口腔癌患者外周血树突状细胞的生物学特性

Biological characteristics of monocyte-derived dendritic cells from peripheral blood of patients with oral carcinoma

目的:研究口腔癌患者外周血树突状细胞(DC)的形态、表型和功能性变化及其临床意义.方法:口腔癌患者(实验组)及正常人(对照组)15例外周血单个核细胞经GM-CSF,IL-4及TNF-α诱导培养,获取成熟DC,光镜和电镜观察细胞的形态,流式细胞仪检测细胞表型如CD1a,CD83,CD80,CD86和HLA-DR等分子的表达变化,用噻唑蓝(MTT)法检测DC与自体和同种异体的混合淋巴细胞反应(MLR)中对淋巴细胞增殖的刺激能力.结果:正常人及口腔癌患者的外周血单个核细胞经体外7d诱导培养,均诱导出具有典型形态和表型的DC,其中实验组细胞的CD83,CD1a表达率分别为49.3±9.5和48.1±7.3,与对照组两者表达率62.1±7.9和60.2±6.8相比,差异有统计学意义(P<0.05),CD86,CD80和HLA-DR的表达率分别为85.5±8.7,83.2±8.1和78.3±5.4,与对照组三者表达率89.2±6.7,87.7±7.2和82.3±4.1相比无明显统计学差异(P>0.05);实验组DC细胞CD83,CD1a,CD86,CD80和HLA-DR表达的平均荧光指数(MFI)分别为4.80±2.13,3.96±1.15,12.33±6.75,19.63±8.12和39.44±7.9,均低于对照组,但差异无统计学意义(P>0.05);口腔癌患者来源的DC在体外具有诱导同种异体和自体外周血T细胞增殖的能力.结论:口腔癌患者外周血单核细胞来源可诱导成为具有功能性的成熟DC,该细胞可应用于口腔癌的免疫治疗.

AIM： To investigate the morphology, phenotype and functions of dendritic cell （DC） from peripheral blood of the healthy individuals and the patients with oral carcinoma. METHODS： Fifteen patients with oral carcinoma at stages of II or above were assigned to study group and 15 healthy volunteers to control group. Monocytes were isolated by density centrifugation, and cultured in the presence of GM-CSF, IL-4 and TNF-α for 7 d in vitro. The morphological change of the cultured cells was observed by hght microscopy and electron microscopy. The expressions of CD83, CD1a, CD86, CD80, HLA-DR were analyzed by flow cytometry. The ability of DC to stimulate the proliferation of lymphocyte in mixed lymphocyte reactions （MLR） was determined with the method of MTT. RESULTS： Morphologically and phenotypically typical DC was obtained in the 2 groups after culture for 7 d. The expression rates of CD83, CD1 a on DC from the patients were 49.3 ± 9.5 and 48.1 ±7.3 respectively, significantly lower than those from healthy volunteers （ P 〈 0.05 ）. The DC from the patients had the lower expression levels of CD86, CD80 and HLA-DR （85.5 ±8.7, 83.2 ±8.1 and 78.3 ±5.4） than healthy volunteers did （ 89.2 ± 6.7, 87.7 ± 7.2 and 82.3 ± 4.1）, but not significantly （ P 〉 0.05 ）. Moreover, the mean fluorescence intensity of CD83, CD1a, CD80, CD86 and HLA- DR expressed in study group were 4. 80 ± 2. 13, 3.96 ± 1.15, 19.63 ±8. 12, 12.33 ±6.75 and 39.44 ±7.9, respectively, which were lower but not statistically different from the control group （P 〉 0.05 ）. In addition, DC from the study group had the ability to stimulate the proliferation of allogeneic and autogenous lymphocytes in vitro. CONCLUSION： Monocytes in peripheral blood of the patients with oral carcinoma can be induced to differentiate into mature DC with the ability to stimulate the proliferation of lymphocytes, and so these DC might be used in immunotherapy for oral carcinoma.