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Purpose: Specific oral immunotherapy (SOIT) using interferon-γ (IFN-γ) has been successful as a food allergy treatment. Interleukin-10 (IL-10)-producingregulatory B cells (Br1s) play a role in immune tolerance to food allergens. In addition, IFN-γ shows tolerogenic effects on allergen-inducedBr1 responses. Methods: Eleven patients that were allergic to cow’s milk and 12 milk-tolerant subjects were selected by double-blind placebocontrolledfood challenge (DBPCFC) and clinical characteristics. The immunomodulatory effects of IFN-γ on allergen-specific Br1 responses wereevaluated in 6 milk allergy patients and 8 milk-tolerant subjects. Peripheral blood mononuclear cells (PBMCs) from subjects were stimulated withcasein and/or IFN-γ and analyzed by flow cytometry. Results: IFN-γ had no effect on total cell counts or the proportion of Br1 cells in PBMCs. IFN-γstimulation did not change total Br1 cell counts or the percentage of Br1s among CD5(+) B cells in the milk allergy or the milk-tolerant groups. In themilk allergy group, Br1 counts were not different between the control and the casein stimulation but significantly increased in the IFN-γ + caseinstimulated cells, and the Br1 fractions were decreased after casein stimulation and recovered in the addition of IFN-γ for stimulation. In the milk-tolerantgroup, Br1 counts increased in the casein stimulated cells and in the IFN-γ + casein stimulated cells, but the increase was significantly lesswhen IFN-γ was added, and the Br1 fractions were increased after casein stimulation and IFN-γ + casein stimulation, that was not significant whenIFN-γ was added. Conclusions: IFN-γ-induced allergen-specific Br1 responses in the PBMCs of milk allergy patients play a role in milk allergenspecifictolerance induction in vitro. Further investigations into the molecular immunological mechanisms underlying the induction of allergen-specificBr1 responses are needed.