核因子-κB圈套寡核苷酸转染对小鼠成熟树突状细胞生物学特性的影响

Effect of Nuclear Factor Kappa B Decoy Oligodeoxynucleotides Transfection on Biological Characteristics of Mature Dendritic Cells in Mice

目的探讨核因子κB圈套寡核苷酸(NF-κBdecoyODN)转染对小鼠成熟树突状细胞(DCs)生物学特性的影响。方法体外诱导Balb/c小鼠骨髓细胞生成未成熟DCs(imDCs),经抗原OVA以及LPS孵育后成为OVA冲击的骨髓来源成熟DCs(mDCs),流式细胞仪检测DCs表面分子CD11c以及MHC-Ⅱ的表达予以鉴定;将NF-κBdecoyODN体外转染小鼠骨髓来源成熟DCs,同时设立阴性对照组(转染NF-κB"变异"ODN)以及未转染组,EMSA检测转染后NF-κB的活性变化;流式细胞仪检测各组DCs表面共刺激分子(CD40、CD80、CD86)的表达;混合淋巴细胞反应观察各组DCs对OVA致敏的T淋巴细胞增殖的影响。结果成功培养出骨髓来源成熟DCs;NF-κBdecoyODN转染DCs的NF-κB活性明显降低(P<0.05),DCs表面CD40、CD80共刺激分子的表达与阴性对照组、未转染组比较无明显改变(P>0.05),而CD86的表达与其他各组比较明显降低(P<0.05);在混合淋巴细胞反应中,NF-κBdecoyODN转染DCs对T细胞的增殖有抑制作用(P<0.05),而阴性对照组与未转染组的DCs具有强烈的激发T细胞增殖的能力(P<0.05)。结论NF-κBdecoyODN转染DCs可能通过CD86的表达降低显著抑制抗原特异性T细胞活化,该改良的DCs有可能用于哮喘的免疫治疗。

Objective To investigate the effects of nuclear factor kappa B decoy oligodeoxynucleotides （NF-KB decoy ODN） transfection on biological characteristics of mature dendritic cells （mDCs） in mice. Methods hnmature DCs were harvested from Balb/c mice bone marrow,followed by the incubation with antigen OVA and LPS,and mature DCs were evaluated by the expressions of CDllc and MHC- Ⅱ detected by FACS. Mature DCs were transfected with NF-KB decoy ODN and the changes of NF-KB activity after the transfection were detected by EMSA. The expressions of the costimulatory molecules（ CD40, CD80 and CD86 ） on DCs were detected by FACS and the proliferation of T cells was tested by mixed lymphocyte reaction（MLR）. Results The mature DCs were cultured successfully. The NF-κB activity of NF-KB decoy ODN transfected DCs was decreased significantly （ P 〈 0.05 ）. There was no difference in the expressions of CD40 and CD80, but the expression of CD86 was decreased significantly in NF-KB decoy ODN transfection group （ P 〈 0. 05 ）. MLR test showed that the proliferation of T lymphocyte cells was inhibited by NF-KB decoy ODN transfeeted DCs, but was stimulated strongly by the DCs of other groups. Conclusions Mature DCs transfected with NF-κB decoy ODN could inhibit the proliferation and activation of antigenspecical T cells, which was probably related to the down-regulation of CD86 on DCs. This modified DCs might be a promising vaccine for the treatment of asthma in the future.