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目的 观察并比较小鼠树突状细胞 (DC)和巨噬细胞对外源性抗原的内吞路径的差异。方法 将小鼠骨髓DC和腹腔巨噬细胞与辣根过氧化物酶 (HRP) 5nm胶体金共孵育 10分钟后再培养 0～ 12 0分钟 ,进行酸性磷酸酶细胞化学反应和组织相容性抗原 (MHC)Ⅱ免疫胶体金标记 ,最后在电镜下观察 5nm胶体金的细胞内运行路径。结果 DC内吞HRP 5nm胶体金 10分钟后再培养 30分钟 ,大部分 5nm胶体金进入MHCⅡ隔室 (MⅡC)中 ,只有极少数酸性磷酸酶阳性的溶酶体内含少量 5nm胶体金。而与DC显著不同的是巨噬细胞摄入的大部分 5nm胶体金进入了溶酶体 ,还有部分 5nm胶体金进入MⅡC中。内吞后再培养 60分钟 ,DC内仍有许多 5nm胶体金 ,而巨噬细胞内已很少见到 5nm胶体金。结论 同一种外源性抗原被DC和巨噬细胞内吞后 ,在这两类细胞中的运行路径明显不同。内吞后 30分钟 ,巨噬细胞摄入的大部分抗原进入溶酶体 ,而DC则将大部分抗原送入MⅡC ,这可能与DC独特的抗原提呈功能有关。另外 ,巨噬细胞对抗原的清除能力比DC强得多。
Objective To observe and compare the difference of endonuclease pathways of exogenous antigens between dendritic cells (DCs) and macrophages in mice. METHODS: Mouse bone marrow DCs and peritoneal macrophages were co-incubated with horseradish peroxidase (HRP) 5nm colloidal gold for 10 minutes and then cultured for 0-1200 minutes for acid phosphatase cytochemical reaction and histocompatibility antigen (MHC) Ⅱ immunogold labeling. Finally, the intracellular path of 5nm colloidal gold was observed under electron microscope. Results DC endocytosed HRP 5nm colloidal gold 10 minutes and then cultured for 30 minutes, most of the 5nm colloidal gold into the MHC Ⅱ compartment (M Ⅱ C), only a very small number of acid phosphatase-positive lysosomes contain a small amount of 5nm colloidal gold. The significant difference from DC is that most of the 5 nm colloidal gold taken up by macrophages enters the lysosome, and some 5 nm colloidal gold enters the M II C. After the endocytosis and then cultured for 60 minutes, there are still many 5nm colloidal gold in DC, while macrophages have rarely seen 5nm colloidal gold. Conclusion The same exogenous antigen was endocytosed by DCs and macrophages, and the pathways of operation in these two types of cells were significantly different. Thirty minutes after endocytosis, most of the macrophages ingested the antigen into the lysosome, while DC sent most of the antigen into MIIC, which may be related to the unique antigen presenting function of DC. In addition, macrophages are much less able to clear antigens than DC.