肿瘤坏死因子—α预刺激对培养树突状细胞形态学的影响

目的：研究肿瘤坏死因子（TNF）－α预刺激对树突状细胞形态学的影响，方法：Ficoll密度梯度离心分离健康人外周血，获得单个核细胞，培养基用含有10％胎牛血清的RPMI 1640，分两组进行培养，一组用TNF－α预刺激4h后加入粒细胞－巨噬细胞集落刺激因子（GM－CSF）及白细胞介素－4（IL－4），每48h再次加入上述3种细胞因子；另一组则在培养初始加入GM－CSF及IL－4，每隔48h再次加入上述2种细胞因子，第5天加入TNF－α，两种中所用细胞因子浓度完全一样，且均于第8天收获细胞，前者记作T－DC，后者记作DC。光镜及扫描电镜观察DC形态学变化及差异，结果：T－DC与DC相比，具有更多，更典型的树突状突起，结论：TNF－α预刺激DC前体细胞4h与TNF－α在培养第5天加入相比，所诱导出的DC的树突状形态更为明显，典型。

Effects of tumor necrosis factor-alpha on the morphology of dendritic cells]

OBJECTIVE: To study the effect of tumor necrosis factor-alpha (TNF-alpha) on the morphology and ultrastructure of dendritic cells (DCs). METHODS: Freshly isolated precursors of DCs were cultured in the medium consisting of RPMI 1640 medium and 10 % inactivated fetal bovine serum. In one experiment, the cells were pretreated by TNF-alpha (200 U/ml) for 4 h, followed by treatment with human granulocyte macrophage colony stimulating factor (rhGM-CSF, 800 U/ml) and interleukin-4 (IL-4, 500 U/ml). The above treatments were repeated again every 48 h. The cells in another experiment were treated with rhGM-CSF (800 U/ml) and IL-4 (500 U/ml) on the day of in vitro culture, which was repeated every 48 h with a treatment with TNF-alpha (200 U/ml) on day 5 of cell culture. On day 8, the cells in the 2 parallel experiments were collected with the former cells designated as T-DC and the latter DC. Morphological differences between the 2 group of cells were observed using light microscope and scanning electron microscope. RESULTS: The T-DC exhibited more typical dendric processes than DCs did. CONCLUSION: Pretreatment of the precursors of DCs with TNF-alpha for 4 h, in comparison with TNF-alpha treatment on day 5 of cell culture, produced DCs with more typical and conspicuous dendric morphologies.