TUBB1+ cytotoxic lymphocyte states track disease course in thyroid-associated ophthalmopathy

Background Thyroid-associated ophthalmopathy (TAO) is an orbital autoimmune disease related to Graves' disease, pathologically characterized by immune-mediated fibroinflammatory changes, which can lead to facial disfigurement and vision loss. But peripheral immune states linked to tissue remodeling remain unclear, limiting the development of effective therapeutic strategies. Methods This study enrolled a total of 29 healthy volunteers and 27 TAO patients, from whom peripheral blood was collected and peripheral blood mononuclear cells (PBMCs) were isolated. We profiled PBMCs from 6 TAO patients and 3 healthy controls by single-cell RNA sequencing and performed high-resolution subclustering of NKT and NK compartments. Furthermore, the aforementioned analytical findings were systematically validated clinically and their diagnostic efficacy was evaluated using flow cytometry. Results TAO showed selective expansion of a TUBB1⁺ NKT state enriched for integrin–cytoskeletal and chaperone-mediated autophagy programs. In parallel, a TUBB1⁺ NK subset was increased and exhibited transcriptional enrichment of wound healing, coagulation, and hemostasis pathways. Analysis of two independent orbit-tissue transcriptomic datasets highlighted cell junction/adhesion remodeling, conceptually aligning with the peripheral adhesion signatures. Flow cytometry in an independent cohort confirmed altered abundance dynamics and showed that TUBB1 ^hi NKG7 ^hi NK frequency correlated with TAO course and discriminated disease status. Conclusions These findings identify a TAO-associated peripheral cytotoxic lymphocyte program marked by TUBB1 and NKG7 that links adhesion/cytoskeletal remodeling with repair-related inflammatory pathways and may inform blood-based monitoring of TAO progression.