Forkhead box protein 3 demethylation is associated with tolerance induction in peanut-induced intestinal allergy

Background: Regulatory T (Treg) cells play an essential role in the maintenance of immune homeostasis in allergic diseases. Objectives: We sought to define the mechanisms underlying induction of tolerance to peanut protein and prevention of the development of peanut allergy. Methods: High or low doses of peanut extract were administered to pups every day for 2 weeks before peanut sensitization and challenge. After challenge, symptoms, Treg cell numbers, and forkhead box protein 3 (Foxp3), T(H)2 and T(H)17 cytokine, and Tgf beta expression in mesenteric lymph node (MLN) CD4(+) T cells and jejunum were monitored. Treg cell suppressive activity and Foxp3 methylation in MLN CD4(+) T cells were assayed. Results: Feeding high but not low doses of peanut before sensitization induced tolerance, as demonstrated by prevention of diarrhea and peanut-specific IgE responses, increases in the percentage of CD4(+)CD25(+)FoxP3(+) cells in MLNs, and Foxp3 mRNA and protein expression in CD4(+) cells from MLNs or jejunum. Feeding high doses of peanut before sensitization decreased percentages of CD3(+)CD4(+)IL-13(+) and CD3(+)CD4(+)IL-17(+) cells in MLNs and decreased Il13 and Il17a and increased Tgf beta mRNA expression in the jejunum; numbers of CD103(+) dendritic cells in MLNs were significantly increased. Treg cell suppression was shown to be antigen specific. Foxp3 methylation was increased in peanut extract-sensitized and challenged mice, whereas in tolerized mice levels were significantly reduced. Conclusions: Feeding high doses of peanut to pups induced tolerance to peanut protein. Foxp3 demethylation was associated with tolerance induction, indicating that Treg cells play an important role in the regulation of peanut sensitivity and maintenance of immune homeostasis.