In Vitro Induction of Allergen-Specific Interleukin-10-Producing Regulatory B Cell Responses by Interferon-gamma in Non-Immunoglobulin E-Mediated Milk Allergy

ABSTRACT

PURPOSE: Specific oral immunotherapy (SOIT) using interferon-gamma (IFN-gamma) has been successful as a food allergy treatment. Interleukin-10 (IL-10)-producing regulatory B cells (Br1s) play a role in immune tolerance to food allergens. In addition, IFN-gamma shows tolerogenic effects on allergen-induced Br1 responses. METHODS: Eleven patients that were allergic to cow's milk and 12 milk-tolerant subjects were selected by double-blind placebo-controlled food challenge (DBPCFC) and clinical characteristics. The immunomodulatory effects of IFN-gamma on allergen-specific Br1 responses were evaluated in 6 milk allergy patients and 8 milk-tolerant subjects. Peripheral blood mononuclear cells (PBMCs) from subjects were stimulated with casein and/or IFN-gamma and analyzed by flow cytometry. RESULTS: IFN-gamma had no effect on total cell counts or the proportion of Br1 cells in PBMCs. IFN-gamma stimulation 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 the milk allergy group, Br1 counts were not different between the control and the casein stimulation but significantly increased in the IFN-gamma + casein stimulated cells, and the Br1 fractions were decreased after casein stimulation and recovered in the addition of IFN-gamma for stimulation. In the milk-tolerant group, Br1 counts increased in the casein stimulated cells and in the IFN-gamma + casein stimulated cells, but the increase was significantly less when IFN-gamma was added, and the Br1 fractions were increased after casein stimulation and IFN-gamma + casein stimulation, that was not significant when IFN-gamma was added. CONCLUSIONS: IFN-gamma-induced allergen-specific Br1 responses in the PBMCs of milk allergy patients play a role in milk allergen-specific tolerance induction in vitro. Further investigations into the molecular immunological mechanisms underlying the induction of allergen-specific Br1 responses are needed.