中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2010年
8期
488-491
,共4页
RNA干扰%共刺激通路%树突状细胞CD40%心脏移植
RNA榦擾%共刺激通路%樹突狀細胞CD40%心髒移植
RNA간우%공자격통로%수돌상세포CD40%심장이식
RNA interference%Costimulatory pathway%Dendritic cell%CD40%Heart transplantation
目的 探讨慢病毒介导RNA干扰(RNAi)技术阻断CD40/CD40L共刺激通路对小鼠移植心存活时间的影响.方法 以针对小鼠CD40基因的RNAi慢病毒载体在体外感染供者骨髓来源的树突状细胞(DC),制备低表达CD40的耐受性DC(Tol-DC).荧光实时定量聚合酶链反应及流式细胞术检测DC感染前后CD40 mRNA及DC表面抗原CD40、CD11c和MHCⅡ的表达.建立小鼠异位腹腔心脏移植模型.在小鼠异位心脏移植前7 d,经静脉给受者输注体外制备的低表达CD40的Tol-EC(慢病毒感染DC注射组),并设置单纯移植对照组和未感染DC注射组作为对照.观察各组移植心的存活时间,评定各组术后第7天移植心排斥反应的病理分级.结果 CD40-RNAi慢病毒载体在体外感染DC 48 h后,CD40 mRNA表达明显受到抑制,抑制率为80.9%;CD40表达明显下降,由(74.37±4.08)%降至(40.07±4.03)%(P＜0.05).单纯移植对照组、未感染DC注射组和慢病毒感染DC注射组移植心存活时间分别为:(8±2)d、(9±1)d和(14±4)d,慢病毒感染DC注射组移植心存活时间明显延长,并且移植心排斥反应病理分级显著降低,差异有统计学意义(P＜0.05).结论 阻断CD40/C40L共刺激通路可抑制异系T淋巴细胞的活化,从而抑制急性排斥反应,延长小鼠移植心的存活时间.
目的 探討慢病毒介導RNA榦擾(RNAi)技術阻斷CD40/CD40L共刺激通路對小鼠移植心存活時間的影響.方法 以針對小鼠CD40基因的RNAi慢病毒載體在體外感染供者骨髓來源的樹突狀細胞(DC),製備低錶達CD40的耐受性DC(Tol-DC).熒光實時定量聚閤酶鏈反應及流式細胞術檢測DC感染前後CD40 mRNA及DC錶麵抗原CD40、CD11c和MHCⅡ的錶達.建立小鼠異位腹腔心髒移植模型.在小鼠異位心髒移植前7 d,經靜脈給受者輸註體外製備的低錶達CD40的Tol-EC(慢病毒感染DC註射組),併設置單純移植對照組和未感染DC註射組作為對照.觀察各組移植心的存活時間,評定各組術後第7天移植心排斥反應的病理分級.結果 CD40-RNAi慢病毒載體在體外感染DC 48 h後,CD40 mRNA錶達明顯受到抑製,抑製率為80.9%;CD40錶達明顯下降,由(74.37±4.08)%降至(40.07±4.03)%(P＜0.05).單純移植對照組、未感染DC註射組和慢病毒感染DC註射組移植心存活時間分彆為:(8±2)d、(9±1)d和(14±4)d,慢病毒感染DC註射組移植心存活時間明顯延長,併且移植心排斥反應病理分級顯著降低,差異有統計學意義(P＜0.05).結論 阻斷CD40/C40L共刺激通路可抑製異繫T淋巴細胞的活化,從而抑製急性排斥反應,延長小鼠移植心的存活時間.
목적 탐토만병독개도RNA간우(RNAi)기술조단CD40/CD40L공자격통로대소서이식심존활시간적영향.방법 이침대소서CD40기인적RNAi만병독재체재체외감염공자골수래원적수돌상세포(DC),제비저표체CD40적내수성DC(Tol-DC).형광실시정량취합매련반응급류식세포술검측DC감염전후CD40 mRNA급DC표면항원CD40、CD11c화MHCⅡ적표체.건립소서이위복강심장이식모형.재소서이위심장이식전7 d,경정맥급수자수주체외제비적저표체CD40적Tol-EC(만병독감염DC주사조),병설치단순이식대조조화미감염DC주사조작위대조.관찰각조이식심적존활시간,평정각조술후제7천이식심배척반응적병리분급.결과 CD40-RNAi만병독재체재체외감염DC 48 h후,CD40 mRNA표체명현수도억제,억제솔위80.9%;CD40표체명현하강,유(74.37±4.08)%강지(40.07±4.03)%(P＜0.05).단순이식대조조、미감염DC주사조화만병독감염DC주사조이식심존활시간분별위:(8±2)d、(9±1)d화(14±4)d,만병독감염DC주사조이식심존활시간명현연장,병차이식심배척반응병리분급현저강저,차이유통계학의의(P＜0.05).결론 조단CD40/C40L공자격통로가억제이계T림파세포적활화,종이억제급성배척반응,연장소서이식심적존활시간.
Objective To investigate the effect of blocking CD40/CD40L costimulatory pathway by the lentiviral vector-mediated RNA interference on the survival of mouse cardiac allograft. Methods Mouse bone marrow-derived dendritic cells (DCs) were infected by CD40-RNAi lentiviral vector in vitro, and tolerogenic DCs (Tol-DCs) with decreased CD40 expression were prepared. Fluorescence real-time quantitative PCR and flow cytometry were used to analyze the expression of CD40 mRNA and DC surface antigens CD40, CD11c, MHC Ⅱ before and after infection. Mouse model of heterotropic abdominal heart transplantation was established. Seven days prior to heart transplantation, Tol-DCs with decreased CD40 expression were transfused into recipient mice intravenously (lentivirus infected DC group). Control group and non-infected DC group were assigned simultaneously. The survival of cardiac allograft was monitored and pathological grade of acute rejection 7 days after heterotropic abdominal heart transplantation was determined. Results The transcription of CD40 mRNA of DCs was down-regulated significantly at 48 h after CD40-RNAi lentiviral vector infection, and the inhibition rate was 80. 9%. The expression of CD40 protein was also significantly decreased as compared with control group (40. 07% ± 4. 03% ) ( P ＜ 0. 05 ).Compared to control group (8 ± 2 days) and non-infected DC group (9 ± 1 days), the survival time of cardiac allograft in CD40-RNAi lentivirus infected DC group (14 ± 4 days) was significantly prolonged (P＜ 0. 05 ), and the pathological grade of acute rejection decreased significantly ( P ＜ 0. 05 ).Conclusion Blocking CD40/CD40L costimulatory pathway could hamper the activation of allogeneic T lymphocyte, inhibit the acute rejection and prolong the survival of mouse cardiac allograft.
