Fig 2: Neutralization of IL-17A abolishes the worse outcome of Il10−/− mice. Triphenyltetrazolium chloride staining was used for evaluation of infarct volume at day 7 of IL-17A antibody (A) or IgG control (C) treated Il10−/− and WT control mice. Bederson score was performed 7 days after tMCAO IL-17A antibody (B) or in IgG control (D) treated group. Infarct data are presented as mean ± SEM of 9–11 WT and 6–9 Il10−/− mice per group. Bederson score as mean ± SEM of 9–11 WT and 7–10 Il10−/− mice per group. Statistical significances were analyzed by Student t test (A, C) and Mann–Whitney U test (B, D). *P < 0.05

Fig 3: Clinical and immunological impact of the adoptive transfer of trained-macrophages on A20 cells B lymphoma tumor growth in mice. (A) Schematic representation of the in vivo induction of B lymphoma in mice and scheduling of trained macrophage infusion. Mice (n = 7 in each group) were subcutaneously injected with 2 × 106 A20 cells and sacrificed at day 34 in one experiment. (B) Tumor volume evolution in the different groups of A20-challenged mice (n = 7). Tumor growth was monitored three times a week with a microcaliper, and tumor volumes were calculated as length × width2 × 0.5236 and expressed in mm3. Statistics are shown between the BCG-macrophages infused (A20-BCG) and the untreated A20 groups. Differences were detected at sacrifice using the Mann–Whitney test. **p ≤ 0.01. (C) Cytokine production in the supernatant of splenic cells harvested at sacrifice, stimulated with 5 μg/ml of Concanavalin A and incubated for 48 h at 37 °C with 5% CO2. Levels of TNF-α, IL-6, and IL-10 were assessed by ELISA and concentrations are expressed in pg/ml. Data represent the mean ± SEM from the duplicate of at least n = 5 biologically independent samples. (D, E) Flow cytometric analysis of splenic and tumoral CD4+ (C) and CD8+ (D) T-cell phenotype at day 34 of the experiment. Tumor infiltrating T cells were gated on CD3 and CD4 or CD8 positive cells among CD45 positive population. Data represent the mean fluorescence intensity of CD69 ± SEM, reflecting the activation status of the assessed cells, obtained from at least n = 5 biologically independent samples. (F, G) RT-qPCR assessment of fizz1, inos, il10, tgfb, foxp3, il4, il6, and infg mRNA levels in the spleen and tumor lesions of the different groups of A20-challenged mice at day 36 of the experiment. Results were expressed as fold increase versus the untreated A20-mice group, derived from at least n=5 biologically independent samples. Unpaired T-test was used to detect the differences between the groups. NS, non-significant; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001.

Fig 4: Murine fecal metabolites suppress activated immune cells and are enriched with hypoxanthine/inosine and glutamate/glutamine.A Schematic illustration of in vitro fecal supernatant–immune cell cultures and stool metabolite analysis. mRNA expression for Il10 (B), Foxp3 (C), and Tnfa (D) as a ratio to Gapdh in RRV-primed hepatic MNCs cultured in the presence of fecal supernatants from neonatal mice of water- or butyrate-fed mothers (mean ± SD, two-tailed unpaired Student’s t test, n = 4 per group; *p < 0.05, **p < 0.01, ***p < 0.001). E NMDS and ANOSIM of metabolites in fecal supernatants of neonatal mice from water- or butyrate-fed mothers at 14 days of age. F Volcano plot illustrating fecal metabolites with p values and fold changes between neonatal mice from water- and butyrate-fed mothers (inset depicts metabolites of lower distance). The replicate values were determined using biologically distinct samples and p values calculated using unpaired Student’s t test with two-tailed distribution. G Fecal metabolites ordered by Euclidean distance measured from the volcano plot. Source data for this figure are provided as a Source data file.

Fig 5: Treatment of neonates with butyrate decreases hepatobiliary injury.A Diagrammatic outline of gavaging RRV-infected neonatal mice with sodium butyrate. B Jaundice (generalized linear mixed effect model with logit link and two-sided Wald test with Bonferroni correction; ****p < 0.0001) and C survival (two-sided log-rank test; ***p < 0.001) rates in RRV-infected newborn mice treated daily with butyrate or PBS. D Virus titers in EHBD and livers at day 7 after RRV infection of newborn mice from water-fed dams (mean ± SD, two-tailed unpaired t test with Welch’s correction; n = 5 per group; ns = not significant) and E section of EHBD from butyrate-treated mice 14 days after RRV. In all, 15–30 EHBD sections (corresponding to >100 sections at ×200 or ×400 magnification fields from n = 11 mice) stained with H&E per tissue specimen were evaluated for histology analysis. Scale bar = 50 µM. Foxp3 (F) and Il10 (G) mRNA in RRV-naive or primed hepatic mononuclear cells cultured with or without butyrate, normalized to Gapdh (mean ± SD, two-tailed ANOVA with Duncan’s multiple comparisons, n = 3 per group. *p < 0.05, **p < 0.01, ns = not significant). Source data for this figure are provided as a Source data file.