Fig. 4 | Nature Communications

Fig. 4

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

Fig. 4

TRuC-T and CAR-T cell activation and signaling upon binding to CD19-positive target cells. a T cells were co-cultured overnight in the presence of Nalm6 tumor cells. T cell activation markers CD69 and CD25 were analyzed by flow cytometry. The graph depicts the percentage of CD69/CD25-double positive cells (mean ± SD of triplicates). Statistical analysis was performed on log-transformed data using parametric one-way ANOVA followed by Dunnett’s multiple comparison test comparing means of all groups to the vector control group. Representative data of three independent experiments are shown. b Phosphorylation of CD3ε and LAT in Non-transduced T cells (black), ε-TRuC (light blue), γ-TRuC (dark blue), 28ζ CAR (red), or BBζ CAR (purple) T cells upon co-culture with CD19-positive Raji cells at a 10:1 effector-to-target ratio for 30 min. Protein phosphorylation in cell lysates was analyzed using a Luminex-based cell signaling assay. Data points represent values for individual donors (n = 5) and bars indicate group means. Representative data of two experiments are shown. P-values were determined by two-way ANOVA for repeated measures followed by Dunnett’s multiple comparison test comparing all group means against non-transduced control group. Overall P-value < 0.001 for a and b; ** Adjusted-P ≤ 0.01, *** Adjusted-P ≤ 0.001, **** Adjusted-P ≤ 0.0001. c Representative plots showing phosphorylation of CD3ζ after 5 days expansion in the presence of IL-2 and anti-CD3/anti-CD28-coupled Dynabeads. CD3ζ phosphorylation in GFP-positive T cells was analyzed by flow cytometry. Graph shows representative data of two independent experiments

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