肿瘤相关巨噬细胞与HUT-78淋巴瘤细胞相互作用的实验研究  

Interaction between Tumor-associated Macrophages( TAMs) and HUT-78 Lymphoma Cells

作　　者：魏冲[1] 王昊天[1] 姜志超[1] 张薇[1] 曹欣欣[1] 李天骄[1] 汪玄[1] 周道斌[1] 

机构地区：[1]中国医学科学院、北京协和医学院北京协和医院血液内科,北京100730

出　　处：《中国实验血液学杂志》2014年第4期988-994,共7页Journal of Experimental Hematology

基　　金：北京市自然科学基金(7122140)

摘　　要：本研究旨在探究淋巴瘤细胞对单核细胞向肿瘤相关巨噬细胞(TAM)分化的影响以及TAM对淋巴瘤细胞生长的作用。通过密度梯度离心法及免疫磁珠分选获得初治淋巴瘤患者及健康志愿者外周血单核细胞,利用Transwell嵌套建立单核细胞与淋巴瘤细胞株HUT-78的共培养模型。通过流式细胞术分析TAM细胞CD68、CD163的表达,应用细胞计数法绘制淋巴瘤细胞的生长曲线,ELISA法检测共培养体系中细胞因子IL-10、VEGF的含量。结果表明,单核细胞与淋巴瘤细胞共培养后CD68、CD163、CD68&CD163(+)细胞的比例显著升高(P<0.05),在患者及健康志愿者升高程度未见显著差异,且二者的单核细胞及其分化而来的巨噬细胞在体外培养条件下对淋巴瘤细胞的生长无直接的促进或抑制作用。患者细胞共培养组IL-10、VEGF水平明显低于单核细胞和淋巴瘤细胞单独培养组之和(P<0.05),而在健康志愿者无此现象。结论:淋巴瘤细胞能促进外周血单核细胞向M2型巨噬细胞分化,在作用程度上,患者及健康志愿者未见显著差异。由患者单核细胞转化而来的TAM在共培养体系中的作用类似M1型巨噬细胞,能有效抑制共培养体系中总IL-10和VEGF的分泌。This study was aimed to explore the effects of lymphoma cells on the differentiation of monocytes from peripheral blood to tumor-associated macrophages （TAM） and the effect of TAM on proliferation of lymphoma cells in vitro, and investigate the difference between newly diagnosed lymphoma patients and healthy volunteers. Blood samples were obtained from 15 newly diagnosed lymphoma patients and 8 healthy volunteers. Monocytes from peripheral blood were isolated by Ficoll- Hypaque density gradient centrifugation and CD14 immuno-magnetic beads. Then monocytes were directly co-cultured with HUT-78 lymphoma ceils by using TransweU apparatus in vitro. Expression of the markers of TAM （CD68 and CD163） were detected by flow cytometry to analyse the proportion of differentiated TAM. Growth curve of HUT-78 ceils was made by direct cell count. The IL-10 and VEGF levels in the co-culture system were detected by ELISA. The detection results of newly diagnosed lymphoma patients were compared with that of healthy controls. The results showed that the proportion of CD68（ ＋ ） ,CD163（ ＋ ） and CD68 ＋ CD163（ ＋ ）cells were significantly up- regulated after co-cultured with HUT-78 lymphoma cells in both patients and healthy controls （ P 〈 0, 05 ）. There was no statistical significance in the increasing degree between patients and healthy controls. TAM differentiated from peripheral blood monocytes showed no significant promotion or inhibition on the growth of co-cultured lymphoma cells. For patients, the IL-10 and VEGF levels in the co-culture group were significantly lower than those in two single culture groups （P 〈 0.05）. For healthy controls, there was no significant difference between these two. It is concluded that lymphoma cells can promote the differentiation of monocytes to macrophages with M2-1ike phenotype. There is no difference in the promoting degree between patients and healthy controls. TAM differentiated from patients＇monocytes significantly down- regulate levels of 1L-10 and V

关 键 词：淋巴瘤 肿瘤相关巨噬细胞 单核细胞 

分 类 号：R733.1[医药卫生—肿瘤]