G-CSF动员的供体外周血树突状细胞免疫生物学特性的研究  

Immunobiologic properties of dendritic cells derived from G-CSF mobilized peripheral blood

作　　者：汪健[1] 王保龙[2] 叶书来[2] 陈家萍[2] 王丹[2] 廖艳秋[2] 汪帮来[2] 徐军[2] 

机构地区：[1]安徽医科大学附属省立医院血液科,安徽合肥230001 [2]安徽医科大学附属省立医院输血科,安徽合肥230001

出　　处：《中国输血杂志》2009年第4期264-268,共5页Chinese Journal of Blood Transfusion

基　　金：安徽省自然科学基金资助课题(编号:070413261X);安徽省高等学校省级科研重点项目资助课题(KJ2007A031)

摘　　要：目的探讨从以G-CSF动员、CS-3000 plus血细胞分离机采集的供者外周血中诱导产生不同特性树突状细胞(DC)的可行性及DC的生物学特性。方法实验组为用CS-3000 plus血细胞分离机采集的5名造血干细胞捐献者的单核细胞(PBMCs),PBMCs培养贴壁后,经含IL-4、GM-CSF的无血清培养基诱导培养7 d,进一步分为4组:未刺激、肿瘤坏死因子α(TNF-α)、IL-10+TNF-α、抗胸腺球蛋白(ATG)+TNF-α组;对照组为5名健康献血者周血,亦按上述条件分组,每组复孔。后2组分别为IL-10、ATG诱导1 d后再加入TNF-α诱导1 d;分别测定各组DC的免疫表型、IL-12(P70)分泌、抗原吞噬以及异基因T淋巴细胞刺激反应。结果诱导培养9 d后可获得非成熟DC(iDC),单纯TNF-α刺激后,HLA-DR、CD11c、CD40、CD80的表达均明显增高,IL-12分泌增加,对异基因T淋巴细胞刺激反应性增强;而IL-10可以使CD1a表达上调,IL-10和ATG均抑制HLA-DR、CD11c、CD40、CD80的表达;与诱导的成熟DC相比,IL-10、ATG诱导DC的IL-12(P70)的分泌、对异基因淋巴细胞刺激反应性均减低,抗原摄取能力增强,但ATG、IL-10诱导的DC生物学特性不完全一致;动员的供体与对照健康献血者诱导的DC在产率,HLA-DR、CD11c表达,成熟诱导后CD11c、CD40、CD80表达,IL-12分泌,异基因T细胞刺激反应性等相比均明显减低。结论G-CSF动员的供体单核细胞PBMCs通过常规诱导可以产生iDC,TNF-α可诱导其分化成熟,IL-10、ATG可以使其获得致耐受的特性;与健康献血者相比,G-CSF动员的供体DC表型和功能部分减低,;虽然动员的供体DC产率不及健康献血者,但血细胞分离机可以可采集到大量的单个核细胞(PBMCs),因此G-CSF动员的外周血有望成为不同DC的重要来源途径。Objective To explore the possibility of inducing DCs derived from donors＇ PBMCs mobilized by G-CSF and collected with CS-3000plus, and to analyze their immunobiologic properties. Methods The adherent monocytes were propagated in serum-free medium combined with GM-CSF and IL-4 for 7 days and then divided into four groups ： no stimulation group, TNF-α group, IL-10 plus TNF-α group and ATG plus TNF-α group. The peripheral blood of healthy blood donors served as control. There were five samples in each group, and each sample was tested in double. Ceils were indueed with IL-10 or ATG for one day, and then induced with TNF-α for another day. The immunophenotype, secretion of IL-12 （ P70）, antigen phagoeytosis and ailogeneie stimulation response were examined. Results Immature DCs after 9-day induction were obtained. The expression of HLA-DR, CD11c, CD40, CD80, secretion of IL-12, and the activity of allogeneic stimulation response were all increased after induction with TNF-α .IL-10 increased the expression of CDIa. Both IL-10 and AT（; inhibited the expression of HLA-DR, CD11c, CD40 and CD80. Compared to mature DCs, IL-12 secretion and activity of allogeneic stimulus response of DCs induced by IL-10 group and ATG group were decreased, while the function of antigen uptake was enhanced. However, the biological properties of DCs induced by IL-10 and AT（; were different from each other. Compared with the DCs derived from the controls, the yield of DC, the expression of HLA-DR and CD11c, the expression of CD11c, CD40 and CD80 after induction, the secretion of IL-12, and the activity of aUogeneic stimulus response all were decreased in DCs derived from G-CSF mobilized donors. Conclusions Immature DCs could be generated from monoeytes via routine method. The mature DCs could be induced by TNF-α, and then acquire the tolerogenic properties with IL-10 or ATG The immunophenotype and function of DCs from G-CSF mobilized donors were inferior to those from healthy blood donors. Although the yield of DCs was lower

关 键 词：树突状细胞 G—CSF动员 IL-10 ATG TNF-α 混合淋巴细胞反应 

分 类 号：R446.113[医药卫生—诊断学]