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Enteric Toll-like receptor 7 stimulation causes acute exacerbation in lupus-susceptible mice

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Clinical Rheumatology Aims and scope Submit manuscript

Abstract

Autoimmune diseases are often accompanied by acute exacerbation. However, the mechanism underlying systemic lupus erythematosus (SLE) flares remains unclear. We investigated whether short-term enteric Toll-like receptor 7 (TLR7) stimulation can exacerbate SLE using B6SKG mice, which spontaneously develop SLE due to a mutation in the zeta‒chain‒associated protein kinase 70 (Zap70) gene. Imiquimod (IMQ) or phosphate-buffered saline (PBS) were orally administered on B6WT and B6SKG mice every other day for 2 weeks. SLE exacerbation was assessed via fluorescent immunohistochemical staining of glomeruli for IgG and C3, hematoxylin and eosin staining of kidneys, and enzyme-linked immunosorbent assay for antinuclear antibody (ANA). Flow cytometry was used to evaluate germinal center B cells (GCBs), plasma cells, follicular helper T cells (Tfhs), regulatory T cells (Tregs), effector T cells (Th1s and Th17s), plasmacytoid dendritic cells (pDCs), conventional dendritic cells (cDCs), and macrophages (Mφs) in spleens. Oral administration of IMQ every other day for 2 weeks resulted in exacerbation of splenomegaly, increased IgG and C3 deposition in glomeruli, and increased ANA production in the B6SKG IMQ (SKG-IMQ) group compared to the B6SKG PBS (SKG-PBS) group; the percentages of GCBs, plasma cells, Tfhs, Th1s, pDCs, and Mφs were also increased in the SKG-IMQ group. Splenomegaly, IgG, and C3 deposition in glomeruli, and the percentages of GCBs, plasma cells, Tfhs, and Th1s were enhanced in SKG-IMQ mice compared with B6SKG mice topically treated with IMQ (SKG-ear-IMQ). Oral TLR7 stimulation in a Zap70 genetic mutation background can cause acute exacerbations of SLE.

Key Points

• The mechanism of SLE flares is not well understood.

• We have created a model that causes short-term SLE exacerbations in mice with a genetic background.

• IMQ administered orally causes more SLE in mice than transdermally.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Ms. Atsuko Tamamoto (Osaka Metropolitan University) for her technical assistance.

Funding

This study was supported by Grants-in-Aid (grant numbers: 16K09890 and 20K08773) for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (M. Hashimoto) and Yakult Bio-Science Foundation.

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Authors and Affiliations

  1. Department of Rheumatology and Clinical Immunology, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Sakyo-Ku Kyoto, Shogoin, 606-8507, Japan

    Yudai Takase, Mirei Shirakashi, Yuri Nishida, Masao Katsushima, Ryosuke Hiwa, Hideaki Tsuji, Koji Kitagori, Shuji Akizuki, Ran Nakashima, Hajime Yoshifuji & Akio Morinobu

  2. Department of Clinical Immunology, Graduate School of Medicine, Osaka Metropolitan University, Osaka, Japan

    Masao Katsushima & Motomu Hashimoto

  3. Department of Advanced Medicine for Rheumatic Diseases, Graduate School of Medicine, Kyoto University, Kyoto, Japan

    Hideo Onizawa, Akira Onishi & Masao Tanaka

  4. Division of Clinical Immunology and Cancer Immunotherapy, Center for Cancer Immunotherapy and Immunobiology, Graduate School of Medicine, Kyoto University, Kyoto, Japan

    Kosaku Murakami

  5. Department of Drug Discovery Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Japan

    Tatsuaki Tsuruyama

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  1. Yudai Takase
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Contributions

Y.T. conducted experiments. M.S. and M.H. designed experiments and supervised the project. Y.T., M.S., and M.H. analyzed and interpreted the data and wrote the manuscript. Y.N. and M.K. assisted with the experiments. T.T. evaluated histological data. All the authors contributed to the discussions and provided critical feedback for drafting the manuscript.

Corresponding author

Correspondence to Mirei Shirakashi.

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Ethics approval

All mice were maintained under specific pathogen-free conditions in our animal facility in accordance with the guidelines for animal care approved by Kyoto University (approval number: MedKyo 21535, 22116). The manuscript does not contain clinical studies or patient data.

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Takase, Y., Shirakashi, M., Nishida, Y. et al. Enteric Toll-like receptor 7 stimulation causes acute exacerbation in lupus-susceptible mice. Clin Rheumatol 42, 1185–1194 (2023). https://doi.org/10.1007/s10067-022-06467-7

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  • DOI: https://doi.org/10.1007/s10067-022-06467-7

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