Immunological tolerance induced by Sirolimus combine with modified sTNFRI-IgGFc gene in rat dendritic cells / 中华器官移植杂志

ABSTRACT

Objective To explore the morphology,cell phenotype and cell function in dendritic cells (DCs) derived from bone marrow after treatment with Sirolimus or Sirolimus combined with sTNFRI-IgGFc gene segment transfection.Method DCs were divided into 5 groups (imDCs,mDCs,Rapa-DCs,sTNFRI-DCs and Rapa-sTNFRI-DCs) according to different interventions.The expresson of MHC-Ⅱ,CD80 and CD86 was detected by flow cytometry.T cell proliferation of the mixed lymphocyte reaction was evaluated by MTT method.The levels of IL-12,IFN-γ and sTNFRI-IgGFc were determined by ELISA.Result On the day 10,the flow cytometry showed that the expression levels of MHC-Ⅱ,CD80 and CD86 on the cell surface in Sirolimus group were significantly higher than the other groups (P＜0.05).The expression level of CD86 in Rapa-sTNFRI group was significantly lower than in imDC group (P＜0.05).MTT results demonstrated that T cell proliferation ability in Sirolimus group,sTNFRI group and Rapa-sTNFRI group were reduced as compared with mDC group (P＜0.05).The ELISA results revealed that the levels of IL-12 and INF-γ in rnDC group were significantly higher than other groups (P＜0.05).The levels of IL-12 and INF-γ in Rapa-sTNFRI group were significantly lower than other groups (P＜0.05).Conclusion Sirolimus combined with modified sTNFRI-IgGFc gene could synergistically inhibit maturation of DCs more effectively than Sirolimus or modified sTNFRI-IgGFc gene used alone.