Single-cell Analysis of TFH Cell Subsets in Allergic Rhinitis: Discovery of the GZMK+ TFH Cell Subset

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Background: Allergic rhinitis (AR) is a prevalent condition linked to IgE-mediated immune responses. T follicular helper (TFH) cells, particularly the TFH2 subset, have been implicated in the pathogenesis of AR due to their role in promoting IgE production. However, the number, functional gene expression, and differences among the three TFH subsets in AR patients remain unclear. Methods: This study recruited six AR patients and three healthy controls. Flow cytometry and single-cell sequencing were used to identify and analyze subsets of TFH cells. TFH1, TFH2, and TFH17 subsets were identified based on CXCR3 and CXCR6 expression. Functional gene expression in TFH subsets of AR patients and healthy controls was analyzed to explore differences in immune responses. Results: AR patients exhibited a significant increase in circulating TFH2 cells compared to healthy controls, correlating with disease severity. Additionally, precursor memory TFH cells were higher in AR patients. However, no significant differences were found in the expression of functional membrane molecules on TFH cells between the two groups.. Single-cell RNA sequencing revealed nine TFH clusters with differential expression of functional genes, including a GZMK + TFH subset. The increased proportion of TFH1 and TFH2 subsets in AR compared to controls suggests their significant involvement in the pathogenesis of AR. Conclusions: Our study reveals increased TFH2 cells and differential TFH subset distributions in AR patients, especially the discovery of the GZMK + TFH subset, providing insights into TFH-mediated mechanisms in AR. These findings suggest potential therapeutic targets for AR treatment.

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