Glucose-Responsive CD20+ Cytotoxic T Cells: A Novel Pro-inflammatory Mediator in the Immunopathogenesis of Type 2 Diabetes

Albayrak, O.; Batman, A.; Unlu, S.; Akkaya, N.; Khan, S. S.; Demir, S. C.; Ozisik, S.; Sezer, H.; Yilmaz, Z. Y.; Mizrak, B.; Degneli, O.; Yazici, D.; Vural, A.

doi:10.64898/2026.02.12.705577

bioRxiv · 10.64898/2026.02.12.705577

Glucose-Responsive CD20+ Cytotoxic T Cells: A Novel Pro-inflammatory Mediator in the Immunopathogenesis of Type 2 Diabetes

Albayrak, O.·Batman, A.·Unlu, S.·Akkaya, N.·Khan, S. S.·Demir, S. C.·Ozisik, S.·Sezer, H.·Yilmaz, Z. Y.·Mizrak, B.·Degneli, O.·Yazici, D.·Vural, A.

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Abstract

ObjectiveAlthough T-cell-mediated inflammation is a hallmark of Type 2 Diabetes (T2D), the contribution of CD20-expressing T cells--a highly potent subset recently implicated in various autoimmune conditions--to T2D pathogenesis remains unknown. This study aimed to characterize the frequency, functional profile, and glucose-responsiveness of CD20+ cytotoxic T lymphocytes (CTLs) in patients with T2D. Research Design and MethodsPeripheral blood mononuclear cells (PBMCs) from 25 treatment-naive T2D patients and 20 age-matched healthy controls (HC) were analyzed using multicolor flow cytometry. We assessed the frequency of CD3+CD8+CD20+ cells and their production of cytotoxic molecules (Granzyme B, Perforin, Granzyme K) and pro-inflammatory cytokines (IFN-{gamma}, TNF-, GM-CSF). To further establish the link with hyperglycemia, HC-derived CTLs were exposed to increasing glucose concentrations (100-450 mg/dL) in vitro. Single-cell RNA sequencing (scRNA-seq) data from a public T2D dataset was utilized to validate the molecular signature of MS4A1 (CD20)+ CTLs. ResultsThe frequency of circulating CD20+ CTLs was significantly elevated in T2D patients compared to HCs (p<0.0001) and demonstrated a strong positive correlation with HbA1c, fasting glucose, and triglyceride levels. Notably, CD20+ CTLs from T2D patients exhibited a "hyperfunctional" phenotype, characterized by significantly higher degranulation (CD107a), elevated expression of Granzymes B/K and Perforin, and increased production of IFN-{gamma} and TNF- compared to HCs (p<0.01 for all). In contrast, no such differences were observed in the CD20-CTL compartment. In vitro experiments revealed that escalating glucose levels directly enhanced the proliferation and cytotoxic potential of CD20+ CTLs, suggesting a nutrient-sensing mechanism. scRNA-seq analysis further confirmed the distinct pro-inflammatory and effector transcriptional profile of MS4A1+ T cells in T2D. ConclusionsOur findings identify CD20+ CTLs as a novel, glucose-sensitive, and hyperfunctional immune subset in T2D. The strong correlation between these cells and clinical metabolic parameters suggests that CD20+ CTLs may act as a critical link between chronic hyperglycemia and systemic inflammation, representing a potential new therapeutic target for immunomodulation in T2D.

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Albayrak, O., Batman, A., Unlu, S., Akkaya, N., Khan, S. S., Demir, S. C., Ozisik, S., Sezer, H., Yilmaz, Z. Y., Mizrak, B., Degneli, O., Yazici, D., Vural, A.. 2026-02-13. Glucose-Responsive CD20+ Cytotoxic T Cells: A Novel Pro-inflammatory Mediator in the Immunopathogenesis of Type 2 Diabetes. https://doi.org/10.64898/2026.02.12.705577

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