Despite the efficacy of allergen-specific immunotherapy (AIT), the role of trained immunity and tolerance in this process has not been elucidated.

Here, we have performed a comprehensive longitudinal analysis of the systemic innate immune cell repertoire during the course of AIT.

Patients with allergy received standard preseasonal subcutaneous AIT with allergoids to birch and/or grass. Healthy controls were monitored without any intervention. Flow cytometry of innate lymphoid cell (ILC), natural killer cell, monocyte cell, and dendritic cell (DC) subsets was performed at baseline, 3 months (birch season), 6 months (grass seasons), and 12 months after the therapy in patients or at similar seasonal time points in controls. Additional analyses were performed in the third-year birch and grass season.

We observed a durable decrease in group 2 ILCs and an increase of group 1 ILCs after AIT, with dynamic changes in their composition. We found that an expansion of CD127⁺CD25⁺⁺ clusters caused observed shifts in the heterogeneity of group 1 ILCs. In addition, we observed development of CD127⁺CD25⁺⁺c-Kit⁺ group 3 ILC clusters. Moreover, we found an increase in the number of intermediate monocytes in parallel with a reduction in nonclassical monocytes during the first year after AIT. Classical and intermediate monocytes presented significant heterogeneity in patients with allergy, but AIT reduced the HLA-DR⁺⁺ clusters. Finally, an increase in plasmacytoid DCs and CD141⁺ myeloid DCs was observed in individuals with allergy, whereas the number of CD1c⁺ myeloid DCs was reduced during the first year of AIT.

AIT induces changes in the composition and heterogeneity of circulating innate immune cells and brings them to the level observed in healthy individuals. Monitoring of ILCs, monocytes, and DCs during AIT might serve as a novel biomarker strategy.