Phenotype analyses of IL-10-producing Foxp3- CD4+ T cells increased by subcutaneous immunotherapy in allergic airway inflammation

Abstract

Introduction: The mechanisms of allergen immunotherapy are not fully elucidated. Here, we sought to develop a murine model to demonstrate the effectiveness of subcutaneous immunotherapy (SCIT) for allergic responses. As excessive antigen dosages may induce immune tolerance in sensitized mice, the effects of SCIT were assessed by varying the antigen dosage. The mechanisms of SCIT were analyzed by focusing on the induction of Foxp3⁺ Treg cells and IL-10-producing Foxp3^- CD4⁺ T cells, as well as on the phenotype of the latter cells.

Methods: Ovalbumin (OVA) + Al(OH)₃-sensitized mice received subcutaneous dosages of OVA at 0.01, 0.1 or 1 mg/animal for SCIT, followed by intratracheal challenges with OVA at 5, 50 or 500 μg/animal.

Results: The maximum effects of SCIT were observed with 1 mg/animal of OVA for airway inflammation induced by 5 μg/animal of OVA, in which airway eosinophilia and Th2 cytokine production were markedly suppressed. The increase in the OVA-specific IgE level was significantly suppressed by SCIT. The development of bronchial epithelial thickening and mucus accumulation were also suppressed by SCIT. Concomitantly, IL-10-producing Foxp3^- CD4⁺ T cells were increased in the lungs by SCIT, but Foxp3⁺ Treg cells were not. Most of the induced IL-10-producing Foxp3^- CD4⁺ T cells were negative for either IL-5 or LAG-3, but positive for CD49b.

Conclusion: We successfully developed an airway allergic model for SCIT. It was suggested that most of IL-10-producing Foxp3^- CD4⁺ regulatory T cells increased by SCIT in the lungs were CD49b⁺ CD4⁺ regulatory T cells, but neither Th2 cells nor Tr1 cells.

Keywords: Airway inflammation; Allergen immunotherapy; Interleukin-10; Regulatory T cell; Th2 cell; Type 1 regulatory T cell.