人外周血单核细胞源性朗格汉斯细胞_省略_炎症样树突状细胞的诱导及表型分析_邓小蓉

特应性皮炎（atopic dermatitis,AD）是一种慢性复发性炎症性皮肤病，累及约25%儿童及1%～3%成人[1]。朗格汉斯细胞（Langerhans cells，LC）和炎症样树突状表皮细胞（inflammatory dendritic epidermal cells，IDEC）是在AD皮损中出现的2种树突状细胞（dendritic cells，DCs）。其中IDEC是炎症性皮肤病所特有的1种DCs，它仅在炎症性皮肤中表达，被激活后可释放大量炎症因子和趋化因子，不仅在炎症的持续和发展中起着重要作用，还可能与皮损局部免疫反应由Th2反应向Th1反应漂移有关[2-3]。由于这2种细胞在体内数目极少，限制了对其功能的深入研究。陈琛等通过人脐带血体外诱导并成功获得外周血树突状细胞[4]。国外也有研究利用AD患者外周血单核细胞在体外成功诱导了LC样树突状细胞（LC-DC）和IDEC样树突状细胞（IDEC-DC）[5-7]，但目前尚缺乏

·论著·[文章编号]1000-8861（2013）03-0239-04人外周血单核细胞源性朗格汉斯细胞和表皮炎症样树突状细胞的诱导及表型分析

邓小蓉，宋志强，牛军，陈曙光，郝飞*

[摘要]目的利用人外周血单核细胞体外诱导培养朗格罕氏细胞（Mo-LC）和炎症性树突状表皮细胞（Mo-IDEC）并观察其表型特点。方法联合应用LymphoPrep TM梯度离心试剂和CD14+磁珠分离和筛选外周血CD14+单核细胞；在重组人粒细胞-巨噬细胞集落刺激因子（GM-CSF）和重组人白介素4（IL-4）的基础上分别于第0、2、4天加入人天然TGF-β1或β-巯基乙醇（β-ME）。培养第6天时收集细胞，通过相差显微镜观察其形态，并利用单克隆抗体和流式细胞术对诱导细胞亚群检测细胞表面分子的表达水平。结果CD14+单核细胞体外诱导培养6d后可形成具有树突状外观的Mo-LC和Mo-IDEC，2种细胞均不同程度表达CD1a、FcεRI、FcεRII、TLR1、TLR2；Mo-LC表达CD207，Mo-IDEC表达CD206。特应性皮炎患者来源的Mo-IDEC表面CD1a表达高于Mo-LC，且CD23、TLR2、FcεRI的表达水平显著高于健康对照组。结论利用外周血单核细胞可在体外成功诱导Mo-LC和Mo-IDEC，这2种细胞表型特点与体内分离的细胞在形态和表型上类似，为体外进一步研究这2类细胞的功能打下基础。

[关键词]朗格罕氏细胞；表皮炎症性树突状细胞；高亲和力IgE受体；Toll样受体2

[中图分类号]Q786[文献标识码]A

Induction and phenotype analysis of Langerhans cells and inflammatory dendritic epidermal cells derived from human peripheral monocytes

ＤＥＮＧＸｉａｏｒｏｎｇ，ＳＯＮＧＺｈｉｑｉａｎｇ，ＮＩＵＪｕｎ，ＣＨＥＮＳｈｕｇｕａｎｇ，ＨＡＯＦｅｉ*

Department of Dermatology,Southwest Hospital,Third Military Medical University,Chongqing400038,China

*Corresponding Author:HAO Fei,E-mail:haofei62@

[Abstract]This study was designed to establish an in vitro culture system to induce Langerhans cell(Mo-LC) and inflammatory dendritic epidermal cell(Mo-IDEC)from peripheral monocytes,and analyze the expression of related cell surface markers.CD14+cells were separated and enriched from human peripheral blood with LymphoPrep TM gradient centrifugation regent and CD14microbeads.Then the cells were cultured with human recombinant granulocyte-macrophage colony-stimulating factor and human recombinant interleukin 4.The recombinant transforming growth factor-β1orβ-mercaptoethanol was added into the culture medium on days0,2, 4,respectively.On day6,the cultured cells were harvested and analyzed by flow cytometry.Results showed that the Mo-LC and Mo-IDEC exhibited similar morphological features on day6.Phenotypical analyses revealed that Mo-IDEC and Mo-LC were all positive to CD1a,FcεRI,FcεRII,TLR1,and TLR2.Further,Langerin was expressed solely on the surface of Mo-IDEC,while CD206on the Mo-LC.Results in this study indicate that Mo-LC and Mo-IDEC could be generated from human peripheral blood monocytes in vitro and their phenotypical characteristics were similar to that of cells in vivo,thus facilitating further study on the mechanism of inflammatory skin diseases.

[Key words]Langerhans cells;Inflammatory dendritic epidermal cells;High-affinity IgE receptor;Toll-like receptor2

基金项目：国家自然科学基金资助项目（81072447）

作者单位：400038重庆，第三军医大学西南医院皮肤科

*通信作者：郝飞，E-mail：haofei62@