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7d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(P<0.05),但抗原刺激功能低于后者(P<0.05)。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。
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 than that of CD123-DC（P〈0.05）.CONCLUSION：CD123＋DC induced with GM-CSF and IL-4 might be more early phase of immature myeloid DC along with the DC differentiation,with special biological characteristics.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2009年第3期204-207,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
上海市青年科技启明星课题(02QB14035)
上海市卫生局青年课题(024Y14)
