Altered chromatin landscape in circulating T follicular helper and regulatory cells following grass pollen subcutaneous and sublingual immunotherapy

Background: Allergen-specific immunotherapy is a disease-modifying treatment that induces long-term T-cell tolerance.

Objective: We sought to evaluate the role of circulating CXCR5⁺PD-1⁺ T follicular helper (cT_FH) and T follicular regulatory (T_FR) cells following grass pollen subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT) and the accompanying changes in their chromatin landscape.

Methods: Phenotype and function of cT_FH cells were initially evaluated in the grass pollen-allergic (GPA) group (n = 28) and nonatopic healthy controls (NAC, n = 13) by mathematical algorithms developed to manage high-dimensional data and cell culture, respectively. cT_FH and T_FR cells were further enumerated in NAC (n = 12), GPA (n = 14), SCIT- (n = 10), and SLIT- (n = 8) treated groups. Chromatin accessibility in cT_FH and T_FR cells was assessed by assay for transposase-accessible chromatin sequencing (ATAC-seq) to investigate epigenetic mechanisms underlying the differences between NAC, GPA, SCIT, and SLIT groups.

Results: cT_FH cells were shown to be distinct from T_H2- and T_H2A-cell subsets, capable of secreting IL-4 and IL-21. Both cytokines synergistically promoted B-cell class switching to IgE and plasma cell differentiation. Grass pollen allergen induced cT_FH-cell proliferation in the GPA group but not in the NAC group (P < .05). cT_FH cells were higher in the GPA group compared with the NAC group and were lower in the SCIT and SLIT groups (P < .01). Time-dependent induction of IL-4, IL-21, and IL-6 was observed in nasal mucosa following intranasal allergen challenge in the GPA group but not in SCIT and SLIT groups. T_FR and IL-10⁺ cT_FH cells were induced in SCIT and SLIT groups (all, P < .01). ATAC-seq analyses revealed differentially accessible chromatin regions in all groups.

Conclusions: For the first time, we showed dysregulation of cT_FH cells in the GPA group compared to NAC, SCIT, and SLIT groups and induction of T_FR and IL-10⁺ cT_FH cells following SCIT and SLIT. Changes in the chromatin landscape were observed following allergen-specific immunotherapy in cT_FH and T_FR cells.