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Summary   Background  Allergen‐specific immunotherapy favours immune deviation from a Th2 to a Th1 response and increases the number of regulatory  T  cells (Tregs). Epicutaneous immunotherapy ( EPIT ) of sensitized mice decreases the clinical and the allergen‐specific Th2 responses and increases local and peripheral  F oxp3 +   T regs.    Objective  To investigate the role of  T regs in  EPIT  and characterize their phenotype and maintenance following  EPIT .    Methods  Tregs were investigated using  in vivo  depletion or adoptive transfer into  BALB /c mice. Tregs were depleted using anti‐ CD 25 antibody injection during  EPIT , and allergen‐specific responses were compared with Sham,  EPIT  alone and naïve mice. To demonstrate that Tregs can mediate protection by their own, and to study their maintenance following the end of  EPIT ,  CD 25 + CD4 +  Tregs isolated from mice just after or 8 weeks after EPIT were transferred into peanut‐sensitized mice. Foxp3‐ IRES ‐ mRFP  mice were transferred with  EPIT ‐induced Tregs to analyse the induction of host  T regs.    Results  The anti‐ CD 25 antibody injection to  EPIT  mice abrogated the induction of  T regs in spleen and the expression of  F oxp3 in oesophagus. This resulted in levels of peanut‐induced eosinophilic infiltration in oesophagus similar to  S ham and significantly higher than  EPIT . Whereas the transfer of  T regs from Sham‐treated mice demonstrated no effect, the transfer of  T regs isolated just after  EPIT  prevented peanut‐induced eosinophil infiltration and eotaxin expression and induced  F oxp3 in oesophagus. The transfer of  T regs isolated 8 weeks after  EPIT  suppressed allergen‐specific responses as efficiently as did Tregs isolated just after  EPIT  and increased spleen Foxp3 +   CD 25 +   CD 4 +  cells similarly. The use of reporter mice demonstrated an increase in host  T regs.    Conclusions  These results confirm the  T regs‐mediated mechanism of  EPIT  and demonstrate the persistence of efficient  T regs during a long period of time after treatment cessation. This suggests that  EPIT  induces long‐term tolerance in peanut‐sensitized mice.

clinical and experimental allergy

The regulatory  T  cells induction by epicutaneous immunotherapy is sustained and mediates long‐term protection from eosinophilic disorders in peanut‐sensitized mice