用含有特定细胞因子的无血清培养基培养慢性乙型肝炎病人的树突状细胞  

Culture of dendritic cells originated from peripheral blood in patients with chronic hepatitis B by serum-free media containing specific cytokines

作　　者：李若冰[1] 陈红松[1] 费然[1] 王松霞[1] 范春蕾[1] 魏来[1] 王宇[1] 

机构地区：[1]北京大学人民医院肝病研究所,北京100044

出　　处：《免疫学杂志》2002年第6期459-462,464,共5页Immunological Journal

基　　金：国家"973" (G1 9990 541 0 6);国家"863"(1 0 2 1 2 0 2 0 7);北京市"二四八"工程 (9540 2 540 0和 9540 2 0 70 0 )资助项目

摘　　要：目的　体外扩增慢性乙肝病人的树突状细胞 (dendriticcell,DC) ,从细胞表型和功能上鉴定和研究。方法　用含GM CSF和IL 4的无血清培养基AIM V体外培养慢性乙肝病人外周血单个核细胞 ,获得树突状细胞。流式细胞仪检测细胞表型 ,IL 1 2ELISA试剂盒检测DC分泌IL 1 2的水平 ,并观察加入细胞因子TNF α后对DC培养的影响。结果　慢性乙肝病人的外周血单个核细胞用AIM V培养及细胞因子诱导后 ,经贴壁法纯化 1 0 0mL可获得 0 .5× 1 0 7～ 1 .5× 1 0 7成熟的具有典型形态的DC ,加入TNF α后 ,CD83阳性占 60 .80 % ,明显高于未加组 (P <0 .0 5) ,IL 1 2分泌较未加TNF α组增高近 1 0倍。结论　①慢性乙肝病人的DC可用AIM V无血清培养基及特定的细胞因子诱导在体外大量获得 ,TNF α是诱导DC成熟的重要的细胞因子。②典型的细胞形态和CD1 4 -、HLA DRhigh+、CD86high+的细胞表面分子特征可作为临床上快速鉴定培养DC的标志。Objective To culture and identify dendritic cells (DC) of patients with chronic hepatitis B, then study their cell phenotype and functions.Methods Peripheral blood monocytes (PBMC) were isolated from patients with chronic hepatitis B. DCs were generated by culturing PBMC with AIM V including GM CSF and IL 4 in vitro . The changes of cell phenotype analysis on different days with or without TNF α were tested by immuno labelling and flow cytometry. IL 12 ELISA kit was applied to investigate IL 12 secretion of DCs.Results DCs of patients with typical morphology were harvested successfully in vitro . When adding TNF α, IL 12 secretion of these DCs was upregulated, meanwhile the CD83 expression increased.Conclusions ①DCs of patients are cultured successfully with AIM V when induced by GM CSF, IL 4 and TNF α in vitro . TNF α is an important cytokine for inducing maturation of DC. ②Typical cell morphology and HLA DR,CD86 expression can be used as the markers for fast identification of DC in clinical practice.

关 键 词：树突状细胞 病毒性乙型肝炎 无血清培养基 细胞因子 流式细胞仪 

分 类 号：R512.62[医药卫生—内科学]