Background:Early administration of IL-6R blockade is effective in adult-onset Still's disease (AOSD), but the underlying immune alterations during combined immunotherapy remain unclear.
Methods:We employed high-dimensional single-cell mass cytometry and an unbiased bioinformatics pipeline to characterize the immune landscape in treatment-naïve AOSD patients, stable AOSD patients after 24 weeks of IL-6R blockade plus methotrexate and prednisone, and matched healthy controls. Cytokine profiling was conducted using a high-throughput cytometric bead array, and potential regulatory networks were identified using the PerformanceAnalytics package. Validation was performed via flow cytometry and mRNA sequencing.
Results:We identified 22 peripheral immune cell populations and characterized their composition and phenotypic markers. Treatment-naïve AOSD patients exhibited significant depletion of CD4+ T cells and B cells, along with excessive activation of CD8+ T cells and a previously unreported CD45+CD3-CD19-CD10-CD66a+ population, findings that were validated in an independent AOSD cohort. mRNA sequencing revealed the proinflammatory role of CD8+ T cells. Notably, these dysregulated immune profiles were markedly restored following immunotherapy. IL-18, IL-21, and IFN-γ demonstrated strong associations with adaptive immune cells and AOSD clinical indices.
Conclusions:Combined IL-6R blockade effectively modulates immune dysregulation in refractory AOSD, reversing key pathological immune alterations and highlighting its therapeutic potential.
Keywords:Adult-onset Still’s disease; Mass cytometry; Tocilizumab.

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