NF-κB圈套性寡核苷酸诱导骨髓DC疫苗的实验研究

Reseach in Bone Marrow-derived DCs Modified with NF-κB Decoy Oligodeoxynucleotides

目的体外诱导、培养大鼠(Lewis大鼠)骨髓来源改良树突状细胞(DC),为进一步研究核苷酸圈套技术抗移植排斥反应作准备。方法Lewis大鼠骨髓造血干细胞,体外经重组大鼠细胞因子GM-CSF、IL-4诱导成DC;核因子-κB(NF-κB)圈套寡核苷酸(NF-κB decoy ODN)用于阻止DC的成熟,降低细胞表面分子的表达,制备改良的DC;LPS用于刺激DC的成熟。流式细胞仪检测DC表面分子CD11c、CD80、CD86等的表达情况,分析DC的细胞特性。结果体外GM-CSF和IL-4诱导的大鼠骨髓来源DC纯度高、数量丰富。NF-κB decoy ODN能明显降低DC表面分子(CD11c、CD80、CD86等)的表达,表现为非成熟状态;经LPS刺激仍能保持非成熟状态。结论经NF-κB decoy ODN处理的大鼠骨髓来源DC,其状态稳定,低表达表面分子和共刺激分子,可作为进一步研究的DC疫苗。

Objective To obtain Lewis rats bone marrow-derived modified DCs by using NF-kB decoy oligodeoxynucleotides （INF-kB decoy ODN）. Methods DCs were propagated from Lewis rats bone marrow with GM-CSF and IL-4; their maturation was arrested by treatment with ODN specifically against nuclear factor NF-kB; and the modified DCs were stimulated by LPS. DC phenotype （CDllc, CDSO, CD86） was examined by flow cytometry. Results Compared with mature DCs, NF-kB decoy ODN-treated DCs exhibited features of immature DCs, with low level of surface molecule and costimulatory molecule （CD11c, CDSO, CD86） expression. After being stimulated by LPS, the modified DCs （NF-kB decoy ODN-treated DCs） still had the features of immature DCs. Conclusion The modified DCs, with NF-kB decoy ODN-treated, exhibit features of immature DCs steadly; and can be used as a ＂vaccine＂.