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Calycosin synergizes with methotrexate in the treatment of Sjögren’s disease by targeting BATF in T follicular helper cells

Acta Pharmacologica Sinica volume 46, pages 1990–2005 (2025)Cite this article

Abstract

T follicular helper (Tfh) cells are crucially involved in the pathogenesis of autoimmune disorders, including Sjögren’s disease (SjD, also known as Sjögren’s syndrome), by promoting effector B cell responses and autoantibodies production. However, targeting Tfh cells remains challenging. In this study, we identified that calycosin (Caly), a natural flavonoid, effectively suppressed pathogenic Tfh cell responses, although it did not affect the plasmacytic differentiation of B cells. Under Tfh polarization conditions, Caly rapidly bound to the master transcription factor, BATF, in both human and murine CD4+ T cells and thus potently disrupted BATF-mediated Maf gene transcription. Methotrexate (MTX), a first-line medication in the treatment of autoimmune disorders, mainly suppresses B cell responses but fails to target Tfh cells. In a mouse model of experimental Sjögren’s syndrome (ESS) that we previously established, MTX synergized with Caly in attenuating the disease pathology and autoantibodies in ESS mice with chronic inflammation, with signs of disease remission. This immunomodulatory function was also validated in peripheral blood mononuclear cells from patients with SjD. Thus, Caly may serve as a novel inhibitor of BATF in suppressing Tfh-cell-mediated humoral autoimmunity and elicit a synergistic effect in combination with B-cell-targeting strategies.

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Fig. 1: Caly treatment alleviated the development of ESS in mice.
Fig. 2: Caly inhibited the Tfh-cell-mediated humoral immune response in mice with ESS.
Fig. 3: Caly bound to BATF and disrupted c-Maf transcription upon IL-6 stimulation.
Fig. 4: Caly bound to BATF and disrupted c-Maf transcription in Tfh cells.
Fig. 5: Caly inhibited the BATF/c-Maf axis in human Tfh cells.
Fig. 6: Methotrexate synergized with Caly in the treatment of chronic ESS.
Fig. 7: MTX synergized with Caly in the suppression of effector subsets in PBMCs from patients with SjD.
Fig. 8: Schematic diagram illustration of the effect and mechanism of Caly on Tfh cells.

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Acknowledgements

We thanked the staff of Faculty Core Facility, Li Ka Shing Faculty of Medicine, the University of Hong Kong for their kind support. We thank the kind help of Dr. Sarah Pringle, University Medical Center Groningen, Netherland. This work was supported by grants through National Key Research and Development Program of China (2023YFE0203100), Mainland-Hong Kong Joint Funding Scheme (MHP/104/22), General Research Fund, Hong Kong Research Grants Council (17116521, 27111820 and 17109123), Health and Medical Research Fund (19201121 and 20212601), Hong Kong Research Grants Council Area of Excellence Scheme 2016/2017 (No.467 AoE/P-705/16).

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  1. These authors contributed equally: Su-lan Yu, Mei-ling Wu

Authors and Affiliations

  1. School of Chinese Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, 999077, China

    Su-lan Yu, Mei-ling Wu, Ya-cun Chen, Jing Xie, Xiao-yu Xu, Dan-bao Ma, Jian-gang Shen & Xiang Lin

  2. State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, Hong Kong SAR, 999077, China

    Mei-ling Wu, Jian-gang Shen & Xiang Lin

  3. Division of Rheumatology and Clinical Immunology, Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong SAR, 999077, China

    Philip Hei Li

  4. Department of Ophthalmology, Peking University First Hospital, Beijing, 100034, China

    Yun Feng

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Contributions

XL: Conceptualization, Methodology, Visualization, Writing—original draft, Writing—review & editing, Funding acquisition, Project administration, Investigation, Software, Supervision. JGS: Writing—review & editing, Funding acquisition, Supervision. SLY, MLW: Investigation, Formal analysis, Software, Validation, Writing—original draft. PHL: Resources, YCC, JX, XYX, DBM: Investigation, Validation. All authors approved the final manuscript. YF: Writing—review & editing, Funding acquisition.

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Correspondence to Jian-gang Shen or Xiang Lin.

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Yu, Sl., Wu, Ml., Li, P.H. et al. Calycosin synergizes with methotrexate in the treatment of Sjögren’s disease by targeting BATF in T follicular helper cells. Acta Pharmacol Sin 46, 1990–2005 (2025). https://doi.org/10.1038/s41401-025-01536-4

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