Fig. 6: Effect of antibiotic-induced microbiota alterations on placental metabolome.

PCA of placental metabolome in vancomycin-treated (green) and control (blue) dams at ed14.5 (n = 6, a). Pathway (b) and Metabolite Set Enrichment Analysis (c) of placental metabolites with a VIP score >1.5 at ed14.5. Global test algorithm was employed for both pathway impact and pathway enrichment analysis. Box and Whisker plot representation of galactose, and galactitol placental relative abundance at ed14.5 (n = 6 experiments, d). The plot shows all individual data points. Horizontal lines within the plot indicate the median values. Placental NK cells of control dams (ed14.5) significantly reduced IFN-γ production after 5 hours stimulation with PMA/ionomycin in media supplemented with a glucose:galactose ratio of 1:3 (n = 5 experiments, e, p = 0.0209). Placental NK cells of vancomycin-treated dams orally treated with 30 g/kg of glucose before sacrifice (ed14.5) show increased IFN-γ (p = 0.0151) production and less TNF (p = 0.0440) production after 5 hours stimulation with PMA/ionomycin in media (filled squares) compared to vancomycin-treated dams (filled circles) (f). Two-sided Unpaired t test (e), the plot shows all individual data points. Horizontal lines within the plot indicate the median values. Lines represent mean plus SEM, two-sided Unpaired t test (n = 4 experiments, f), *p < 0.05, ****p < 0.0001. Glucose administration to vancomycin-treated dams (n = 9, 30 g/kg per 3 days before sacrifice at ed14.5, squared filled symbols) decreases pregnancy loss compared to vancomycin treatment alone (n = 10 experiments, round filled symbols, g, p = 0.0011). Lines represent mean plus SEM, two-sided Mann and Whitney test, **p < 0.01. Each measurement was taken from a single experiment. See also Supplementary Figs. 12–14. Source data are provided as a Source Data file.