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目的 :证实树突状细胞 (Dendritic cells,DCs)可通过吞噬凋亡小体获取抗原物质 ,探讨其在肿瘤免疫治疗中的意义。方法 :利用吸附单克隆抗体 -磁珠分离系统 (MACS) ,从人脐血中分离出 CD34+ 细胞 ,体外以重组 h GM- CSF+ h SCF+ h TNF-α+ h FL诱导培养 DCs,光镜观察诱生细胞的形态 ,流式细胞术 (FACS)检测其表型。电镜观察和流式细胞仪检测三氧化二砷 (As2 O3 )诱导凋亡的 HL - 6 0细胞 ,将凋亡的 HL- 6 0细胞与培养早期的 DCs共育 4～ 8h,电镜观察 DCs吞噬凋亡小体的现象。结果 :CD34+细胞经诱生后生成了大量具有典型形态和表型的成熟 DCs;电镜下见凋亡的 HL- 6 0细胞内凋亡小体形成 ,流式细胞仪检测 DNA含量 ,见亚二倍体峰形成。凋亡细胞与 DCs共育后 ,电镜下见 DCs吞噬凋亡小体的现象。结论 :DCs可通过吞噬凋亡小体摄取抗原物质。
Objective: To demonstrate that dendritic cells (DCs) can acquire antigenic substances through phagocytosis of apoptotic bodies and explore their significance in tumor immunotherapy. Methods: CD34 + cells were isolated from human umbilical cord blood by adsorption of monoclonal antibody-magnetic bead separation system (MACS). DCs were induced by recombinant h GM-CSF + h SCF + h TNF-α + h FL in vitro. The morphology of green cells, flow cytometry (FACS) to detect the phenotype. Electron microscopy and flow cytometry were used to detect the apoptosis of HL - 60 cells induced by As 2 O 3. The apoptotic HL - 60 cells were incubated with early DCs for 4 ~ 8 hours. The apoptotic HL - 60 cells were observed by electron microscopy Body phenomenon. Results: A large number of mature DCs with typical morphology and phenotype were generated after induced by CD34 + cells. Apoptotic bodies were observed in apoptotic HL-60 cells under electron microscope. DNA content was detected by flow cytometry, Ploidy peak formation. Apoptotic cells co-cultured with DCs, under the electron microscope, DCs engulf apoptotic bodies phenomenon. Conclusion: DCs can ingest the antigenic substance by phagocytosis of apoptotic bodies.