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. 2020 Apr 22;11:1922. doi: 10.1038/s41467-020-15857-x

Fig. 2. Trehalose derived from Hp induces CD8+ Treg cells.

Fig. 2

a Forty-eight metabolites identified in the contents of the small intestines of five uninfected and four Hp-infected mice (a), and 33 metabolites in DMEM used to culture adult worms [Hp excretory/secretory (HES) antigens] (d) using GC/MS were subjected to univalent analyses, and volcano plots are shown. Trehalose in the small intestines of b and trehalose concentrations among HES antigens (e) were measured by GC/MS. c Trehalose concentrations in serum from the indicated mice were measured using ELISA. f Splenic CD8+ Treg cells in mice orally administered HES antigens or HES antigens exposed to trehalase and mice infected with Hp were analysed as described in Fig. 1d, and the frequency is shown. g Blood glucose concentrations were monitored in these mice after T1D induction. h–j Mice fed trehalose (TH) and maltose (MT) as a control disaccharide were subjected to T1D induction. Hp-infected (Hp) and uninfected (DW) mice were used as positive and negative controls, respectively. Splenic CD8+ Treg cells were quantified 2 weeks after feeding as described in Fig. 1d, and the percentages of these cells are shown (h). Blood glucose (i) and plasma insulin levels (j) were analysed as described in Fig. 1a, b. k Female NOD mice were fed with TH after hyperglycaemic onset. l Glucose levels of individual mice with hyperglycaemia of <350 mg/dl (left panel) or more than 350 mg/dl (right panel) at the beginning of sugar feeding. m Splenic CD8+ Treg cells in NOD mice used in l were quantified at 2 weeks after feeding trehalose as described in Fig. 1d, and the percentages of these cells are shown. Values represent the mean ± S.D. of 10 mice except for five mice in b. Asterisks denote statistical significance at p < 0.05 calculated by the two-tailed Mann–Whitney test (b, e), two-sided unpaired Student’s t-test (c), two-way ANOVA (g, i, k) and Tukey post-hoc analysis (f, h, i, m). NS indicates not significant. All experiments except for GC/MS run (a) and NOD mice (m, l) each once, were repeated at least three times with similar results.