RESUMO
Objective To analyse the change of DNA methylation with 5-Azac injection in acute graft-versus-host dis- ease (aGVHD) mouse model, which received allogeneic bone marrow transplantation, and explore the immunomodulatory ef-fects of 5-Azac. Methods Male C57BL/6 (H-2b)and female BALB/c (H-2d) mice were selected as donor and recipient of complete allotransplantation. BABL/c mice were divided into two groups, transplantation control group and 5-Azac experi-mental group. At 1-7, 14, 21 and 28-day after transplantion, 5-Azac 0.25 mg/kg (0.3 mL/time) was injected by tail vein in experimental group, while the control group were injected with sterile water 0.3 mL/time. Peripheral blood DNA samples were collected from three control mice and three experimental mice, then mixed with equal amount respectively. The MeDIP-seq method was selected to detect methylation changes in mice, and the differential DNA methylation in the biological path-ways was analyzed. Results The survival time was prolonged, and the rejection reaction was decreased in 5-Azac experi-mental group, which suggested immune hyporesponsiveness post aGVHD. The MeDIP-seq result showed that 369 different DNA methylation located in the promoter regions, including 239 up-regulated genes and 130 down-regulated genes. There were 184 differential DNA methylation genes located in the exon regions, including 113 up-regulated genes and 71 down-regulated genes. Differential DNA methylation genes involved in 10 immunological signaling pathways, respectively. Among them, TGF-β, GSK-3β, SYK, PI3K, NFAT, CD28 andα4β7 were closely related to the development of aGVHD. Conclu-sion 5-Azac can effectively induce immune hyporesponsiveness post aGVHD by changing the gene methylation status.
RESUMO
Objective To study the expression of immunoglobulin-like transcripts 3 (ⅡT3) and ILT4 in peripheral blood dendritic cells (DC) of kidney transplantation recipients and to analyze its significance in immunity hyporesponsiveness of transplantation. Methods Twenty kidney allograft recipients who were survived more than five years were recruited to two groups: renal function stable groups, chronic rejection groups, and 10 healthy volunteers served as a control group. The peripheral blood mononuclear cells (PBMC) were stimulated with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) and immature DC were obtained. The expression of ILT3 and ILT4 was detected by using flow cytometry. The level of HLA-G5 in serum was determined by using enzyme linked immunosorbent assay. Results ILT3 expression in renal function stable group was increased and decreased in chronic rejection groups as compared with control group (P＜0.05),but ILT4 expression had no significant difference among all groups. HLA-G5 in serum was significantly increased in renal function stable group as compared with other groups. Conclusion Expression of ILT3 and HLA-G was increased in the kidney transplantation recipients with stable renal function and long-term survival, suggesting that they may play an important role in inducing and maintaining periphery immune tolerance.
RESUMO
Objective To study the mechanism of immune hyporesponsiveness of allograft rejection induced by transfusion nonpufsed allopeptide syngeneic immature dendritic cell(imDC) generated from recipient bone marrow progenitors and to explore a possible strategy for liver allograft protection in clinic.Methods Forty experimental rats were randomly divided into 4 group: control group,cyclosporine A(CsA) group,mature DC(mDC) group and imDC group.In control group,Wistar rats only received liver transplantation.In CsA group,Wistar rats underwent liver transplantation plus CsA treatment(10 mg/(kg?d)).In mDC group,recipient-derived mDC 1?106 were infused intravenously through the penile vein to Wistar rats.In imDC group,ImDC with the dose of 1?106 were injected into Wistar rats via the dorsum vein of penile.In each group,five recipients were killed on the 10th day after transplantation,the other five recipients were left to observe survival time.The levels of ALT,AST,TBIL,IL-2,IFN-?,IL-4 and IL-10 were detected.The acute rejection and the expression of FasL/Fas in the grafts were detected by HE and immunohistochemical staining.Western blot was used to detect Scurfin protein expression of CD4+ CD25+ T cells.Results The median survival time of the liver allografts in CsA group and imDC group were significantly longer than that in control group and mDC group(P