hPMSCs inhibit the expression of PD-1 in CD4+IL-10+ T cells and mitigate liver damage in a GVHD mouse model by regulating the crosstalk between Nrf2 and NF-κB signaling pathway

Abstract

Background The activation of T cells and imbalanced redox metabolism enhances the development of graft-versus-host disease (GVHD). Human placenta-derived mesenchymal stromal cells (hPMSCs) can improve GVHD through regulating T cell responses. However, whether hPMSCs balance the redox metabolism of CD4+IL-10+ T cells and liver tissue and alleviate GVHD remains unclear. This study aimed to investigate the effect of hPMSC-mediated treatment of GVHD associated with CD4+IL-10+ T cell generation via control of redox metabolism and PD-1 expression and whether the Nrf2 and NF-κB signaling pathways were both involved in the process. Methods A GVHD mouse model was induced using 6–8-week-old C57BL/6 and Balb/c mice, which were treated with hPMSCs. In order to observe whether hPMSCs affect the generation of CD4+IL-10+ T cells via control of redox metabolism and PD-1 expression, a CD4+IL-10+ T cell culture system was induced using human naive CD4+ T cells. The percentage of CD4+IL-10+ T cells and their PD-1 expression levels were determined in vivo and in vitro using flow cytometry, and Nrf2, HO-1, NQO1, GCLC, GCLM, and NF-κB levels were determined by western blotting, qRT-PCR, and immunofluorescence, respectively. Hematoxylin-eosin, Masson’s trichrome, and periodic acid-Schiff staining methods were employed to analyze the changes in hepatic tissue. Results A decreased activity of superoxide dismutase (SOD) and a proportion of CD4+IL-10+ T cells with increased PD-1 expression were observed in GVHD patients and the mouse model. Treatment with hPMSCs increased SOD activity and GCL and GSH levels in the GVHD mouse model. The percentage of CD4+IL-10+ T cells with decreased PD-1 expression, as well as Nrf2, HO-1, NQO1, GCLC, and GCLM levels, both in the GVHD mouse model and in the process of CD4+IL-10+ T cell generation, were also increased, but NF-κB phosphorylation and nuclear translocation were inhibited after treatment with hPMSCs, which was accompanied by improvement of hepatic histopathological changes. Conclusions The findings suggested that hPMSC-mediated redox metabolism balance and decreased PD-1 expression in CD4+IL-10+ T cells were achieved by controlling the crosstalk between Nrf2 and NF-κB, which further provided evidence for the application of hPMSC-mediated treatment of GVHD.

TLR4 Modulates Senescence and Paracrine Action in Placental Mesenchymal Stem Cells via Inhibiting Hedgehog Signaling Pathway in Preeclampsia

“…PMSCs adhered to cell culture plates after 3 days of culture, and the third passage of PMSCs showed fibroblastic morphology (Figure S1(a) ). The expression of surface markers CD44, CD73, CD90, and CD105 was positive, while the expression of surface markers CD31, CD34, and CD45 was negative (Figure S1(b) ), which was consistent with previous reports [ 6 , 51 , 52 ]. PMSCs also successfully formed calcified bone nodules, lipid droplets, and cartilage in the osteogenic, adipogenic, and chondrogenic differentiation assays (Figure S1(c) ).…”

“…PMSCs were incubated with PBS or fluorescein-labeled antibodies for 30 min. The tested antibodies included CD90, CD 105, CD73, CD44, CD34, CD31, and CD45 (Biolegend, USA) as previously reported [ 6 , 51 , 52 ]. Isotype-identical antibodies were used as controls.…”

PFKFB3-mediated glycometabolism reprogramming modulates endothelial differentiation and angiogenic capacity of placenta-derived mesenchymal stem cells

“…Written informed consents were also obtained. Based on previously described methods [ 19 , 27 , 28 ], PMSCs were isolated from term placentas collected from healthy donors who had gone through elective cesarean sections due to the breech position or previous history of cesarean section. The placental tissue was taken from the center area around the umbilical cord attachment site and was gently washed with preheated phosphate-buffered saline (PBS, pH 7.2 ± 0.1, Genome Sciences), minced into 1mm 3 pieces, and digested with gentle shaking in 0.1 mg/mL type I collagenase (Gibco) for 60 min at 37 °C.…”

“…Cell suspensions were incubated with PBS or fluorescein-labeled antibodies for 30 min. The tested antibodies included CD73, CD90, CD105, CD31, CD34, CD45, and HLA-DR (BD Biosciences, USA) as previously reported [ 19 , 27 , 28 ]. Isotype-identical antibodies were used as controls.…”

Mesenchymal stem cell homing to improve therapeutic efficacy in liver disease

“…Yoo et al showed that MSCs suppress T cells by inhibiting CD25 translation through the LKB1-AMPK-mTOR pathway [ 14 ]. Furthermore, Zhang et al found that human placenta (hP)-MSCs could inhibit PD-1 expression in CD4 + IL-10 + T cells and alleviate liver injury in a graft versus host disease mouse model by regulating the crosstalk between Nrf2 and NF-κB signaling pathways [ 15 ]. Regulatory T cells (Tregs) play a crucial role in inhibiting immune cell-mediated hepatocyte injury during fulminant hepatitis [ 40 ].…”