Bone marrow mesenchymal stromal cells attenuate liver allograft rejection may via upregulation PD-L1 expression through downregulation of miR-17-5p

Abstract

Studies have reported that bone marrow mesenchymal stem cells (BMSCs) play an important role in immune regulation after organ transplantation. Some of the BMSC-mediated regulatory mechanisms are related to PD-L1 expression. However, the regulatory mechanism of PD-L1 expression is not fully understood. In this experiment, we confirmed the regulatory role of BMSCs in the rejection of liver transplantation and explored the possible mechanism by which PD-L1 expression is regulated in BMSCs. An orthotopic liver transplantation model in rats was established based on the "double-cuff technique". Third-generation BMSCs were injected into the rejection model rats via portal vein. At the same time, sera were obtained from rats in the allograft, isograft and sham-operated groups. The sera from these groups were separately added to BMSCs complete medium to partially mimic the in vivo environment in which BMSCs are exposed. After predicting and analysing microRNAs that regulate PD-L1 expression, we examined the microRNA and PD-L1 expression in BMSCs of the three groups cultured in the conditioned media. Moreover, a luciferase reporter assay was used to confirm the interaction between PD-L1 and microRNA. The study found that the liver function and liver graft histopathology of the BMSC-treated group were better than those of the allograft group. The Kaplan-Meier survival curve analysis showed that the median survival time (MST) of the BMSC-treated group was longer than that of the allograft group. Bioinformatics prediction and analysis showed that miR-17-5p is highly likely to regulate PD-L1. Compared with the other two groups of cells, BMSCs cultured with serum from allograft models showed higher PD-L1 expression, but lower miR-17-5p expression. Pearson correlation analysis showed that miR-17-5p and PD-L1 expression was negatively correlated, and further luciferase reporter assays confirmed that miR-17-5p interacted directly with the 3´-untranslated region (UTR) of PD-L1 mRNA. The results of this study demonstrate that BMSCs can attenuate liver allograft rejection and provide us with the idea that BMSCs may further upregulate PD-L1 expression by downregulating miR-17-5p expression, thereby attenuating liver allograft rejection.

Keywords: Bone marrow mesenchymal stem cells (BMSCs); Liver transplantation; PD-L1/B7-H1/CD274; Rejection; miR-17-5p.