Extended Data Fig. 5: Anti-tumour efficacy, tumour infiltration, phenotype and function of o2R and o9R pmel T cells.
From: Potentiating adoptive cell therapy using synthetic IL-9 receptors
a,b, Survival of B16-F10 tumour-bearing mice treated with (a) o2R or (b) o9R pmel T cells and MSA-IL2 or MSA-oIL2. BL/6 T cells treated with MSA-IL2 were used as an off-target T cell control. Untransduced pmel T cells plus mIL-2 in tumour-bearing lymphodepleted and non-lymphodepleted mice served as controls. c, Effect of o9R pmel T cells in a lymphodepleted host. Tumour growth (mean ±SEM, left panel) of B16-F10 tumours in lymphodepleted C57BL/6 mice treated with o9R pmel T cells and MSA-IL2, MSA-oIL2, or no IL2. d, Individual B16-F10 tumour growth curves related to survival curves shown in Fig. 2e. All mice were non-lymphodepleted except one group (in purple) which received total body irradiation (5Gy). CR, complete regression. *, p < 0.05; **p < 0.01; ****, p < 0.0001 (ANOVA). e, opt-SNE clustering of CD45+ tumour-infiltrating leukocytes seven days after adoptive transfer of o2R (left) and o9R (middle) pmel T cells (n = 4 mice/group; mice were treated without lymphodepletion and treated MSA-oIL2 2.5x104 units/day for five days starting with ACT). Volcano plot of differentially abundant clusters in tumours from mice treated with o9R versus o2R pmel T cells (right panel). f, opt-SNE clustering of the subset of tumour infiltrating o9R and o2R pmel T cells in non-lymphodepleted hosts treated with MSA-oIL2 (n = 4 mice/group, left panel), with separate plots for each treatment group illustrating only differentially abundant clusters (middle panel), and a volcano plot of differentially abundant clusters annotated with distinguishing features (right panel). g, Tumour-infiltration of CD3+CD8+ T cells and CD8+PD1+ T cells in non-lymphodepleted hosts treated with either o2R and MSA-oIL2 or o9R pmel T cells and MSA-oIL2 by multiplex IHC (red = CD3, orange = CD8, yellow = PD1, green = CD4, teal = FOXP3). Images are representative of tumours from n = 3 mice/group and of one independent experiment (conclusions verified in three independent experiments by flow and mass cytometry). Quantifications (mean ± SD) shown to the right (n = 3 biological replicates/group). h, In vitro growth (mean ± SD, n = 3 biological replicates/group) of nRFP+ B16-F10 tumour cells cocultured with o2R or o9R pmel T cells (2:1 E:T ratio) pretreated with MSA-IL2 (50nM). i, IFNγ secretion by oIL-2R and oIL-9R pmel T cells cocultured with B16-F10 melanoma in vitro for 24 h. T cells were pretreated with MSA-oIL2 (5 µM) for 48 h in vitro prior to coculture (mean ± SD, n = 3 biological replicates/group). *, p < 0.05; unpaired t-test, two-sided.
