【摘要】 目的:探讨肿瘤内皮细胞抗原负载的树突状细胞(DC)诱导的细胞毒性T淋巴细胞(CTL)的特异性杀伤效应。方法:采用肿瘤细胞的培养上清诱导人脐静脉血管内皮细胞(HUVEC)增殖,制备肿瘤血管衍生的内皮细胞(TdEC)。用RTPCR检测肿瘤内皮标志物(TEM)的表达。制备TdEC的冻融抗原,负载从外周血中扩增的DC,用MTS比色法检测DC刺激自体淋巴细胞增殖的效应;用LDH法检测DC诱导的CTL的特异性杀伤效应。结果:TdEC可表达TEM1和TEM8。负载TdEC抗原的DC,可显著刺激自体淋巴细胞增殖。由其诱导的CTL对TdEC具有特异性的杀伤作用。在效靶比为20∶1和10∶1时,杀伤率分别为33%和27%,高于对照组的14%和10%。结论:TdEC抗原负载的DC,在体外可有效地诱导CTL产生,并特异性地杀伤TdEC。更多还原

【Abstract】 AIM: To study the specific cytotoxicity of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) loaded with tumor-derived endothelial cells (Td-ECs) antigen. 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 (TEM) was detected by RT-PCR. Human peripheral blood monocyte-derived DCs were loaded with Td-EC freeze-thawing antigen, and the autologous stimulation capacity of DCs was determined via mixed lymphocytes reaction employing MTS colorimetry. Specific cytolytic activity of CTLs induced by DCs was assessed by lactate dehydrogenase (LDH) assay. RESULTS: TEM1 and TEM8 were detected only on Td-ECs. DCs loaded with Td-EC antigen stimulated lymphocytes proliferation significantly. CTLs induced by DCs loaded with Td-EC antigen showed higher cytolytic activity against Td-EC than that of the control groups (33% vs 14% at E/T ratio 20, 27% vs 10% at E/T ratio 10). CONCLUSION: DC loaded with Td-EC antigen can effectively induce specific CTLs against Td-ECs in vitro.更多还原