【摘要】 目的:建立人外周血树突状细胞(DCs)的体外扩增培养技术,并初步探讨其表面CD137、CD137L的表达特点。方法:利用贴壁方法自正常人外周血单个核细胞(PBMC)分离获得单核细胞,体外经重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素4(rhIL-4)联合诱导培养,对培养过程中的细胞进行形态学观察及表型分析,并检测CD137及CD137L的表达。结果:诱导培养5～7天后的非贴壁细胞具有典型的DCs形态,无明显的增殖趋势;流式细胞仪分析显示,MHCⅡ-类分子HLA-DR表达率为64.02%,单核-巨噬细胞系特异性表面标志CD14表达率仅为2.34%;人类DCs表面CD137L表达率为41.03%;CD137表达率为9.77%,LPS刺激后表达率增加为36.06%。结论:人外周血单核细胞,体外经GM-CSF、IL-4诱导,可获得大量的未成熟DCs,其中含有少量的单核-巨噬细胞;此DCs增殖能力很弱,表明单核细胞为DCs的非增殖性前体;体外培养的人类DCs可中等量表达CD137L,并有CD137的表达,LPS刺激可诱导CD137表达的增加。为进一步研究DCs以及CD137、CD137L的生物学特性和功能奠定了基础。更多还原

【Abstract】 Objective:To induce human dendritic cells(DCs)derived from peripheral blood in vitro and to detect the expressions of CD137and CD137L on their surface.Methods:The monocytes were ac-quired from human peripheral blood mononuclear cells(PBMC)by way of adhesion,then cultured with hu-man recombinant granulocyte-macrophage clony stimulating factor and interlukin-4in vitro.The morphology of non-adherent cells was observed by microscope,the related phenotypes of the cells including CD137and CD137L were detected by flow cytometry.Results:After5to7days’ culture,the non-adherent cells had the typical morphology of dendritic cells,which had no significant proliferation trends.Flow cytometry showed these cells expressed64.02%of the MHCⅡclass molecular HLA-DR and2.34%of the monocyte-macrophage lines special marker CD14,at the same time they expressed41.03%of CD137L and9.77%of CD137,when stimulated by LPS they expressed36.06%of CD137.Conclusion:A large amount of imma-ture DCs including very few monocyte-macrophages were acquired from human peripheral blood monocytes cultured with human recombinant granulocyte-macrophage clony stimulating factor and interlukin-4in vitro.These DCs had little ability to proliferate,indicating that the monocyte is the non-proliferating precursor of DCs.These human DCs expressed moderate levels of CD137L and low levels of CD137,and LPS could increase the expression of CD137.All of these will facilitate the further study on the biological characteris-tics and functions of DCs and CD137/CD137L.更多还原