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目的 :建立从人外周血诱导扩增树突状细胞 (DC)及其从凋亡癌细胞提呈抗原的方法。方法 :从正常人外周血分离获得单核细胞 ,加入50ng/mlhGM -CSF ,1000ng/mlhIL 4,隔天1次 ,共4次 ,培养第3天 ,加入γ射线照射过的胆管癌细胞 ,再继续体外培养1周后 ,用树突状细胞富集柱收集DC。结果 :DC高表达共刺激分子B7和CD1a,表面具有典型不规则突起 ,DC捕捉凋亡小体、吞噬凋亡小体 ,负载抗原的DC其激发同种异体T淋巴细胞增殖的能力进一步增强。结论 :用GM CSF加IL 4能从人外周血诱导、扩增出DC ,并可以有效提呈凋亡胆管癌细胞的抗原 ,可望成为有效的肿瘤抗原装载DC的新途径。
Objective: To establish a method for inducing dendritic cells (DCs) from human peripheral blood and presenting antigen from apoptotic cancer cells. Methods: Monocytes were isolated from normal human peripheral blood, 50ng / ml hGM-CSF and 1000ng / ml hIL 4 were added, and the cells were seeded once a day for 4 times. On the 3rd day of culture, γ-ray-irradiated cholangiocarcinoma cells After one week of in vitro culture, DCs were collected using dendritic cell enrichment columns. RESULTS: DCs expressed co-stimulatory molecules B7 and CD1a with typical irregular protrusions on the surface. DCs captured apoptotic bodies and phagocytosed apoptotic bodies. DCs with antigen-loaded DCs further enhanced their ability to stimulate proliferation of allogeneic T lymphocytes. CONCLUSION: GM-CSF plus IL-4 can induce DC from human peripheral blood and can effectively present the antigen of apoptotic cholangiocarcinoma cells, which is expected to become an effective new way to load DCs with tumor antigens.