Immunological tolerance induced by Sirolimus combine with modified sTNFRI-IgGFc gene in rat dendritic cells

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. Key words: Rats; Dendritic cells; Rapamycin; Gene transfection; Immune tolerance