人PBMC和脐血CD34^+细胞诱导DC2的实验方法研究  

Establish a method for Inducing DC2 via CD34^+ hematopoietic stem cells from Human cord blood or PBMC

作　　者：王金湖[1] 邵启祥[2] 阴晴[2] 王胜军[2] 潘湘涛[1] 顾超[1] 许化溪[2] 

机构地区：[1]太仓市人民医院检验科,江苏215400 [2]江苏大学医学技术学院微生物学和免疫学教研室

出　　处：《中国交通医学杂志》2006年第2期167-170,共4页Chinese Medical JOurnal of Communications

基　　金：江苏省科委应用基础项目(BJ99041);江苏省教委立项项目(98KDJ31004);江苏省科委社会发展立项项目(SS20115)

摘　　要：目的:探讨从人外周血单个核细胞(PBMC)和脐血CD34+造血干细胞(HSC)诱导DC2方法。方法:分别用rhIL-3(10ng/ml)+rhG-CSF(100ng/ml)和rhFlt-3L(100ng/ml)+rhIL-3(10ng/ml)+rhG-CSF(100ng/ml)诱导人PBMC和脐血CD34+HSC向DC2分化,并采用流式细胞仪分析细胞的表型,同时观察rhTNF-α和人CD40单抗诱导DC2分化成熟的作用。结果:rhG-CSFh+rhIL-3未能较好地从PBMC中诱导出DC2,但是从富集的脐血CD34+HSC用rhFlt-3L+rhIL-3+rhG-CSF诱导,可以获得相对较多的DC2,并有较高的扩增效率,但离临床研究和应用还有一定距离,因此DC2的诱导方法仍需进一步优化。人CD40单抗和rhTNF-α一样,能有效地诱导DC2分化成熟。结论:体外联合多种细胞因子从CD34+HSC成功地诱导出DC2,抗人CD40单抗有利于DC2的活化成熟。Objective：To investigate the conditions of inducing DC2 via human peripheral blood mononuclear cells （PBMC） or CD34^＋ hematopoietic stem cells （HSC） enriched from human cord blood and evaluate the effect of anti-CD40 McAb in inducing DC2 maturation. Methods：PBMC was isolated from blood and was CD34^＋ HSC enriched from cord blood of healthy volunteers respectively. The pre-DC from PBMC or CD34^＋ HSC were generated with rhIL-3（10ng/ml）, rhG-CSF（100ng/ml） or rhIL-3（10ng/ml）, rhG-CSF （100ng/ml） and rhFh-3L（100ng/ml）, and then was the membrane surface mark of DC2 with FCM identified. Results：CD123^＋ DC2 were successfully induced from CD34^＋ cord blood HSC with rhFh-3L,rhIL-3 and rhG-CSF, but were unsuccessfully induced from PBMC with rhIL-3 and rhG-CSF in vitro. Furthermore, DC2 were induced to maturation by anti-human CD40 McAb.Conclusions：DC2 can be induced from CD34^＋ cord blood HSC with rhFlt-3L,rhIL-3 and rhG-CSF and be induced to maturation by anti-human CD40 McAb in vitro.

关 键 词：CD34^+ 造血干细胞 rhFh-3L RHIL-3 RHG-CSF DC2 

分 类 号：R392.3[医药卫生—免疫学]