Extended Data Fig. 2: KPP STAMP microtumour rejection is mediated by tumour antigen specific CD8 T cells.
From: In situ tumour arrays reveal early environmental control of cancer immunity
a, Automated analysis of growth kinetics of KPP-EGFP total tumour array area (mm2) per animal in Rag-2-deficient and wild type animals as described in Fig. 1b, n = 5 animals per group. b, Representative images of KPP-EGFP tumour arrays in wild type (upper panel) or Rag-2-deficient (lower panel) mice at 6 and 14 days post tumour implantation related to b. Red encircled tumours are rejected and white encircled tumours are persistent between time points. c, Survival probability of individual tumours of KPP-EGFP arrays in mice treated with CD8 depleting (n = 318 tumours, 5 animals) or isotype control antibodies (n = 534 tumours, 5 animals). d, Automated analysis of growth kinetics of individual tumour area (mm2) for experiment as described in Fig. 1d. Red lines indicate tumours that are rejected, grey lines indicate tumours that persist. n = 100 tumours, n = 5 animals in Rag-2-deficient animals reconstituted with WT T cells, n = 88 tumours, n = 5 animals in Rag-2-deficient animals reconstituted with OT-I T cells. e, Representative images of STAMP tumour arrays for experiment as described in d. Red = T cells, green = KPP-EGFP. f, T cell infiltration kinetics of individual tumours measured by tdTomato MFI of T cells for experiment as described in d. n = 5 animals per group, n = 61 tumours in Rag-2-deficient animals reconstituted with WT T cells and n = 53 tumours in Rag-2-deficient animals reconstituted with OT-I T cells. Data are mean +/− s.e.m. Statistical analysis was performed using a two-tailed Mann-Whitney U-test. g, Representative image time course of KPP-OVA-EGFP tumour arrays in Rag-2-deficient animals reconstituted with wild type or OT-I T cells. n = 4 animals reconstituted with WT T cells, n = 5 animals reconstituted with OT-I T cells, n = 156 tumours in animals reconstituted with WT T cells, n = 93 tumours in animals reconstituted with OT-I T cells. Red = T cells, green=KPP-EGFP. h, Survival probability of individual tumours from mice bearing KPP-EGFP-OVA tumour in Rag-2-deficient animals reconstituted with wild type or OT-I T cells and treated with CD8 depleting or isotype control antibody. n = 4 WT-OVA isotype, n = 5 OT-I-OVA isotype and n = 5 OT-I-OVA anti-CD8 treated animals per group, n = 93 OT-I-OVA isotype, n = 156 WT-OVA isotype and n = 206 OT-I-OVA anti-CD8 treated tumours. Statistical analysis was performed with log-rank test (referenced to WT-OVA isotype). i, Frequency of tumour antigen (M86) specific T cells in M86-encoding RNA-LPX vaccinated or naive T cell donor mice shown as PD-1+ percent of CD8+ activated memory T cells. j, Survival probability of individual tumours from mice bearing KPP-M86-mTagBFP2 microtumours in Rag-2-deficient mice reconstituted with tumour antigen specific T cells (M86) from vaccinated mice or T cells from naive mice. n = 73 tumours in vaccinated animals, n = 72 tumours in naive animals, 4 animals per group pooled. c,f,h,i, P values are shown in the graph and colour-coded for the treatment group in h. c,h,j, The centre line shows the Kaplan-Meier curve, the shaded area shows the 95% confidence interval. Statistical analysis was performed with log-rank test.
