慢性髓性白血病来源的树突状细胞的体外扩增培养  

Expansion of dendritic cells derived from patients with chronic myeloid leukemia in vitro

作　　者：吉蕾[1] 邢佩霓[2] 魏绪仓[2] 李梅生[2] 王彤[2] 

机构地区：[1]西安交通大学第二医院血液科,陕西西安710004 [2]陕西省人民医院血液科,陕西西安710068

出　　处：《第四军医大学学报》2003年第11期1010-1013,共4页Journal of the Fourth Military Medical University

摘　　要：目的 :探讨慢性髓性白血病 (CML)来源的树突状细胞 (DC)的体外扩增及其功能 .方法 :采用两步法 ,首先将慢性髓性白血病骨髓或外周血CD34+ 细胞与rhFlt3 L和rhTPO共育 7d,再用rhGM CSF ,rhTNF α及rhIL 4诱导培养1 4d ;同时以rhGM CSF ,rhTNF α及rhIL 4直接诱导体系作对照 .获得的DC通过免疫表型检测、电镜分析及染色体鉴定 ,并用MTT法检测其刺激T细胞增殖能力及T细胞对CML细胞的杀伤作用 .结果 :两步法培养 2 1d ,CML的CD34+ 细胞扩增 (76 . 5 6± 5 . 1 7)倍 ,DC产率 (39. 1 0± 8. 0 3) % ,直接诱导产生DC扩增倍数及比例均低于两步法 (P <0 . 0 1 ) ,且此DC能刺激自体或异体T细胞增殖进而杀伤CML细胞 .结论 :两步法体外扩增培养可明显提高DC前体总数及产率 ,优于直接诱导培养 ;AIM: To investigate the expansion and the function of dendritic cells derived from chronic myeloid leukemia cells. METHODS: DC was cultured in two steps: first CD34 + or mononuclear cells isolated from CML patients' bone marrow and peripheral blood with rhFlt3 L and rhTPO was cultured for 7 days and then they were coincubated with rhGM CSF, rhTNF and rhIL 4 to induce DCs for 14 days. A directly DCs induced system was established for comparison. DCs were identified by immunophenotype, chromsome and electric microscopy analysis. The function of stimulating T cells proliferation and cytotoxicity of CML cells was confirmed by MTT assay. RESULTS: After the two step culture, CD34 + cells had a total cell number with (76.56±5.17) fold expansion and the yield of (39.10± 8.03 )% DCs respectively for 21 days, both of which were higher than those by the system without expansion culture ( P <0.01). This DCs stimulated T cells to?proliferate and finally killed the leukemia cells. CONCLUSION: Two step culture can obviously improve the cell number and the production of DCs, better than directly inducing method. DCs derived from CML cells can induce the generation of anti leukemia immunization.

关 键 词：树突状细胞 白血病 髓性 慢性 扩增 细胞毒性T淋巴细胞 

分 类 号：Q461[生物学—生理学] Q253