Figure 1.
H. p. bakeri-induced intestinal immunopathology.
(A) Enteritis development was evaluated at day 6, 14 and 28 post infection and naïve controls according to a scoring system based on epithelial damage, edema development, villus length, crypt depth and the size of inflammatory cell infiltrates. (B) Goblet cells counts as detected in intestinal cross sections by periodic acid/Schiff stain. Mean + SEM is shown for 5 mice per group; * p < 0.05. Representative cross sections from duodenum (C) and colon (D) stained with H&E (upper row) and periodic acid/Schiff (lower row, goblet cells marked by intense purple stain) for histopathological scoring and goblet cell quantification.
Figure 2.
Cytokine production by mLN cells during acute H. p. bakeri infection.
IL-4 (A), IL-13 (B), IFN-γ (C), IL-10 (D), TGF-β (E) and IL-17A (F) were measured in mLN culture supernatants with (ConA, H.p.b-Ag) or without (-) stimulation. Cells were derived from naïve controls and mice at day 14 post infection. Mean + SEM is shown for 5 mice per group. H.p.b. Ag: H. p. bakeri antigen; Con A: concanavalin A; * p < 0.05; ** p < 0.005.
Figure 3.
Quantification of aerobic intestinal bacterial groups.
Counts of colony forming units (CFU) derived from luminal content of (A) ileum, (B) cecum and (C) colon. Open circles: naïve controls; filled circles: day 14 post H. p. bakeri infection. Mean ± SEM of 5 mice per group is shown. EC: Enterococci, EB: Enterobacteriaceae, LB: Lactobacilli; ** p < 0.005.
Figure 4.
Analysis of 16S rRNA gene-based real-time PCR amplification of intestinal bacterial groups.
Detected DNA levels in luminal content from (A) ileum, (B) cecum and (C) colon of naive (open circles) and acutely infected (day 14 post infection, black circles) mice were calculated as 16S rRNA copy numbers per ng DNA extract. Mean ± SEM is shown for 4-5 mice per group. DB: domain bacteria, γ-p/EB: γ-Proteobacteria/Enterobacteriaceae, LB: Lactobacilli, C IV: Clostridium cluster IV, C XIV a/b: Clostridium cluster XIV a/b, B: Bacteroides group, MIB: mouse intestinal Bacteroides, EC: Enterococci, n.d.: not detectable. * p < 0.05; ** p < 0.005 compared to naïve.
Figure 5.
Proportions of intestinal bacterial group based on 16S rRNA gene real-time PCR.
Pie charts represent the mean frequencies for the main bacterial phyla in (A) ileum, (B) cecum and (C) colon as detected in 4-5 mice per group. Abundance was calculated using the phylum-specific 16S gene copy numbers per ng DNA in relation to the amounts of the DNA copy numbers for the domain Bacteria set to 100%. p.i.: post H. p. bakeri infection.
Figure 6.
Immune response, histopathology and parasite burden of infected wild type versus IL-4Rα-/- mice.
Levels of IL-4 (A), IL-13 (B), IL-10 (C) and IFN-γ (D) produced by mLN cells from naïve controls and mice at day 14 post infection in response to adult worm antigen. (E) Goblet cell counts in duodenal cross sections. (F) Enteritis scores for duodenal tissue. Representative duodenum cross sections from naïve (G) and acutely infected (H) wild type and IL-4Rα-/- mice stained with H&E (upper row) and periodic acid/Schiff (lower row) for histopathological scoring and goblet cell quantification. (I) Adult worm counts of WT and IL-4Rα-/- mice at day 14 post infection. Mean + SEM of 5-6 mice per group is shown. * p < 0.05; ** p < 0.005.
Figure 7.
Quantification of aerobically cultivable bacteria in wild type and IL-4Rα-/- mice.
Counts of colony forming units (CFU) derived from luminal content of (A) ileum, (B) cecum and (C) colon of WT (naïve: open circles; d14 p.i. black circles) and IL-4Rα-/- mice (naïve: open triangles; d14 p.i. black triangles). Mean ± SEM of 5-6 mice per group is shown. CFU: colony forming units, EC: enterococci, EB: Enterobacteriaceae, LB: Lactobacilli; ** p < 0.005.