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人DC的获取与DC特异性细胞间黏附分子-3结合非整合素的表达

Generation of DC from human peripheral blood monocytes and expression of DC specific intercellular-adhesion-molecule-3 grabbing nonintegrin
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摘要 目的:证实体外诱导分化的人树突状细胞(DC)表面高效表达树突状细胞特异性细胞间黏附分子-3结合非整合素因子(DC-SIGN),为进一步研究其在丙型肝炎病毒传播中的作用奠定基础。方法:F icoll密度梯度离心法分离外周血单个核细胞(PBMC),黏附2 h后接种于完全培养液中,并加入rhGM-CSF、rhIL-4刺激分化,共培养7天,光镜及扫描电镜观察细胞形态,以DC表面特异性标志DC-SIGN的特异性抗体进行荧光染色,检测DC-SIGN的表达。结果:经诱导产生的细胞具有典型的DC形态特征,高表达DC特异性标志物DC-SIGN。结论:GM-CSF与IL-4联合刺激PBMC可分化为DC,且高表达DC-SIGN,为后续研究DC-SIGN奠定了基础。 Objective ： To determine the expression of DC-SIGN on generated dendritic cells from human peripheral blood. Methods： Peripheral blood monocytes （PBMC） were isolated from blood of healthy individuals by ficoll sedimentation according to standard procedures. Monocytes were purified from PBMC by a 2 h adherence step at 37℃ in complete medium. Nonadherent cells were washed off with PBS and the remaining adherent cells immediately subjected to the DC differentiation protocol. Monocytes resuspended at complete medium containing GM-CSF and IL-4. Cells were cultured for 7 days, with cytokine addition every second day, to obtain a population of immature DC. Characteristics of the cultured cells were observed by LM and SEM, and the expression of DC-SIGN was detected by immunofluorescence （IF）. Results： After 7 days＇culture, typical dendritic cells appeared and the level of DC-SIGNg expression was high. Conclusion ： DC with high expression of DC-SIGN could be induced from peripheral blood monocyte in complete medium containing GM-CSF and IL-4.
作者 李军 冯志华 李光玉 牟丹蕾 王平忠 徐辉
机构地区 第四军医大学唐都医院解放军传染病诊疗中心
出处 《医学研究生学报》 CAS 2005年第12期1066-1068,i0018,共4页 Journal of Medical Postgraduates
基金 国家自然科学基金资助项目(批准号:20170822)
关键词 树突状细胞 树突状细胞特异性细胞间黏附分子-3结合非整合素因子 丙型肝炎病毒 Dendritic cells Dendritic cells specific intercellular-adhesion-molecule-3 grabbing nonintegrin Hepatitis C virus
分类号 R329.2 [医药卫生—人体解剖和组织胚胎学]
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医学研究生学报
2005年 第12期

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