钙载体诱导慢性髓系白血病细胞分化成树突状细胞  

Study of differentiation from CML into dendritic cell by calcium ionorphore

作　　者：周海容[1] 陈君敏[2] 陈志哲[1] 张臣青[1] 

机构地区：[1]福建医科大学附属协和医院福建省血液病研究所,福州350001 [2]福建医科大学附属第一医院,福州350001

出　　处：《中国免疫学杂志》2006年第10期948-951,共4页Chinese Journal of Immunology

基　　金：福建省自然科学基金资助项目(编号:C0210014)

摘　　要：目的:探讨用钙离子载体(Calcium ionorphore,CI)A23187诱导慢性髓系白血病(CML)病人来源的白血病细胞分化成树突状细胞(Dendritic cells,DCs)的方法和条件。方法:选择外周血白细胞计数较高的CML病人,采集外周血或骨髓液分离单个核细胞,通过两次贴壁法除去单核细胞和淋巴细胞,然后放入含或不含GM-CSF(200ng/ml)和CI(375ng/ml)的RP-MI1640培养液中培养96小时以上,通过流式细胞仪分析细胞表型、倒置显微镜和电镜观察细胞的形态变化来观察CI对白血病细胞诱导成树突状细胞的作用,初步摸索和探讨CI诱导白血病细胞转化成树突状细胞的最佳条件。结果:发现对CML病人外周血或骨髓液分离的白血病细胞加入GM-CSF(200ng/ml)、CI(375ng/ml)和含10%胎牛血清的RPMI1640培养96小时,细胞能获得典型的成熟树突状细胞的形态,CD86、CD80、CD40、CD83和HLA-DR的表达显著提高。结论:CML细胞用GM-CSF和CI联合诱导能获得成熟树突状细胞的典型形态特征和免疫表型。Objective：To explore the methods and condition in which calcium ionorphore（CI） induces the CML cells to differentiate into dendritic cells（DCs）. Methods： Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30 × 30^9 L^-1 when samples were collected,then lymphocytes and monocytes were discarded by pouting out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10% FCS（ Fetal calf serum） ,with or without CI（375 ng/ml） and GM-CSF（200 ng/ml） at 37℃ ,5% CO2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs ,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI. Results：After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80, CD86, CD40, CD86 and HLA-DR. Conelusion： CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.

关 键 词：树突状细胞 钙载体 白血病 髓系 慢性 

分 类 号：R392.12[医药卫生—免疫学]