Effect of miR-30e regulating NK cell activities on immune tolerance of maternal-fetal interface by targeting PRF1.

AIM: Natural killer (NK) cells, as key regulatory cells, accumulate at the maternal-fetal interface in large numbers. This study explored the effect of miR-30e on regulating the activity and function of peripheral blood NK cells (PB-NK cells) and decidua NK cells (D-NK cells) by targeting PRF1 in immune tolerance of maternal-fetal interface.

METHODS: Expressions of miR-30e in PB and decidua tissues from 49 patients with recurrent spontaneous abortion and 52 normal pregnant women were measured using PCR. NK cells were isolated from PB and decidua tissues and identified by flow cytometry (FCM). In PB-NK cells and D-NK cells activated by IFN-α, expressions of miR-30e and PRF1 were determined by PCR and Western blot. Negative controls of miR-30e mimics/inhibitors and siRNA against PRF1 were transfected in PB-NK cells and D-NK cells. Expressions of miR-30e and PRF1 were determined and their relationship was verified. Expressions of KIR2DL1, NKp44, IFN-γ, TNF-α, IL-4 and IL-10 were determined by FCM. Cytotoxicity kit was used to identify the cytotoxicity of NK cells. PCR and ELISA were employed to measure expression of VEGF, Ang-2 and PGF in D-NK cells.

RESULTS: After activation by IFN-α, D-NK cells and PB-NK cells showed decreased miR-30e expression and increased PRF1 expression in normal non-pregnant women. PRF1 is a target gene of miR-30e and miR-30e negatively regulated PRF1 expression. The treatment of miR-30e mimics elevated KIR2DL1 expression and decreased NKp44 expression in PB-NK or D-NK cells. Moreover, up-regulation of miR-30e expression suppressed cytotoxicity, corresponding to increased expression of IL-4and IL-10 and reduced expression of IFN-γ and TNF-α in PB-NK and D-NK cells, as well as enhanced expression of VEGF, Ang-2 and PGF in D-NK cells. Transfection of miR-30e inhibitors could reverse the tendencies.

CONCLUSION: Up-regulated miR-30e can reduce the cytotoxicity of PB-NK cells and D-NK cells by targeting PRF1, whereby inhibiting Th1 tolerance phenotype and inducing Th2 immunodominance. miR-30e may be contributive to creating a micro-immune tolerance environment of maternal-fetal interface.