CP-25 alleviates antigen-induced experimental Sjögren’s syndrome in mice by inhibiting JAK1-STAT1/2-CXCL13 signaling and interfering with B-cell migration

Abstract

The etiology of primary Sjögren’s syndrome (pSS) remains unknown, and there is no complete curative drug. In this study, we treated a mouse model of the submandibular gland (SG) protein-immunized experimental Sjögren’s syndrome (ESS) with paeoniflorin-6′-O-benzene sulfonate (termed CP-25) to evaluate the potential therapeutic effects of CP-25. Through in vivo experiments, we found that CP-25 increased saliva flow, alleviated the salivary gland indexes, and improved tissue integrity in the ESS model. The viability of splenocytes and B-lymphocyte migration from spleen were reduced in ESS mice. Furthermore, CP-25 decreased the expression of IgG antibodies, anti-SSA and anti-SSB antibodies and modulated the levels of cytokines in the serum of SS mice. The numbers of total B lymphocytes, plasma cells (PCs), and memory B cells diminished in the salivary gland. Increased expression of the JAK1-STAT1-CXCL13 axis and IFNα was found in human tissue isolated from pSS patients. In vitro, after stimulation with IFNα, the levels of CXCL13 mRNA and CXCL13 in human salivary gland epithelial cells (HSGEC) increased, while CP-25 counteracted the secretion of CXCL13 and downregulated the expression of CXCL13. IFN-α activated the JAK1-STAT1/2-CXCL13 signaling pathway in HSGEC, which was negatively regulated by additional CP-25. As a consequence, B-cell migration was downregulated in coculture with IFN-α-stimulated HSGEC. Taken together, this study demonstrated that the therapeutic effects of CP-25 were associated with the inhibition of the JAK1-STAT1/2-CXCL13 signaling pathway in HSGEC, which impedes the migration of B cells into the salivary gland. We identified the underlying mechanisms of the therapeutic effect of CP-25 and provided an experimental foundation for CP-25 as a potential drug in the treatment of the human autoimmune disorder pSS.

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Fig. 1: The ESS model was built using a modified method with submandibular gland (SG) protein immunization on day 0, 7, 14.
Fig. 2: The effects of CP-25 on saliva flow, the salivary gland indexes, and histological assessment in SS mice.
Fig. 3: The effects of CP-25 on splenocyte viability and B-lymphocyte subset from spleen in SS mice.
Fig. 4: The effects of CP-25 on the autoantibodies and cytokines from serum in SS mice.
Fig. 5: The effects of CP-25 on total B lymphocytes, memory B cells, and PCs in the salivary gland in SS mice.
Fig. 6: The expression of CXCL13 in mice serum and salivary gland.
Fig. 7: The effects of chemokines CXCL13 in SGEC stimulated by IFNa, and the expression of CXCL13 and IFNa in the human labial gland.
Fig. 8: The effect of CP-25 on the JAK1-STAT1/2-CXCL13 signaling pathway in SGECs stimulated by IFNa.

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Funding

This study was supported by the National Nature Science Foundation of China (Nos 81673444 and 81302784), Provincial Natural Science Research Project of Anhui Province (Nos KJ2017A182 and KJ2018A0166), and Key Projects of Anhui Province University Outstanding Youth Talent Fund (gxyqZD2018023).

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WW designed the study and acted as a research coordinator. HK edited the paper. HW designed the study, conducted the experiments, collected and analyzed the data, and wrote the paper. XC and FG conducted the experiments and collected and analyzed the data. SX, PZ, QL, QZ, XW, and CW helped to conduct the experiments. The final paper has been read and approved by all authors.

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Correspondence to Huaxun Wu or Wei Wei.

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Wu, H., Chen, X., Gu, F. et al. CP-25 alleviates antigen-induced experimental Sjögren’s syndrome in mice by inhibiting JAK1-STAT1/2-CXCL13 signaling and interfering with B-cell migration. Lab Invest (2020). https://doi.org/10.1038/s41374-020-0453-0

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