外周血单核细胞诱导的CD123髓系树突状细胞的特性研究(精).docVIP

外周血单核细胞诱导的CD123+髓系树突状细胞的特性研究 ????????[ 11-01-28 14:46:00 ]????编辑：studa20 ???????????????????? 作者：石军, 王敏, 万云, 池田和真 【摘要】? 　　目的: 探讨体外髓系培养体系中外周血单核细胞来源的CD123+髓系树突状细胞（mDC）的生物学特性。方法: 分离健康人外周血单核细胞, 用重组的粒/单核细胞集落刺激因子（GMCSF）和白细胞介素4（IL4）将其诱导为树突状细胞（DC）。用流式细胞术（FCM）检测DC表面共刺激分子、 CD304 、 CD123和CD11C的表达, 并用间接免疫磁珠法将其中CD123+DC加以分离纯化; 激光共聚焦显微镜、 扫描电镜观察CD123+DC形态; ELISA法检测CD123+DC的IL12 分泌量; 葡聚糖吞噬试验和3HTdR渗入法分别检测CD123+DC的吞噬功能和对同种异体T细胞的刺激能力。结果: 外周血单核细胞经GMCSF和IL4诱导7 d后, 细胞表面高度表达CD86和CD11C, 中等量表达CD1a和CD123, 低表达CD83, 丧失CD14的表达, 其中, CD123和CD11C均匀分布于DC表面。免疫磁珠纯化后的CD123+DC呈现典型的不成熟DC形态, 除细胞体积较小外, 其表面突起类似于CD123-DC。CD123+DC仅微量分泌IL12, 其吞噬能力强于CD123-DC（P0.05）, 但抗原刺激功能低于后者（P0.05）。结论: GMCSF和IL4培养体系中的CD123+DC可能是DC分化发育过程中更早期的未成熟mDC, 具有独特的生物学特性。 【关键词】? 髓系培养体系; 髓系树突状细胞; CD123 　　［Abstract］AIM:? To investigate the biological characteristics of CD123+DC derived from cord blood monocytes in myeloid culture system. METHODS:? The monocytes isolated from peripheral blood were cultured with GMCSF and IL4 to induce DC for 7 days,? and the related phenotype of DC was analyzed by flow cytometery. CD123+DC were purified with magnetic cell sorting separation technique. The morphological properties of CD123+DC was observed under confocal laserscanning microscope and scan electron microscope. The concentration of IL12 secreted by DC was analyzed by ELISA. The function of endocytosis and antigen presentation was evaluated by uptaking FITCdextran and allogeneic mixed T lymphocyte reaction (MLR). RESULTS:? FACS analysis showed the high expression of CD86 and CD11C,? moderate expression of CD1a and CD123,? low levels of CD83 on DC after induction by GMCSF and IL4 for 7 days,? while the expression of CD304 was rather low. Welldistributed CD123 and CD11C were detected on DC. Purified CD123+DC showed typical morphology of immature DC with similar short cytoplasmic projections as CD123-DC. CD123+DC secreted lower level of IL12,? and exhibited more significant endocytosis and less antigen presentation function? tha