Mesenchymal stem cell transplantation alleviates experimental Sjögren's syndrome through IFN-β/IL-27 signaling axis

Genhong Yao, Jingjing Qi, Jun Liang, Bingyu Shi, Weiwei Chen, Wenchao Li, Xiaojun Tang, Dandan Wang, Liwei Lu, Wanjun Chen, Songtao Shi, Yayi Hou, Lingyun Sun, Genhong Yao, Jingjing Qi, Jun Liang, Bingyu Shi, Weiwei Chen, Wenchao Li, Xiaojun Tang, Dandan Wang, Liwei Lu, Wanjun Chen, Songtao Shi, Yayi Hou, Lingyun Sun

Abstract

Rationale: Although mesenchymal stem cell (MSC) transplantation has been proved to be an effective therapeutic approach to treat experimental Sjögren's syndrome (SS), the detailed underlying mechanisms remains unknown. IL-27 has diverse influences on the regulation of T cell differentiation and was involved in SS through modulating immune response. Here we aimed to explore whether IL-27-mediated regulation of immune cells was responsible for the beneficial effects of MSC transplantation on SS. Methods: The SS-like symptoms were evaluated in IL-27 deficient and recombinant IL-27-treated NOD mice. The MSCs were infused into NOD mice via the tail vein. The histological features of submandibular glands, saliva flow rate and serum IL-27 were examined. The effects of MSCs on the IL-27 production and Th17/Treg cell in SS patients and mice in vitro and in vivo were determined for the mechanistic study. Results: This study showed that SS patients had decreased IL-27 level and increased ratio of Th17/Treg cells. Consistently, exacerbated SS-like symptoms were observed in IL-27 deficient NOD mice, along with increased ratio of Th17/Treg cells. Importantly, MSC transplantation alleviated SS-like symptoms by elevating the level of IL-27 to restore Th17/Treg balance in NOD mice. Mechanistically, MSC-secreted interferon-β (IFN-β) promote dendritic cells to produce IL-27. Conclusions: Thus, we have revealed a previously unrecognized function of MSC-mediated IL-27 production by DCs in suppressing SS-like syndrome, which provided evidences for clinical application of MSC in patients with SS.

Keywords: IL-27; Mesenchymal stem cells; Sjögren's syndrome.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

© The author(s).

Figures

Figure 1
Figure 1
Serum IL-27 negatively correlates with disease activity in patients with Sjögren's syndrome. (A, B) IL-27 mRNA in PBMCs (A) and serum IL-27 (B) in patients with Sjögren's syndrome (SS) (n=30) compared with those of healthy controls (HC) (n=30). (C,D) Expression of IL-27 receptors, gp130 mRNA (C) and IL-27RαmRNA (D), were detected in PBMC from SS patients (n=5) and HC. (n=5) (E) Serum IL-27 was assessed according the European League Against Rheumatism (EULAR) Sjögren's syndrome Disease Activity Index (ESSDAI) scores. (F) Serum IL-27 was compared between SS patients with (n=14) and without anti-SSA antibody (n=15). (G) Correlation of serum IL-27 and IgG was analyzed. (H) Percentages of Th17 and Treg cells in SS patients (n=15) and HC (n=15) were shown. (I, J) Serum TGF-β (i) and IL-17A (j) in SS patients and HC were detected. (K, L) Correlations of IL-27 and Treg and Th17 cells were evaluated. Data were based on three independent experiments. Data are presented as mean±SEM. *, p<0.05, **, p<0.01, ***, p<0.001.
Figure 2
Figure 2
IL-27 deficiency exacerbates SS-like symptoms in NOD mice. (A) Serum IL-27 was compared between NOD (n=5) (12-week old) and control ICR mice (n=5) (12-week old). (B) Salivary flow rates were measured in wild type (WT) (n=5) (12-week old) and IL-27-/- NOD mice(n=5) (12-week old). (C) Serum IgG in WT and IL-27-/- NOD mice was compared (n=5) (12-week old). (D) Representative images of H&E staining of salivary glands in WT and IL-27-/- NOD mice. Original magnification, ×40. Scale bars, 64 μm. Yellow arrows indicate infiltrating lymphocytes. (E) Lymphocyte infiltrations in salivary glands of mice were evaluated for histological scores. (F and G) Percentages of Treg and Th17 cells in WT (n=5) (12-week old) and IL-27-/- NOD mice (n=5) (12-week old) were shown. Data were based on three independent experiments. Data are presented as mean±SEM. *,p<0.05, **, p<0.01.
Figure 3
Figure 3
Mesenchymal stem cell (MSC) transplantation attenuates SS-like diseases in NOD mice. (A) Representative images of H&E staining of salivary glands in PBS (A) and MSC treated NOD mice. Yellow arrows indicate infiltrating lymphocytes. (B) Lymphocyte infiltrations in salivary glands of mice were evaluated for histological scores. (C) Salivary flow rates in PBS (n=5) (12-week old) and MSC treatment NOD mice (n=5) (12-week old). (D)Serum IL-27 was compared between PBS (n=5) (12-week old) and MSC treatment groups (n=5) (12-week old). (E-H) Representative FACS analysis of Treg and Th17 cells of spleens (E, F) and cervical lymph nodes (G, H)(n=5). Data were based on three independent experiments. Data are presented as mean±SEM. *,p<0.05, **, p<0.01.
Figure 4
Figure 4
IL-27 modulates the proliferation and differentiation of Th17 and Treg cells in PBMCs and naïve CD4+ T cells. (A, B) Treg and Th17 cells in human PBMCs with and without IL-27 treatment. (A) Fluorescence staining of CD4, Foxp-3 and IL-17 in human PBMCs. (B) Summary of the frequencies of Treg and Th17 cells (n=5). (C, D) Flow cytometric analysis of Treg and Th17 cells, which were differentiated from mouse naïve CD4+ T cells in the presence or absence of IL-27 (n=5). Data are presented as mean±SEM. Data were based on three independent experiments. *, p<0.05.
Figure 5
Figure 5
MSCs increase serum IL-27 and restore Th17/Treg balance in SS patients. (A, B, C) Serum IL-27 (A), IL-17A (B) and TGF-β (C) were determined in SS patients before and 1 week after MSC transplantation (n= 11). (D, E, F) Fluorescence staining of CD4, Foxp-3 and IL-17 in human PBMCs (D, F). Summary of Treg and Th17 cell frequencies in SS patients with MSC transplantation (E, G) (n=10). Data are presented as mean±SEM. Data were based on three independent experiments. *, p<0.05, **, p<0.01.
Figure 6
Figure 6
MSCs promote IL-27 production of dendritic cells through secreting IFN-β. (A) IL-27 produced by monocyte-derived DCs from SS patients (n=5) and HC (n=5). (B, C) IL-27 mRNA (B) and protein (C) of monocyte-derived DCs from HC with or without MSC treatment in the presence of 100ng/mL LPS was evaluated (n=5). (D) Genes related to IL-27 expression of DCs were screened and compared in MSCs co-cultured with and without monocyte-derived DCs from HC (n=4) in the presence of 100ng/mL LPS. (E) IL-27 mRNA expressions of monocyte-derived DCs with different concentrations of IFN-β treatment in the presence of 100ng/mL LPS (n=3). (F) IL-27 mRNA expressions of monocyte-derived DCs from HC co-cultured with MSC with and without IFN-β mRNA interference in the presence of 100ng/mL LPS (n=4). αMSC means siRNA silencing of IFN-β in MSCs. Data are presented as mean±SEM. Data were based on three independent experiments. *, p<0.05, **, p<0.01, ***, p<0.001.

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