口腔癌患者外周血树突状细胞的体外培养及表型分析

In vitro culture and phenotype analysis of monocyte-derived dendritic cell from peripheral blood of patients with oral carcinoma

目的:研究口腔癌患者外周血单核细胞来源的树突状细胞(dendritic cell,DC)形态、表型,与正常人进行比较并分析其临床意义。方法:15例口腔癌患者(实验组)及正常人(对照组)外周血单核细胞经GM-CSF,IL-4及TNF-α诱导培养,获取成熟树突状细胞,光镜和电镜观察细胞形态,流式细胞仪检测细胞表型CD1a、CD83、CD80、CD86和HLA-DR的表达变化。结果:培养7d后,2组均诱导出典型形态和表型的树突状细胞,实验组CD83、CD1a的表达率较对照组低,差异有统计学意义(均P<0.001),但CD86、CD80、HLA-DR表达率2组差异无统计学意义(均P>0.05)。结论:口腔鳞癌患者外周血单核细胞来源的DC经细胞因子诱导培养,能扩增出较大量高纯度并保持功能活性的成熟DC,为进一步开展以DC为基础的生物治疗和研究打下了基础。

Objective： To compare the phenotype of induced dendritic cell （DC） derived from peripheral blood of the healthy individuals and the patients with oral carcinoma. Methods： 15 patients with oral carcinomal at stages II or above were assigned to study group and 15 cases of healthy volunteers were assigned to control group. Leukocyte were isolated by density centrifugation. Adherent fractions were incubated in present of GM-CSF, IL-4 and TNF-α for 7 days in vitro. The morphological change of the cultured cells was observed by light microscopy and electronic microscopy. The phenotype expressions of CD83, CD1 a, CD86, CD80, HLA-DR were analysized by flow eytometry. Results：Typical DCs were obtained from two groups after 7 days culture. The expression rate of CD83, CD1 a on DC from healthy volunteers was significantly higher than that from patients （ P 〈 0. 001 ）. The DC from healthy volunteers had the higher expression level of CD80, CD86 and HLA-DR than that from patients, but with no statistically difference （P 〉 0. 05 ）. Condusion： DC derived from oral cancer patient＇s peripheral blood monocyte might be more matured and is a good cell model for further inununotherapy and research.