摘要
目的观察转化生长因子-β1(TGF-β1)基因转染未成熟树突状细胞(imDC)对大鼠肝移植免疫耐受的诱导作用。方法利用大鼠骨髓细胞诱导培养imDC,以脂质体介导的pIRES2-EGFP—hTGF—Bl转染imDC,于移植前5d输注受体Lewis大鼠体内,移植后分别于3、7、10d抽血检测肝功能[总胆红素(TBIL)和谷丙转氨酶(ALT)]、酶联免疫吸附试验(ELISA)法检测白细胞介素-12(IL-12)水平、肝组织苏木素-伊红(HE)染色急性排斥反应病理评分、原位末端转移酶标记(TUNEL)法检测肝脏淋巴细胞的凋亡及观察各组大鼠肝移植后的生存时间。结果TGF-β1组移植后肝功能(TBIL和ALT)优于各对照组(P〈0.05);TGF—β1组移植后3、7d血清IL-12浓度分别为71.03±10.70、80.88±14.23均低于各对照组(P〈0.05);TGF-β1组移植后7d出现交界性排斥反应,移植10d后出现轻度排斥反应(P〈0.05);TGF-β1基因转染组移植后3、7、10d汇管区淋巴细胞凋亡指数分别为6.75±1.93、14.00±2.19、18.25±1.38,较各对照组均明显增高(P〈0.05);TGF-β1组大鼠移植后中位生存期为58d,明显长于各对照组。结论TGF—β1基因改造的imDC能诱导大鼠移植免疫耐受,在诱导器官移植耐受中有良好时应用前景。
Objective To assess the effect of immature dendritic cells (imDCs) modificated with transforming growth factor-β1 ( TGF-β1 ) on immune tolerance in a rat liver transplantation. Methods The animals were divided into allogenic transplantation, imDC, mature dendritic cell (mDC) , plRES2- EGFP, and TGF-β1. Rat bone marrow-derived imDCs were obtained after primary culture of bone marrow cells in RPMI 1640 medium with different cytokines. ImDCs transfected with plasmid vectors encoding TGF-β1 were injected into the allogenie Lewis rat 5 days before liver transplantation. Liver functions [ total bilirubin (TBIL) and alanine aminotransferase (ALT) ], interleukin (IL)-12 level in the blood, liver pa- thology by hematoxylin and eosin (HE). staining, acute liver rejection grades valued by Baff standard, apoptosis of lymphocyte in recipient liver analyzed by Tunnel staining, and survival time were observed 3 d, 7 d and 10 d post liver transplantation respectively. Results TBIL and ALT were much lighter in Group TGF-β1 than control groups(P 〈0.05). The level of IL-12 [ (71.03±10. 70), (80. 88±14. 23) μg/L respectively] in 3,7 d after liver transplantation is much lower in Group TGF-β1 than control groups (P 〈 0. 05). Apoptosis of T cells in 3, 7, 10 d after liver transplantation (6. 75 _+ 1.93, 14. 00 ±2. 19, 18.25±1.38) is much more in group TGF-β1 than control groups. Light acute rejection was observed in TGF-β1 group in 10 d after liver transplantation( P 〈 0.05 ). The medium survival time is about 58 day, which is longer than in control groups ( P 〈 0. 05 ). Conclusion ImDCs modificated with TGF-β1 can enhance the immue tolerogenicity in a rat liver transplantation, it might be a promising therapy for donor-specific toler- ance after organ transplantation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第5期873-876,共4页
Chinese Journal of Experimental Surgery
基金
福建省自然科学基金资助项目(2009D066)
福建医科大学校重大课题项目(09ZD007)
关键词
树突状细胞
肝移植
免疫耐受
转化生长因子-Β1基因
Dendritic cells
Immue tolerance
Liver transplantation
Transforming growth factor-β1 gene