Figure 1.
Biopsies taken pre-challenge (week 20) and post-challenge (week 21) from trial 1 and were cultured for 24 hours in tissue culture medium, then supernatants taken for CBA analysis of IFN-γ (A) and IL-17A (B).
One-way ANOVA was carried out comparing between groups and timepoints. Unless otherwise indicated differences are not significant. ** = p<0.01, * = p<0.05.
Figure 2.
PBMCs from trial 1 pre-challenge (week 20) and post-challenge (week 21) were prepared, cryopreserved and thawed together, then stained for CD4, Foxp3 and CD25.
Proportions of CD4+ cells in the PBMC lymphocyte population (A). Proportions of CD25+Foxp3+ cells in CD4+ population (B). CD25 MFI (C) and Foxp3 MFI (D), both in the CD4+CD25+Foxp3+ population. Representative Foxp3 versus CD25 plots of cells gated on CD4+ PBMCs are shown in (E). Cell suspensions were prepared from duodenal biopsies pre-challenge and post-challenge, and stained for CD4 and Foxp3. Foxp3+ proportion of the CD4+ population of biopsy cells (F). Formalin fixed duodenal biopsies were also stained for Foxp3 by immunohistochemistry, Foxp3+ cells per high-power field of view (G). Where paired data was available (all except (F)), data were analysed by two-way ANOVA, and if a significant interaction was found paired t tests were used to show differences within groups. For (F), data were analysed by non-parametric ANOVA. N.S. = not significant, * = p<0.05, ** = p<0.01.
Figure 3.
Duodenal biopsies were taken from trial 2 participants pre-infection (Pre-Inf, week 0), pre-challenge (Pre-Ch, week 20) and post-challenge (Post-Ch, week 21) and cultured in either medium alone or with 50 µg/ml QE65 peptide for 24 hours at 37°C in 95% O2/5% CO2.
Supernatants were then taken and levels of cytokines measured by CBA. Results shown in A–C are from the pre-infection timepoint only, showing both medium and QE65 levels. All other results (D–I) have had medium controls subtracted. Cytokines shown are IL-2 (A, D), IFN-γ (B, E), IL-17A (C, F), IL-10 (G), IL-5 (H) and IL-13 (I). A–C was analysed by paired t tests, D–I by 1-way ANOVA. Where data was not normally distributed, log-transformation was carried out and parametric tests used. Differences are not significant unless indicated. * = p<0.05, ** = p<0.01.