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目的研究不同年龄阶段口腔癌患者外周血单核细胞来源的树突状细胞(dendriticcell,DC)的形态、表型差异并分析其临床意义。方法将2007—2010年兰州大学第一医院及第二医院口腔科住院的口腔鳞癌初诊患者30例,分为40～50岁及≥60岁2组,每组各15例。取患者外周血单核细胞经重组人粒细胞集落刺激因子(rhGM-CSF)、重组人白细胞介素4(rhIL-4)及肿瘤坏死因子-α(TNF-α)诱导培养,获取成熟树突状细胞,光镜观察细胞形态,流式细胞仪检测细胞表面协同刺激分子CD1a、CD83、CD80、CD86的表达。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,其中≥60岁组DC培养过程中细胞逐渐死亡,培养至第7天大部分细胞死亡,获得的DC数量较少,有3例未能培养出DC;而40～50岁组培养细胞的活力保持较好,至第7天均可获得数量较多的DC。结论 2组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,均能扩增出成熟DC,但较40～50岁组,≥60岁组外周血培养的DC体外生长活力明显降低,可能与老年患者免疫功能明显降低有关。
Objective To study the morphological and phenotypic differences of dendritic cells (DCs) derived from peripheral blood mononuclear cells in patients with oral cancer of different ages and to analyze their clinical significance. Methods Thirty new cases of oral squamous cell carcinoma were recruited from Department of Stomatology, First Hospital and Second Hospital of Lanzhou University from 2007 to 2010. The patients were divided into two groups of 40-50 years old and ≥60 years old, 15 cases in each group. Peripheral blood mononuclear cells (PBMCs) were induced by recombinant human granulocyte colony-stimulating factor (rhGM-CSF), recombinant human interleukin-4 (IL-4) and tumor necrosis factor-α (TNF- Cell morphology was observed by light microscope. Flow cytometry was used to detect the expression of costimulatory molecules CD1a, CD83, CD80 and CD86. Results After cultured for 7 days, dendritic cells of typical morphology and phenotype were induced in both groups, of which the cells died gradually in the culture of ≥60 years old. Most of the cells died on the 7th day after culture, and the number of DCs obtained was less , And 3 cases failed to cultivate DC. The viability of cultured cells in 40 ~ 50 years old remained good, and a large number of DCs could be obtained on the 7th day. Conclusions DCs derived from peripheral blood mononuclear cells of two groups of patients with oral squamous cell carcinoma can all grow mature DCs induced by cytokines. However, the growth of DCs cultured in peripheral blood of 40 ~ 50 years old group and ≥ 60 years old group was significantly Reduce, may be related to significantly lower immune function in elderly patients.