Prolongation of liver allograft survival by dendritic cells modified with NF_(-k)B decoy oligodeoxynucleotides  

作　　者：Ming-QingXu Yu-PingSuo Jian-PingGong Ming-ManZhang lu-NanYan 

机构地区：[1]DepartmentofGeneralSurgery,WestChinaHospital,SichuanUniversity,Chengdu610041,SichuanProvince,China [2]WestChinaSecondUniversityHospital,SichuanUniversity,Chengdu610041,SichuanProvince,China

出　　处：《World Journal of Gastroenterology》2004年第16期2361-2368,共8页世界胃肠病学杂志（英文版）

基　　金：Supported by the Postdoctoral Science Foundation of China,No.2003033531

摘　　要：AIM: To induce the tolerance of rat liver allograft by dendritic cells (DCs) modified with NF-κB decoy oligodeoxynucleotides (ODNs).METHODS: Bone marrow (BM)-derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL-4 to obtain immature DCs or mature DCs. GM-CSF+IL-4-propagated DCs were treated with double-strand NF-κB decoy ODNs containing two NF-κB binding sites or scrambled ODNs to ascertain whether NF-κB decoy ODNs might prevent DC maturation. GM-CSF-propagated DCs, GM-CSF+NF-κB decoy ODNs or scrambled ODNs-propagated DCs were treated with LPS for 18 h to determine whether NF-κB decoy ODNs could prevent LPS-induced IL-12 production in DCs. NF-κB binding activities, costimulatory molecule (CD40, CD80, CD86) surface expression, IL-12 protein expression and allostimulatory capacity of DCs were measured with electrophoretic mobility shift assay (ENSA),flow cytometry, Western blotting, and mixed lymphocyte reaction (MLR), respectively. GM-CSF-propagated DCs, GM-CSF+IL-4 -propagated DCs, and GM-CSF+NF-κB decoy ODNs or scrambled ODNs-propagated DCs were injected intravenously into recipient LEW rats 7 d prior to liver transplantation and immediately after liver transplantation.Histological grading of liver graft rejection was determined 7 d after liver transplantation. Expression of IL-2, IL-4 and IFN-γ, mRNA in liver graft and in recipient spleen was analyzed by semiquantitative RT-PCR. Apoptosis of liver allograft-infiltrating cells was measured with TUNEL staining.RESULTS: GM-CSF-propagated DCs, GM-CSF+NF-κB decoy ODNs-propagated DCs and GM-CSF+ scrambled ODNs-propagated DCs exhibited features of immature DCs, with similar low level of costimulatory molecule(CD40, CD80,CD86) surface expression, absence of NF-κB activation,and few allocostimulatory activities. GM-CSF+IL-4-propagated DCs displayed features of mature DCs, with high levels of costimulatory molecule (CD40, CD80, CD86) surface expression, marked NF-κB activation, and significant allocostimulatory activity. NF-κB decoyAIM:To induce the tolerance of rat liver allograft by dendritic cells (DCs) modified with NF-κB decoy oligodeoxynucleotides (ODNs). METHODS:Bone marrow (BM)-derived DCs from SD rats were propagated in the presence of GM-CSF or GM-CSF+IL-4 to obtain immature DCs or mature DCs.GM-CSF+IL-4- propagated DCs were treated with double-strand NF-κB decoy ODNs containing two NF-KB binding sites or scrambled ODNs to ascertain whether NF-KB decoy ODNs might prevent DC maturation.GM-CSF-propagated Des,GM- CSF+NF-κB decoy ODNs or scrambled ODNs-propagated Des were treated with LPS for 18 h to determine whether NF-κB decoy ODNs could prevent LPS-induced IL-12 production in Des.NF-κB binding activities,costimulatory molecule (CD40,CD80,CD86) surface expression,IL-12 protein expression and allostimulatory capacity of Des were measured with electrophoretic mobility shift assay (EMSA), flow cytometry,Western blotting,and mixed lymphocyte reaction (MLR),respectively.GM-CSF-propagated DCs,GM- CSF+IL-4 -propagated DCs,and GM-CSF+NF-κB decoy ODNs or scrambled ODNs-propagated DCs were injected intravenously into recipient LEW rats 7 d prior to liver transplantation and immediately after liver transplantation. Histological grading of liver graft rejection was determined 7 d after liver transplantation.Expression of IL-2,IL-4 and IFN-γ mRNA in liver graft and in recipient spleen was analyzed by semiquantitative RT-PCR.Apoptosis of liver allograft-infiltrating cells was measured with TUNEL staining. RESULTS:GM-CSF-propagated DCs,GM-CSF+NF-κB decoy ODNs-propagated Des and GM-CSF+ scrambled ODNs- propagated DCs exhibited features of immature DCs,with similar low level of costimulatory molecule(CD40,CD80, CD86) surface expression,absence of NF-κB activation, and few allocostimulatory activities.GM-CSF+IL-4-propagated DCs displayed features of mature DCs,with high levels of costimulatory molecule (CD40,CD80,CD86) surface expression,marked NF-κB activation,and significant allocostimulatory activity.NF-κB decoy ODNs completely abrogat

关 键 词：肝脏 同种异体移植物 树枝状细胞 NF-κB 脱氧核苷酸 手术治疗 

分 类 号：R657.3[医药卫生—外科学]