Fig. 4. Human CAR T cells armed with NAP exhibit similar advantageous immunological characteristics as murine CAR(NAP) T cells.

a, Self-inactivating (ΔU3) lentiviral constructs used for human T-cell engineering. b,c, Representative histogram (b) and MFI (c) of NAP expression in CAR-positive T cells upon target cell recognition. Dashed line, MFI of isotype staining. d, Relative viability of target cells after exposure to CAR-engineered T cells. e–g, Gene expression in CAR T cells and CAR(NAP) T cells after exposure to target cells. e, Schamatic illustration of the experimental setting. f, GO term enrichment network of the uniquely upregulated genes in CAR(NAP) T cells according to Metascape analysis of the NanoString gene expression data. Numbers in brackets are –log₁₀(P) values. g, Heat map of genes uniquely upregulated in CAR(NAP) T cells presented as normalized z-score of raw gene counts. h, Percentage of DCs migrating towards supernatant collected from a 5:1 co-culture of engineered T cells and Daudi target cells. i–k, Potency of CAR(NAP) T cells assisted by autologous DCs, presented as relative viability of target cells (i), IFN-γ secretion (j) and CD107a expression (k). l–n, CAR-engineered human T cells were co-cultured with autologous immature DCs and Daudi target cells (5:1:1) and assayed for T-cell polarization (l), and production of IL-12 (m) and other cytokines and chemokines (n). All experiments were performed three times and data were pooled. Error bars represent s.e.m. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Precise P values are reported in Supplementary Table 3.