparing all groups to the non-transduced control group (ef) or all groups against each other (gm). Overall P-value was <0.001 for (em); * Adjusted-P ≤ 0.05, ** Adjusted-P ≤ 0.01, *** Adjusted-P ≤ 0.001, **** Adjusted-P ≤ 0.0001

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