负载肺癌抗原DC疫苗抗肿瘤免疫实验

Antitumor Immune Experiment of Dendritic Cell Vaccine Loaded with Lung Cancer Antigen

目的探讨负载人肺癌细胞株A549冻融抗原DC疫苗体外抗肿瘤免疫效应。方法自脐血分离单核细胞,经rhGM-CSF、rhIL-4和rhTNF-α诱导树突状细胞(DC),负载人肺癌细胞株A549冻融抗原、LPS诱导成熟,利用免疫磁珠分选获得功能性肺癌DC疫苗(UbDC/AgL),ELISA测定UbDC/AgL培养上清中细胞因子浓度、LDH法测定特异性细胞杀伤(CTL)活性及MTT法检测自身混合淋巴细胞反应(AMLR)。结果UbDC/AgL细胞组IL-1、IL-12、IL-6和IFN-β含量明显高于UbDC/Ag和UbDC组(P<0.01);UbDC/AgL疫苗细胞能有效诱导肿瘤特异性CTL活性,对A549细胞有特异性杀伤作用(P<0.01),并能有效促进淋巴细胞增殖。结论负载肿瘤冻融抗原DC疫苗,能有效激活T淋巴细胞进而诱导肿瘤特异性杀伤效应。

Objective To investigate the antitumor effect of dendritic cell vaccine loaded with the human lung cancer associated antigens. Methods The monocytes were separated from human umbilicus cord blood cells and cultured for dendritic cells （UbDC） with rhGM-CSF, rhIL-4 and rhTNF-α in RPMI 1640 medium. The human lung cancer cells A549 cytolysis antigen and LPS were added into the medium. Successively for loading the A549 antigen on UbDC and promoting DCs mature. The mature DC vaccine loaded with A549 cytolysis antigen（UbDC/AgL） was sorted by magnetic beads. MTT assay was employed to test the auto mix lymphocyte reaction （AMLR）. LDH release assay was carried out to assess the killing ability of CTL cells against A549 lung cancer cells. The level of human IL-1, IL-12, IL-6 and TNF-β in the culture supernatant was determined by ELISA. Results The level of human IL-1, IL-12, IL-6 and TNF-β in the culture supernatant of UbDC/AgL were higher than that of UbDC/Ag and UbDC separately（P〈0. 01）. The special CTL cells had significant cytotoxicity against A549 cells （P〈0. 01） and inducemented lymphopoiesis availability. Conclusion The dendritic cell vaccine loaded with tumor cytolysis antigen can activate naive T lymphocyte and induce specific antitumor immune response efficiency.