Table 1.
Primers Used for RT-qPCR.
Fig 1.
Infection with ME49Δα-amy tachyzoites significantly mitigated systemic inflammatory responses induced by T. gondii tachyzoites.
A. H&E staining of the mice heart, lung, liver, spleen and colon tissue samples (n = 3 per group). B-F. Histological scores of heart, lung, liver, spleen and colon tissues. G-L. The mRNA expression levels of TNF-α, IL-1β, IL-6, IL-12, IL-10, and TGF-β were measured by qRT-PCR (n = 3 per group). The Data are presented as the mean ± SEM of three independent experiments. Statistical significance was done with the Student’s t-test. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.
Fig 2.
Infection with the ME49Δα-amy strain significantly alleviated T. gondii-induced intestinal barrier damage.
The expression levels of occludin and ZO-1 were measured by immunofluorescence. Occludin protein was labeled with a green fluorophore, ZO-1 protein was labeled with a red fluorophore, and the cell nucleus was labeled with a blue fluorophore (n = 3 per group). The Data are presented as the mean ± SEM of three independent experiments. Statistical significance was done with the Student’s t-test. ****p < 0.0001.
Fig 3.
Differential gut microbiota profiles in mice following infection with different virulence tachyzoites.
A. Microbial community diversity (measured by Shannon index). B. Microbial community abundance (measured by Chao1 index). C. Principal coordinate analysis (PCoA) based on the weighted UniFrac distance matrix. D. Relative abundances of fecal bacterial at phylum level. E. Relative abundances of fecal bacterial at family level. F. Relative abundances of fecal bacterial at genus level. G. Heat map of the top 50 microbiota at genus level in fecal samples from mice infected with ME49 or ME49Δα-amy tachyzoites. H. Differentially abundant taxa of fecal microbiota was analyzed by LEfSe. LDA score ≥ 2. I. KEGG pathway analysis of differentially expressed genes. J. Heatmap of Spearman’s correlation between gut microbiota abundance and inflammatory factors. *p < 0.05, **p < 0.01.
Fig 4.
Changes in the fecal metabolomic profiles among different treatment groups.
A. Total metabolome classifications of compounds with differential metabolites. B-C. Fecal metabolome profiles were clustered using PLS-DA. D. Volcanic map analysis of differential metabolites. E. Metabolite relative abundances were clustered by UPGMA and visualized in a heatmap using Z-scores. F-H. The concentrations of fecal Fructose, Glucose, and N-Acetyl-D-glucosamine were displayed as box and dot plots. I. SMPDB pathway enrichment analysis according to the markedly altered metabolites. J. The concentration of GlcNAc was measured in serum. Four independent biological replicates were analyzed for each group. Statistical significance was done with the Student’s t-test. **p < 0.01.
Fig 5.
Interrelationships between the gut microbiota and the metabolome were assessed by Spearman’s correlation analysis.
The heatmap displays correlation coefficients between differentially enriched bacterial taxa and metabolites in response to ME49Δα-amy tachyzoites infection. Red represents positive correlation, and blue represents negative correlation. ***p < 0.001, **p < 0.01, *p < 0.05.
Fig 6.
Inhibitory effect of GlcNAc on T. gondii-induced inflammation in RAW264.7 cells.
The mRNA expression levels of TNF-α (A), IL-1β (B), IL-6 (C), IL-12 (D), IL-10 (E), and TGF-β (F) were measured by qRT-PCR. The Data are presented as the mean ± SEM of three independent experiments. Statistical significance was done with the one-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.
Fig 7.
Oral administration with GlcNAc alleviated T. gondii-induced systemic inflammation in mice.
A. Body weight changes of mice (n = 10 per group). B. The parasite loads in peritoneal fluids were determined by quantitative PCR (n = 3 per group). C. H&E staining of the mice heart, lung, liver, spleen and colon tissue samples. D-H. Histological scores of heart, lung, liver, spleen and colon tissues (n = 3 per group). I-N. The mRNA expression levels of TNF-α, IL-1β, IL-6, IL-12, IL-10, and TGF-β were measured by qRT-PCR (n = 3 per group). The Data are presented as the mean ± SEM of three independent experiments. Statistical significance was done with the one-way ANOVA. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.
Fig 8.
Oral administration with GlcNAc significantly alleviated T. gondii-induced intestinal barrier damage.
The expression levels of occludin and ZO-1 were measured by immunofluorescence. Occludin protein was labeled with a green fluorophore, ZO-1 protein was labeled with a red fluorophore, and the cell nucleus was labeled with a blue fluorophore (n = 3 per group). The Data are presented as the mean ± SEM of three independent experiments. Statistical significance was done with the one-way ANOVA. ****p < 0.0001.