Figure 5.

Immunization with ileum content modifies intestinal immunity and ileum microbiota. C57Bl/6 mice were immunized s.c. with 200 μl of either PBS or the ileum contents of mice fed HFD (treated). Thirty-five days later, mice were fed either NC or HFD as illustrated in (Figure 1A). (A) After one month of HFD, SILP leukocytes (CD4 + TCRβ + CD45+ cells) from mice fed NC, HFD or HFD treated mice were analyzed by flow cytometry. Proportion of IFNγ, TNFα and IL17-producing CD4 T cells upon stimulation and Foxp3 CD4 T cells. Graphs show mean ± SEM; n = 5 mice per group, one experiment representative of two. Data with similar superscript letters (a,b) indicate data non-statistically different between each other at a probability threshold p > 0.05. (B) After one month of HFD, IgA concentration was measured in cecum contents from mice fed NC, HFD or from HFD treated mice, by ELISA. Graphs show mean ± SEM; n = 5 mice per group. Data with similar superscript letters (a,b) indicate data non-statistically different between each other at a probability threshold p > 0.05. (C–E) Quantification and characterization of the microbial composition of ileum mucosa of NC, HFD and HFD treated mice. (C) 16S rDNA concentration. (D) 16S rRNA-DNA gene sequencing analysis of the ileum mucosa microbiota. Principal Coordinate Analysis (PCoA); (E) Frequency of major bacterial genera in NC, HFD or HFD treated mice obtained from ileum mucosa 16s rRNA-DNA gene sequencing analysis. Graphs show mean ± SEM; n = 5 mice/group; Data with similar superscript letters (a,b) indicate data non-statistically different between each other at a probability threshold p > 0.05.