Evaluation of the Effects of 1,25VitD3 on Inflammatory Responses and IL-25 Expression.

Nana Li,Houshang Rafatpanah,Zahra Ghaneifar,Elham Abdollahi,Seyed Abdorahim Rezaee,Nafiseh Saghafi

doi:10.3389/fgene.2021.779494

Abstract

VitD3 may contribute to a successful pregnancy through modulation of immune responses, so VitD3 deficiency may have a role in the immunopathogenesis of unexplained recurrent spontaneous abortion (URSA). However, the mechanisms of immunomodulatory actions of VitD3 in decreasing the risk of recurrent spontaneous abortion have not been understood well. Objective: The purpose of this research was to investigate the influence of 1,25VitD3 on IL-25 and related cytokines of Th17 cells including IL-17A, IL-6, and IL-23 in peripheral blood mononuclear cells of healthy women as a control group and women with unexplained recurrent spontaneous abortion. Method: Isolation of peripheral blood mononuclear cells (PBMCs) was performed from peripheral blood of the subjects of the studied groups (20 women with URSA as a case group, and 20 control women). The effects of 1,25VitD3 (50 nM, for 24 h) on the studied parameters were evaluated and were compared to the positive and negative controls in vitro. Flow cytometry analysis was used to determine the percentages of regulatory T cells and Th17 cells. For gene expression measurement and cytokines assay, real-time PCR and ELISA were carried out. Results: The proportion of Th17 cells in women with URSA was considerably higher than in the control group. IL-25 mRNA and protein levels in cultured PBMCs from women with URSA were lower than the controls. 1,25VitD3 increased IL-25 expressions at both the protein and mRNA levels in PBMCs from women with URSA relative to the control group. Additionally, 1,25VitD3 treatment not only significantly decreased the percentage of Th17 cells frequency but also reduced expressions of IL-6, IL-17A, and IL-23 in PBMCs from women with URSA. Conclusion: 1,25VitD3 may diminish inflammatory responses cells via downregulation of IL-25 expression. It could be an interesting subject for future researches in the field of the immunopathology of URSA to identify molecular pathways in URSA treatment.

Highlights

One of the most serious pregnancy complications is unexplained recurrent spontaneous abortion (URSA), which refers to three or more consecutive pregnancy losses before 20 completed gestation weeks (Dimitriadis et al, 2020)
We evaluated the frequency of Th17 cells, as well as the levels of IL-25, IL-17A, and IL-6 as related cytokines in Peripheral blood mononuclear cells (PBMCs) from women with URSA and controls, as well as the possible correlations
We found that IL-25 levels in serum and supernatants of cell culture were considerably lower in women with URPL than in the healthy women, while IL-17A, IL-23, and IL-6 levels were significantly higher compared to the controls

Summary

Introduction

One of the most serious pregnancy complications is unexplained recurrent spontaneous abortion (URSA), which refers to three or more consecutive pregnancy losses before 20 completed gestation weeks (Dimitriadis et al, 2020). Interleukin-25 (IL-17E) is a cytokine in the IL-17 family with a sequence that is 16–20% identical to that of IL-17A (Pan et al, 2001; Kolls and Lindén, 2004; Iwakura et al, 2011). In terms of structure and biological function, IL-25 is distinct from other members of the IL-17 family (Lee et al, 2001; Pan et al, 2001; Kolls and Lindén, 2004; Iwakura et al, 2011; Mantani et al, 2015). CD4+T cells, which include T helper 1 (Th1), Th2, regulatory T cells (Tregs), and Th17 cells, play an important role in the maternal immune response (Caruso et al, 2009; Saito et al, 2010; Figueiredo and Schumacher, 2016; Barad et al, 2017; Cyprian et al, 2019; Pei et al, 2019; Muyayalo et al, 2020)

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Discussion

Conclusion