慢性粒细胞白血病树突状细胞的体外无血清培养及杀瘤效应

Serum-free culture of dendritic cells from the patients of chronic myeloid leukemia in vitro and estimation of their cytotoxicity

目的 建立慢性粒细胞白血病 (CML)树突状细胞 (DCs)体外无血清培养体系。方法 小牛血清 (FCS)、无血清 (SFM)及自体血清分别培养CML病人CD34 + 细胞或单个核细胞 (MNCs) ,以SCF、GM CSF、TNF α、IL 4 (A)与GM CSF、TNF α、IL 4 (B)两组不同的细胞因子作对照。结果 CML DCsHLA DR高表达 ,CD80、CD83和CD86表达比例不高 ,异体混合淋巴细胞反应能力不强。SFM同FCS体系相比无显著差异。 5例CML DCsPh染色体比例与培养前MNCsPh染色体比例基本吻合 ;同时以CML DCs同自身T细胞、自体Ph+ MNCs共孵育 ,测其杀伤率为 (38.5± 6 .5 ) %。结论 CML DCs携带Ph染色体 ,能刺激自体T细胞杀伤自身白血病细胞 ,但需进一步改善其功能。

Objective To establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells. Methods Fetal calf serum (FCS), serum-free medium (SFM) and autologous serum were used for culture of DCs. The usage of cytokines was classified into group A (SCF, GM-CSF, TNF-α and IL-4) and group B (GM-CSF, TNF-α and IL-4). Results The results of SFM had no significant difference from those of FCS medium, but the culture system of autologous serum was inferior to them. The expression of HLA-DR on the surface of CML-DCs was notable (>50%), but the expression of CD83 and the costimulatory molecules CD80&CD86 were not noticeable (<50%). Although CD1a +/CD14 - DCs were potent stimulators of allogeneic lymphocytes, expansions of T cells from normal volunteers were not significant. At days 10～12, CD1a + cells from five patients were studied by displaying G banding and Ph-positive cells in these population were 100%, 100%, 100%, 98% and 60%, respectively. At an effector:target ratio of 40∶1, 32% to 45% cytotoxicity was noted with DC-stimulated T cells against autologous Ph + MNCs. Conclusion DCs from CML patients can be derived from the malignant clone and these malignant DCs can induce antileukemic reactivity in autologous T lymphocytes without the necessity of additional exogenous antigens. However, functions of CML-DCs need to be further improved.