目的 探討核因子κB 圈套寡核苷酸( NF-κB decoy ODN) 轉(zhuǎn)染對小鼠成熟樹突狀細胞( DCs) 生物學(xué)特性的影響。方法 體外誘導(dǎo)Balb/c 小鼠骨髓細胞生成未成熟DCs( imDCs) , 經(jīng)抗原 OVA 以及LPS孵育后成為OVA 沖擊的骨髓來源成熟DCs( mDCs) , 流式細胞儀檢測DCs 表面分子CD11c 以及MHC-Ⅱ的表達予以鑒定; 將NF-κB decoy ODN 體外轉(zhuǎn)染小鼠骨髓來源成熟DCs, 同時設(shè)立陰性對照組( 轉(zhuǎn)染NF-κB“變異”O(jiān)DN) 以及未轉(zhuǎn)染組, EMSA 檢測轉(zhuǎn)染后NF-κB 的活性變化; 流式細胞儀檢測各組DCs 表面共刺激分子( CD40、CD80、CD86) 的表達; 混合淋巴細胞反應(yīng)觀察各組DCs 對OVA 致敏的T淋巴細胞增殖的影響。結(jié)果 成功培養(yǎng)出骨髓來源成熟DCs; NF-κB decoy ODN 轉(zhuǎn)染DCs 的NF-κB活性明顯降低( P lt;0. 05) , DCs 表面CD40、CD80 共刺激分子的表達與陰性對照組、未轉(zhuǎn)染組比較無明顯改變( P gt; 0. 05) , 而CD86 的表達與其他各組比較明顯降低( P lt;0. 05) ; 在混合淋巴細胞反應(yīng)中,NF-κB decoy ODN轉(zhuǎn)染DCs 對T細胞的增殖有抑制作用( P lt; 0.05) , 而陰性對照組與未轉(zhuǎn)染組的DCs具有強烈的激發(fā)T細胞增殖的能力( P lt; 0. 05) 。結(jié)論 NF-κB decoy ODN 轉(zhuǎn)染DCs 可能通過CD86 的表達降低顯著抑制抗原特異性T細胞活化, 該改良的DCs 有可能用于哮喘的免疫治療。
引用本文： 索立俊,施毅,張方,萬瑛,陳永文,黃鋼,李子玲,宋勇. NF-κB圈套寡核苷酸轉(zhuǎn)染對小鼠成熟樹突狀細胞生物學(xué)特性的影響. 中國呼吸與危重監(jiān)護雜志, 2009, 09(3): 271-274. doi: 復(fù)制
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- 1. Zhou LF, Zhang MS, Hu AH, et al. Selective blockade of NF-kappaB by novel mutated IkappaBalpha suppresses CD3 / CD28-induced activation of memory CD4 + T cells in asthma. Allergy, 2008, 63 :509-517.
- 2. alpha, 25 -dihydroxyvitamin D3 potentiates the beneficial effects of allergen immunotherapy in a mouse model of allergic asthma: role for IL-10 and TGF-beta. J Immunol, 2008, 180: 5211-5221.
- 3. Taher YA, van Esch BC, Hofman GA, et al.
- 4. 孫正, 許哲, 祁小平, 等. 樹突狀細胞-結(jié)直腸癌細胞融合疫苗的制備及生物學(xué)特性的研究. 醫(yī)學(xué)研究生學(xué)報, 2006, 19: 19-22.
- 5. Isomura I, Tsujimura K, Morita A. Antigen-specific peripheral tolerance induced by topical application of NF-kappaB decoy oligodeoxynucleotide. J Invest Dermatol, 2006, 126: 97-104.
- 6. 關(guān)鍵, 鄭碧霞, 程德云. 核因子-κB 在慢性阻塞性肺疾病大鼠肺內(nèi)的表達及潑尼松對其的影響. 中國呼吸與危重監(jiān)護雜志,2008, 7: 52-56.
- 7. Preciado D, Lin J, Wuertz B, et al. Cigarette smoke activates NFkappa B and induces Muc5b expression in mouse middle ear cells.Laryngoscope, 2008, 118: 464-471.
- 8. Wu K, Bi Y, Sun K, et al. Suppression of allergic inflammation by allergen-DNA-modified dendritic cells depends on the induction of Foxp3 + Regulatory T cells. Scand J Immunol, 2008, 67: 140-151.
- 9. Lu L, Lee WC, Takayama T, et al. Genetic engineering of dendritic cells to express immunosuppressive molecules ( viral IL-10, TGFbeta,and CTLA4Ig) . J Leukoc Biol, 1999, 66: 293-296.
- 10. Liu YJ. Thymic stromal lymphopoietin and OX40 ligand pathway in the initiation of dendritic cell-mediated allergic inflammation. J Allergy Clin Immunol, 2007, 120: 238-244.
- 11. Cruz MT, Duarte CB, Gon alo M, et al. LPS induction of I kappa Balpha degradation and iNOS expression in a skin dendritic cell line is prevented by the janus kinase 2 inhibitor, Tyrphostin b42. Nitric Oxide, 2001 , 5: 53-61 .
- 12. Thoeni G, Stoitzner P, Brandacher G, et al. Tetrahydro-4 -aminobiopterin attenuates dendritic cell-induced T cell priming independently from inducible nitric oxide synthase. J Immunol,2005, 174: 7584-7591.