Allergen alters IL‐2/αIL‐2‐based Treg expansion but not tolerance induction in an allergen‐specific mouse model

BACKGROUND: Regulatory T lymphocytes (Treg) play an important role in preventing allergic diseases. We characterized Treg expansion kinetics, marker profiles, and recirculation behavior in allergen‐challenged mice, which had been pretreated with IL‐2/αIL‐2 complexes in the presence or absence of allergen. Moreover, the ability of induced Treg to control airway hyperreactivity and effector functions of lung T cells was determined. METHODS: Humanized TCR/HLA‐transgenic allergy mice were treated in vivo with recombinant IL‐2 complexed to the anti‐IL‐2 mAb JES6‐1 in the presence or absence of mugwort pollen extract (MPE) on days 0‐2. Afterward, they were intranasally challenged with MPE (days 13‐15) followed by determination of airway hyperreactivity and lung T cell effector functions. Multiparametric flow cytometry on peripheral blood T cells was performed on a daily basis. RESULTS: IL‐2/αIL‐2 complexes highly efficiently expanded peripheral Treg cells, while concomitant allergen exposure altered the phenotype of expanded Treg cells. Notably, application of allergen together with IL‐2/αIL‐2 complexes induced expression of Treg marker molecules CTLA4, NRP1, Helios, and GITR on conventional T cells. Apart from CD25, GARP was identified as the most reliable surface‐expressed lineage discrimination marker of Treg expanded in the presence of IL‐2/αIL‐2 complexes and allergen. Finally, IL‐2/αIL‐2 complex‐expanded Treg cells could be recalled upon allergen challenge, which was associated with suppression of lung‐specific Th2 responses long after initial treatment. CONCLUSION: The characterization of reliable surface and transcription markers of IL‐2/αIL‐2 complex‐expanded Treg along with their expansion kinetics and function will help to identify protocols for their long‐term expansion in vivo.