Immunopharmacological properties of VitD3: 1,25VitD3 modulates regulatory T cells and Th17 cells and the cytokine balance in PBMCs from women with unexplained recurrent spontaneous abortion [URSA]

Abstract

Background: 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 regulatory T cells /Th17 axis, the gene expressions and concentrations of related cytokines including, TGF-β, IL-10, IL-6, IL-23, and IL-17A in peripheral blood mononuclear cells [PBMCs] of healthy women as a control group and women with URSA. Method: Isolation of 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 hours] 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 regulatory T cells was markedly lower, while the proportion of Th17 cells in women with URSA was considerably higher than in the control group [P=0.01, P=0.01]. The ratio of the frequency of Tregs to the baseline [1,25VitD3/Untreated] increased, while the ratio of the frequency of Th17 cells to the baseline decreased in women with URSA relative to the controls [P= 0.01, P=0.04]. 1,25VitD3 increased IL-10 expressions at both the protein and mRNA levels in PBMCs in women with URSA relative to the control group [P=0.0001, P=0.04]. TGF-β levels in the cultured supernatants decreased significantly in the case group in the presence of 1,25VitD3 relative to the controls [P=0.03]. 1,25VitD3 treatment also significantly decreased gene expressions of IL-6, IL-17A, and IL-23 in PBMCs of women with URSA [P=0.01, P=0.001, P=0.0005], as well as the levels of those cytokines in cell culture supernatants [P=0.03, P=0.02, P=0.01, respectively] in women with URSA relative to the controls. Conclusion: According to the findings of this research, modulation of immune responses by 1,25VitD3 is accomplished by strengthening Tregs function and inhibiting inflammatory responses of Th17 cells which may have a positive impact on pregnancy outcome. Thus, as an immunomodulating agent, VitD3 may be effective in reducing the risk of URSA.