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Background  Epicutaneous immunotherapy ( EPIT ) is a promising method for treating food allergies. In animal models,  EPIT  induces sustained unresponsiveness and prevents further sensitization mediated by Tregs. Here, we elucidate the mechanisms underlying the therapeutic effect of  EPIT , by characterizing the kinetics of  DNA  methylation changes in sorted cells from spleen and blood and by evaluating its persistence and bystander effect compared to oral immunotherapy ( OIT ).    Methods   BALB /c mice orally sensitized to peanut proteins ( PPE ) were treated by  EPIT  using a  PPE ‐patch or by  PPE ‐ OIT . Another set of peanut‐sensitized mice treated by  EPIT  or  OIT  were sacrificed following a protocol of sensitization to  OVA .  DNA  methylation was analyzed during immunotherapy and 8 weeks after the end of treatment in sorted cells from spleen and blood by pyrosequencing. Humoral and cellular responses were measured during and after immunotherapy.    Results  Analyses showed a significant hypermethylation of the  Gata3  promoter detectable only in Th2 cells for  EPIT  from the 4th week and a significant hypomethylation of the  Foxp3  promoter in  CD 62L +  Tregs, which was sustained only for  EPIT . In addition, mice treated with  EPIT  were protected from subsequent sensitization and maintained the epigenetic signature characteristic for  EPIT .    Conclusions  Our study demonstrates that  EPIT  leads to a unique and stable epigenetic signature in specific T‐cell compartments with downregulation of Th2 key regulators and upregulation of Treg transcription factors, likely explaining the sustainability of protection and the observed bystander effect.

allergy

Gata3  hypermethylation and  Foxp3  hypomethylation are associated with sustained protection and bystander effect following epicutaneous immunotherapy in peanut‐sensitized mice