单核细胞源性树突状细胞表达淋巴细胞趋化因子mRNA的初步研究  

Preliminary study on lymphotactin expression of monocyte-derived dendritic cells

作　　者：鲁雄兵[1] 谢辉莉[1] 黄红卫[1] 史子敏[1] 熊建华[1] 潘正跃[1] 

机构地区：[1]南昌大学第二附属医院泌尿外科,江西南昌330006

出　　处：《细胞与分子免疫学杂志》2006年第6期723-725,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30271300)

摘　　要：目的:体外诱导、培养单核细胞源性树突状细胞(DC),研究其淋巴细胞趋化因子(lymphotactin,Lptn)mRNA表达的动态变化。方法:采用密度梯度离心的方法分离人外周血中的单个核细胞(PBMC),用重组人粒细胞-巨噬细胞集落群体刺激因子(rhGM-CSF)、重组人白细胞介素-4(rhIL-4)刺激贴壁的单核细胞,诱导培养DC,第6天用重组人肿瘤坏死因子-α(rhTNF-α)诱导DC成熟。用流式细胞术检测成熟和未成熟的DC表面分子CD1a和CD83;在电镜下观察成熟DC的形态;以RT-PCR法扩增其LptncDNA并克隆至pGM-TEasyT载体中,测序;以RT-PCR结合凝胶成像分析系统,半定量分析培养3、5及7dDC的LptnmRNA表达的强度。结果:电镜观察培养7d的细胞具有典型的DC形态,流式细胞术检测DC表面分子CD83呈高水平表达。用RT-PCR法克隆的cDNA序列与GenBank中U23772(登陆号)提供的序列一致。培养3d的DC不表达LptnmRNA,培养7d的DC较培养5d的DCLptnmRNA表达增强。结论:单核细胞源性DC能表达LptnmRNA,随着DC的成熟,LptnmRNA的表达增强。AIM ： To induce dendritic cells （DCs） from human peripheral blood monocytes in vitro and to investigate the kinetics lymphotactin （Lptn） mRNA expression. METHODS： Monocytes were isolated from normal human peripheral blood cells by density gradient centrifugation and the plastic-adherent cells were cultured with the cytokines （rhGM-CSF, rhIL-4, rhTNF-α）. The immature and mature DCs＇ surface antigens CDIa and CD83 were analyzed by flow cytometry （FCM）. The morphology of mature DCs induced by rhTNF-α was observed under electron microscope. Lptn cDNA was amplified by RT-PCR, cloned to vector pGM-T Easy and completely sequenced. The RT-PCR products of cells were separated on gel and analyzed by semi-quantification. RESULTS： The cells cultured for 7 d exhibited special dendritic morphology of mature DCs and highly expressed CD83. The sequence of cDNA was confirmed and was identical to that of NO. U23772 published in GenBank. The Lptn mRNA expression of DCs was stronger in DCs cultured for 7 d than in those for 5 d, whereas it was not detected in those for 3 d. CONCLUSION： The monocyte-derived dendritic cells can express Lptn mRNA in a maturation-dependent manner.

关 键 词：树突状细胞 淋巴细胞趋化因子 单核细胞 

分 类 号：R392.12[医药卫生—免疫学]