Extended Data Fig. 8: Efficacy of 11-mix in enhancing treatment of MC38 tumours in the context of a complex microbiota.
From: A defined commensal consortium elicits CD8 T cells and anti-cancer immunity
a, Experimental design for SPF mouse-based studies in b and c (and in Fig. 4d–k). SPF mice were subjected to treatment with AVMN (from day −7 to day 2) and subcutaneous implantation of MC38 adenocarcinoma or BrafV600E Pten−/− melanoma cells on day 0. The mice were reconstituted with SPFfae on day 3. For the 11- or 10-mix treatment groups, the initial oral administration was done simultaneously with SPFfae on day 3, followed by repetitive dosing of the 11- or 10-mix alone, two or three times per week until the end of the experiment. An anti-PD-1 or anti-CTLA-4 antibody was injected intraperitoneally every third day between days 3 and 9. b, Representative photograph of excised MC38 tumours on day 23. Scale bar, 10 mm. c, H&E staining, along with histology score, of the colon on day 27 (SPF, SPF+anti-PD-1, SPF+11mix, and SPF+11mix+anti-PD-1 groups) and on day 44 (SPF+anti-CTLA-4 and SPF+11mix+anti-CTLA-4 groups). Scale bar, 50 μm. For comparison, the colonic histology score of SPF Il10−/− (colitis-prone) mice is shown. d, Experimental design for GF+donor C human microbiota-based studies in e and f. C57BL/6 germ-free mice were colonized with donor C human faecal microbiota or left uncolonized on day 0. For the 11- or 10-mix treatment groups, the initial oral administration was done simultaneously with the donor C human faecal sample on day 0, followed by repetitive dosing of the 11- or 10-mix alone two or three times per week until the end of the experiment. e, Faeces were collected at the indicated time points. The relative abundance of each of the 11 strains’ DNA was determined by qPCR. Colonization with all 11 strains was confirmed. f, Percentage of IFNγ+ CD8 T cells in the colonic lamina propria and iLNs at day 21 was enumerated by flow cytometry. g, h, C57BL/6 germ-free mice were inoculated with donor C faecal samples with or without 11- or 10-mix, following the same protocol as in d. Mice were then subjected to subcutaneous implantation of MC38 cells on day 0. Anti-PD-1 was injected intraperitoneally every third day between days 3 and 9. Experimental design is shown in g and tumour growth data of MC38 is shown in h. Each circle represents an individual animal, except in h, where each circle represents the mean. The number of mice in each group is shown. Red, grey and brown asterisks show significance versus the C+11-mix+anti-PD-1, C+anti-PD-1, and C+10-mix+anti-PD-1 groups, respectively. Data are mean and s.e.m. (e) or s.d. (all others). ***P < 0.001; **P < 0.01; *P < 0.05; one-way (f) or two-way (h) ANOVA with Tukey’s test. See Source Data for exact P values.
