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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。
Objective: To investigate the specific killing effect of dendritic cells (DCs) -induced cytotoxic T lymphocytes (CTLs) loaded with tumor endothelial cells. Methods: The proliferation of human umbilical vein endothelial cells (HUVECs) was induced by culture supernatant of tumor cells to prepare tumor vascular derived endothelial cells (TdECs). RTPCR was used to detect the expression of tumor endothelial markers (TEMs). The TdEC frozen-thawed antigen was prepared and DCs amplified from peripheral blood were loaded. The effect of DCs on the proliferation of autologous lymphocytes was detected by MTS colorimetric assay. The specific killing effect of CTL induced by DCs was detected by LDH assay. Results: TdEC can express TEM1 and TEM8. DCs loaded with TdEC antigen can significantly stimulate autologous lymphocyte proliferation. The CTL induced by it has a specific killing effect on TdEC. The killing ratios were 33% and 27%, respectively, higher than the control group by 14% and 10% at 20: 1 and 10: 1, respectively. Conclusion: TdEC-loaded DC can effectively induce CTL production in vitro and kill TdEC specifically.