肿瘤源性血管内皮细胞RNA转染树突细胞体外诱导特异性CTL的研究  

Induction of specific CTLs using dendritic cells transfected with tumor endothelial cell RNA in vitro

作　　者：向邦德[1] 吕明德[2] 汤庆[2] 黄洁夫[2] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,广西南宁530021 [2]中山大学附属第一医院肝胆外科,广东广州510080

出　　处：《中华肿瘤防治杂志》2008年第21期1628-1631,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：高等学校博士点科研基金(20030558065)

摘　　要：目的:探讨转染肿瘤血管内皮细胞RNA的树突细胞(DC)能否诱导特异性的细胞毒性T淋巴细胞(CTL)。方法:采用肿瘤细胞的培养上清诱导人脐静脉内皮细胞(HUVEC)增殖,从而制备肿瘤源性血管内皮细胞(Td-EC)。用RT-PCR检测肿瘤内皮标志1(TEM1)在Td-EC上的表达。制备Td-EC的总RNA,转染从人外周血中扩增的未成熟DC,用流式细胞仪检测DC表型的变化,用LDH法检测DC诱导的CTL对Td-EC、HUVEC和人脐静脉内皮细胞株CRL-1730的杀伤率。结果:Td-EC特异性表达TEM1。转染RNA的DC组CD83、CD86表达率分别为(85.22±7.73)%和(88.38±8.02)%,而未成熟DC组CD83、CD86表达率分别为(51.58±9.49)%和(61.77±2.91)%,两组差异有统计学意义,P<0.05。在效靶比为20∶1、10∶1和5∶1时,CTL对Td-EC的杀伤率分别为(34.15±1.14)%、(27.02±1.31)%和(13.12±1.63)%;CTL对HUVEC杀伤率分别为(11.35±0.97)%、(8.24±0.37)%和(5.30±0.73)%;CTL对CRL-1730杀伤率分别为(13.24±1.76)%、(8.41±0.91)%和(5.31±0.94)%;CTL对Td-EC杀伤率均显著高于对HUVEC和CRL-1730的杀伤率,P<0.05。结论:转染Td-EC RNA的DC可在体外诱导CTL特异性地杀伤Td-EC。OBJECTIVE： To investigate whether dendritic cells （DCs） transfected with tumor derived endothelial cell （Td-EC） RNA could induce the specific cytotoxic T lymphocytes （CTLs）. METHODS： Td-ECs were generated from human umbilical vein endothelial cells （HUVECs） by induction with cultured supernatants of tumor cells. The expression of tumor endothelial marker 1 （TEM 1） was detected by RT PCR. Human peripheral blood monocyte-derived DCs were transfected with Td EC RNA, the expression of DC phenotype was examined with flow cytometric analysis, and the specific cytolytic activity of CTLs induced by DCs was assessed by the lactate dehydrogenase （LDH） assay. RESULTS： TEM 1 was detected only on Td ECs. The expression of CD 83 and CD 86 on DCs transfected with Td-EC RNA were （85.22±7.73）% and （88.38±8.02）%, which were much higher than those on immature DCs[（51. 58± 9.49）% and （61.77±2.91）%], P〈0.05. Thecytolyticactivities of CTLs induced by DCs transfected with Td-EC RNA against Td ECs were （34. 15±1. 14）%, （27. 02±1. 31）% and （13.12±1.63）% at the effector/target ratio 20 ： 1,10 ： 1 and 5 ： 1 respectively, which were much higher than those against HUVECs [（11. 35±0. 97）%, （8. 24±0. 37）%, （5.30±0. 73）%] and CRL-1730s [（13. 24±1. 76）%, （8.41±0.91）%, （5.31±0.94）%], P〈0. 05. CONCLUSION： DCs transefected with Td-EC RNA can effectively induce specific CTLs against tumor-derived endothelial cells in vitro.

关 键 词：肿瘤/免疫疗法 树突细胞 内皮 血管 内皮细胞 

分 类 号：R73-36[医药卫生—肿瘤]