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目的研究小鼠骨髓细胞来源的树突状细胞对细菌颗粒抗原交叉递呈的动力学特点。方法取小鼠骨髓细胞,采用GM-CSF诱导培养。用倒置显微镜观察培养过程中细胞形态,用流式细胞仪进行细胞纯度和表型测定。采用第7天未成熟的DC细胞进行细菌抗原的交叉递呈能力检测和动力学分析。结果小鼠骨髓细胞经GM-CSF诱导培养,第7天获得大量具有典型形态和免疫表型的树突状细胞,并且在细菌脂多糖刺激后,其表型也发生特征性变化表现在共刺激分子CD80由刺激前的27.7%上调到71.6%,CD86从刺激前的36.0%上调到80.3%,DEC205从刺激前的6.5%上调到26.2%。同时该细胞吞噬低剂量的细菌抗原即可有效活化T细胞,其动力学曲线为:1～3h,该细胞抗原递呈能力呈对数增加,3～8hDC抗原递呈能力进入一个平台期,10～12h,抗原递呈能力又迅速增强。结论采用GM-CSF可以从小鼠骨髓中获得大量纯度高并且具有典型免疫表型的DC,该DC细胞能有效的递呈外源性细菌抗原,并具有特殊的抗原递呈动力学曲线。
OBJECTIVE: To study the kinetic characteristics of mouse bone marrow-derived dendritic cells (CSFs) cross-presenting with bacterial particulate antigens. Methods Mouse bone marrow cells were obtained and induced by GM-CSF. The morphology of the cells during the culture was observed with an inverted microscope, and the cell purity and the phenotype were measured by flow cytometry. Day 7 immature DC cells were used for cross-presentation of bacterial antigens and kinetic analysis. Results Mice bone marrow cells were induced by GM-CSF. A large number of dendritic cells with typical morphology and immunophenotype were obtained on the 7th day. The characteristic changes in the phenotypes were also observed after co-stimulation with bacterial lipopolysaccharide The molecular CD80 was increased from 27.7% before stimulation to 71.6%, CD86 from 36.0% before stimulation to 80.3%, and DEC205 from 26.5% before stimulation to 26.2%. While the cells phagocytose low doses of bacterial antigens can effectively activate T cells, the kinetics curve: 1 ~ 3h, the cell antigen presenting capacity increased logarithmically, 3 ~ 8hDC antigen presenting ability into a plateau, 10 ~ 12h, antigen presenting ability and rapidly increased. Conclusion A large number of DCs with high purity and typical immunophenotype can be obtained from mouse bone marrow using GM-CSF. These DCs can effectively deliver exogenous bacterial antigens and have special antigen presenting kinetic curves.