摘要
目的:通过对体外使用Flt3-L(FL)及粒-巨噬细胞集落刺激因子(GM-CSF)诱导完全缓解期急性髓系白血病(AML-CR)患者的骨髓中单个核细胞(BMNC)分化为树突状细胞(DC)、分泌白介素(IL-12P70),干扰素-α(INF-α)的情况,及所诱导DC体外刺激自体T细胞(auto-T cells)活化增殖能力的观察,进一步了解FL在DC瘤苗制备中的作用特点。方法:分离AML-CR患者BMNC采用FL／GM-CSF单因子培养于RPMI1640完全培养基中至第8天,加入LPS刺激活化后流式细胞仪明确DC免疫表型,ELISA法测定培养液上清中IL-12P70、INF-α,收集阳性细胞体外激发增埴auto-T细胞,MTT法测试T细胞增殖情况。结果:10例AML患者的BMNC经FL／GM-CSF分别培养至第72 h均开始有细胞成簇生长,流式细胞检测显示CD1a、CD83、CD80、CD86、HLA-DR均表达明显上调,ELISA测定FL组IL-12P70,INF-α分泌量较GM-CSF组有显著增加。2组均可产生较强的T细胞增殖效应,与GM-CSF比,FL组对CD4+T细胞的增殖作用更强。结论:在体外,FL可更强诱导AML-CR患者BMNC产生DC,同时分泌大量IL-12P70及INF-α。故与GM-CSF相比,FL可能具有更佳的抗肿瘤免疫作用。
Objective：To study the differences of FL and GM-CSF expanding dendritic cells derived from AML- CR patients＇ bone marrow and the capacity of stimulating auto-T cells in vitro. Method：The bone marrow mononu- clear cells from AMLCR patients were cultured in RPMI1640 medium contained FL/GM-CSF; After 8 days, the cells were stimulated with LPS. When mature, they were analyzed for immuno-phenotype and IL-12, INF a by ELISA. Finally the DCs were used in auto-genic T cells test. Result： In all cases, a significant proportion of cells started to show DCs morphology in 3 days with different dose FL or GM-CSF. After activation, the ceils up-modulate CD1α/CD83 and CD80,CD86, HLA-DR significantly, and have potential in stimulating auto-T cells, but FL expand Th2 cells more efficient. Conclusion： Compared with GM CSF, FL could more powerfully enhance DCs from the bone marrow of AML-CR patients. These DCs have the capability to produce a great amount of IL- 12P70,INF-α and preferentially stimulate auto-T cells ex-vivo.
出处
《临床血液学杂志》
CAS
2006年第3期155-157,共3页
Journal of Clinical Hematology
