Shi Jun, Wang Min, Wan Yun, Ikeda Kazuma
Department of Hematology, Sixth People's Hospital of Shanghai Jiaotong University, Shanghai, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Mar;25(3):204-7.
To investigate the biological characteristics of CD123(+)DC derived from cord blood monocytes in myeloid culture system.
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).
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).
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.
研究髓系培养体系中脐血单核细胞来源的CD123(+)树突状细胞（DC）的生物学特性。
将从外周血分离的单核细胞用粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-4（IL-4）培养7天以诱导DC，通过流式细胞术分析DC的相关表型。用磁珠细胞分选分离技术纯化CD123(+)DC。在共聚焦激光扫描显微镜和扫描电子显微镜下观察CD123(+)DC的形态学特性。用酶联免疫吸附测定（ELISA）分析DC分泌的白细胞介素-12（IL-12）的浓度。通过摄取异硫氰酸荧光素（FITC）-葡聚糖和同种异体混合T淋巴细胞反应（MLR）评估内吞作用和抗原呈递功能。
流式细胞术分析显示，GM-CSF和IL-4诱导7天后，DC上CD86和CD11C高表达，CD1a和CD123中度表达，CD83低表达，而CD304表达相当低。在DC上检测到CD123和CD11C分布良好。纯化的CD123(+)DC显示出未成熟DC的典型形态，与CD123(-)DC一样具有类似的短细胞质突起。CD123(+)DC分泌较低水平的IL-12，并且与CD123(-)DC相比表现出更显著的内吞作用和较弱的抗原呈递功能（P<0.05）。
用GM-CSF和IL-4诱导的CD123(+)DC可能是随着DC分化的未成熟髓系DC的更早阶段，具有特殊的生物学特性。
