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LymphoPrepTM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6 d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。
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 ofrelated cell surface markers. CD14 ^＋ cells were separated and enriched from human peripheral blood with LymphoPrepTM gradient centrifugation regent and CD14 microbeads. Then the cells were cultured with humanrecombinant granulocyte-macrophage colony-stimulating tactor and human recombinant interleukin 4. The recombinant transforming growth faetor-β1 or β-mercaptoethanol was added into the euhure medium on days 0, 2,4, respectively. On day 6, the cultured cells were harvested and analyzed by flow cytometry. Results showed that the Mo-LC and Mo-IDEC exhibited similar morphological features on day 6. Phenotypical analyses revealed that Mo-IDEC and Mo-LC were all positive to CDla, FcεRI, FcεRII, TLR1, and TLR2. Further, Langerin was expressed solely on the surface of Mo-IDEC, while CD206 on 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.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第3期239-242,共4页
Immunological Journal
基金
国家自然科学基金资助项目(81072447)
