Article Figures & Data
Figures
- Figure 1.
Microbial community composition in tumor-free or mice bearing B16-F10 melanoma or LLC with or without dysbiosis. Microbial composition in healthy controls (orthobiotic, Ortho) and antibiotic exposed (dysbiotic, Dys) mice with and without B16-F10 melanoma or LLC at the order (A) and family (B) levels. C, Log2 abundance heatmap of microbial families. The relative abundance for the microbial families is indicated by hue. D, Taxonomic cladograms obtained from LEfSe analysis of 16S rRNA sequences (blue, no tumor; red, B16-F10; green, LLC). Yellow circles represent nonsignificant differences in abundance between the groups. E, LEfSe analysis revealing differentially enriched microbial groups across tumor types (blue, no tumor; red, B16-F10; green, LLC) within each of the orthobiotic and dysbiotic groups. For A–C, independent samples (n = 4 each) were merged (i.e., ASV read counts summed) into their respective groups prior to visualization. The letters (a)–(i) in D refer to the microbial groups listed in E.
- Figure 2.
Dysbiosis alters the gastrointestinal tract and enhances melanoma and lung carcinoma progression. The small intestines (A), ceca (B), and colons (C) of mice bearing B16-F10 melanoma or LLC cells with or without antibiotic-induced dysbiosis. Representative hematoxylin and eosin images of small intestines (D) and colons (E) in B16-F10 melanoma–bearing mice. Growth curves and masses of B16-F10 melanoma (F) and LLC (G). Tumor growth curves are shown in mean volumes ± SEM with nonlinear regression-fit lines and tumor weights are shown as mean mass ± SEM (n = 10–18 animals per group pooled from two or three individual experiments). Dysbiosis was induced by exposing mice to antibiotics (ampicillin, neomycin, metronidazole, each at 250 mg/L; vancomycin at 125 mg/L). Scale bar, 200 μm. *, P < 0.05, **, P < 0.01, ***, P < 0.05, two-sided t test; ns, not significant.
, orthobiotic (Ortho); , dysbiotic (Dys) mice. - Figure 3.
Tumor vascular adhesion molecules are suppressed under dysbiotic conditions. A, Representative analysis plots for ICAM-1 (CD54) on TECs (CD45− CD31+). Quantification in percentage and absolute number of ICAM-1 (CD54; B), VCAM-1 (CD106; C), MCAM (CD146; D), E-selectin (CD62E; E), and P-selectin (CD62P) expressing TECs (F). Data are the mean ± SEM (n = 5–11 B16-F10 tumors per group, pooled from 2–3 individual FACS experiments). *, P < 0.05, two-sided t test; ns, not significant.
, orthobiotic (Ortho); , dysbiotic (Dys) mice. - Figure 4.
Dysbiosis decreases the abundance of activated and effector T cells in tumors. B16-F10–infiltrated CD3+ CD8+ T cells (A) and CD3+ CD4+ T cells (B) with or without dysbiotic conditions. Percentage and amount of CD3+ CD8+ T cells expressing CD44 (C), CD69 (D), CD27 (E), and CCR7 (F). Data presented as means ± SEM and is representative of two independent experiments n = 4–5 per group. *, P < 0.05; ***, P < 0.001, two-sided t test; ns, not significant.
, orthobiotic (Ortho); , dysbiotic (Dys) mice. - Figure 5.
TNFα is suppressed under dysbiotic conditions. Percentage and amount of TNFα-producing (A) or IFNγ-producing (B) tumor-infiltrating CD3+ CD8+ CD44+ T cells and systemic TNFα serum levels (C) under orthobiotic and dysbiotic conditions. Data presented as means ± SEM (n = 4–9 C57BL/6 mice bearing B16-F10 tumors per group pooled from one to three individual experiments). *, P < 0.05; **, P < 0.01; two-sided t test; ns, not significant.
, orthobiotic (Ortho); , dysbiotic (Dys) mice. - Figure 6.
ICAM-1 and T cells are needed for dysbiotic-induced tumor progression and TNFα supplementation rescues T-cell trafficking under dysbiotic conditions. A, Tumor growth curves of B16-F10 melanoma in ICAM-1−/− mice with and without dysbiotic conditions. B, Tumor growth rates of B16-F10 melanoma in immunocompetent (C57BL/6J) and T-cell–impaired Foxn1−/− mice with and without dysbiotic conditions. C, Tumor growth curves of B16-F10 melanoma during dysbiotic conditions with or without TNFα supplementation (every three days with a total of 4 doses; 120 μg/kg). Quantification of ICAM-1 (CD54) expressing CD45− CD31+ TECs (D) and tumor-infiltrated CD3+ CD8+ T cells (E) in B16-F10 tumors during dysbiotic conditions with or without TNFα. Data presented as means ± SEM (n = 4 or 5 B16-F10 tumors per group). *, P < 0.05, two-sided t test; ns, not significant. A, ∇, ICAM-1−/−;
, dysbiotic (Dys) ICAM-1−/−; , wild-type (wt) mice; C, dysbiotic (Dys) mice () and dysbiotic mice () supplemented with TNFα (Dys + TNFα).
Additional Files
Supplementary Data
- Supplemental Figures 1-9 and Table S1 - Supplemental data regarding cecum morphology, body weights, adhesion molecule expression in spleen and thymus, spleen size, tumor and stroma cell viability, and IFN-g and VEGF serum levels after induction of dysbiosis.
Volume 79, Issue 23
