CAJ | 학술논문

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-β1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10 d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58 d,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的应用前景.
목적 관찰전화생장인자-β1 (TGF-β1)기인전염미성숙수돌상세포(imDC)대대서간이식면역내수적유도작용.방법 이용대서골수세포유도배양imDC,이지질체개도적pIRES2-EGFP-hTGF-β1전염imDC,우이식전5d수주수체Lewis대서체내,이식후분별우3、7、10d추혈검측간공능[총담홍소( TBIL)화곡병전안매(ALT)]、매련면역흡부시험(ELISA)법검측백세포개소-12(IL-12)수평、간조직소목소-이홍(HE)염색급성배척반응병리평분、원위말단전이매표기(TUNEL)법검측간장림파세포적조망급관찰각조대서간이식후적생존시간.결과 TGF-β1조이식후간공능(TBIL화ALT)우우각대조조(P＜0.05)；TGF-β1조이식후3、7d혈청IL-12농도분별위71.03±10.70、80.88±14.23균저우각대조조(P＜0.05)；TGF-β1조이식후7d출현교계성배척반응,이식10 d후출현경도배척반응(P＜0.05)；TGF-β1기인전염조이식후3、7、10d회관구림파세포조망지수분별위6.75±1.93、14.00±2.19、18.25±1.38,교각대조조균명현증고(P＜0.05)；TGF-β1조대서이식후중위생존기위58 d,명현장우각대조조.결론 TGF-β1기인개조적imDC능유도대서이식면역내수,재유도기관이식내수중유량호적응용전경.
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 ),pIRES2-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 allogenic Lewis rat 5 days before liver transplantation.Liver functions [ total bilirubin (TBIL) and alanine aminotransferase (ALT) ],interleukin (IL)-12 level in the blood,liver pathology 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 tolerance after organ transplantation.