Extended Data Fig. 10: Top-ranked genes from the ORF screen boost antigen-specific T cell responses.
From: A genome-scale screen for synthetic drivers of T cell proliferation
a, b, Co-delivery of anti-CD19 CARs and ORFs to T cells from healthy donors. a, IFNγ and b, IL2 secretion after overnight co-incubation of CD4+ T cells with Nalm6 cells at 1:1 ratio (n = 3 biological replicates, representative of two donors). c, d, IFNγ (c) or IL-2 (d) secretion by CAR+ORF or ORF only T cells co-incubated for 24 h either alone or with Nalm6 cells. e, Cytotoxicity of 19-BBz CAR T cells expressing tNGFR or LTBR ORF after co-incubation with Nalm6 GFP cells. f, Quantification of Nalm6 clearance (relative to Nalm6 co-incubated with untransduced T cells) for CAR+ORF or ORF alone T cells at different effector:target ratios. Unpaired two-sided t-test p values: 0.011, 1.3x10−4, 0.072, 0.02, 0.021, 0.52, 0.087, 1, 0.51 (left to right). g, Representative images of T cells transduced with 19-28z CAR and NGFR or LTBR, co-incubated with CD19+ Nalm6 GFP cells for 48 h at 1:1 ratio. Scale bar: 200 μm. h–j, Repeated stimulation of CAR+ORF T cells with Nalm6 cells. IL-2 secretion (i), or Nalm6 survival (j), by 19-BBz CAR LTBR or tNGFR T cells re-challenged with Nalm6 after repeated stimulation with Nalm6 cells every three days, for up to three rounds of stimulation. k, Secretion of cytokines IL2 and IFNγ by CAR/LTBR or CAR/tNGFR T cells from two patients with DLBCL after overnight incubation with Nalm6 target cells. Two-sided paired t-test p value is shown. l, Representative staining of ORF-transduced T cells endogenously expressing Vγ9Vδ2 TCR. m, Quantification of ORF-transduced T cells expressing Vγ9Vδ2 TCR. n, o, IL2 (n) or IFNγ (o) secretion after 24 h co-incubation of ORF transduced Vγ9Vδ2 T cells with leukaemia cell lines. p, IL2 or IFNγ secretion after 24 h co-incubation of ORF transduced Vγ9Vδ2 T cells with BxPC3, a pancreatic ductal adenocarcinoma cell line. Cell lines in panels n–p were pre-treated with zoledronic acid prior to co-incubation. Error bars indicate SEM. N = 3 biological replicates are shown, unless indicated otherwise.
