NF-κB圈套寡核苷酸轉染對小鼠成熟樹突狀細胞生物學特性的影響_《中國呼吸與危重監護雜志》

作者：

索立俊 ¹ ,  施毅 ¹ , 張方 ¹ , 萬瑛 ² , 陳永文 ² , 黃鋼 ³ , 李子玲 ¹ , 宋勇 ¹

1.南京大學醫學院臨床學院（南京軍區南京總醫院）呼吸內科（江蘇南京 210002）2.第三軍醫大學免疫研究所;;
3.第三軍醫大學遺傳學教研室;

關鍵詞：

核因子-κB 圈套寡核苷酸樹突狀細胞免疫耐受哮喘

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目的探討核因子κB 圈套寡核苷酸( NF-κB decoy ODN) 轉染對小鼠成熟樹突狀細胞( DCs) 生物學特性的影響。方法體外誘導Balb/c 小鼠骨髓細胞生成未成熟DCs( imDCs) , 經抗原 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 體外轉染小鼠骨髓來源成熟DCs, 同時設立陰性對照組( 轉染NF-κB“變異”ODN) 以及未轉染組, EMSA 檢測轉染后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; 混合淋巴細胞反應觀察各組DCs 對OVA 致敏的T淋巴細胞增殖的影響。結果成功培養出骨髓來源成熟DCs; NF-κB decoy ODN 轉染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中,NF-κB decoy ODN轉染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染組的DCs具有強烈的激發T細胞增殖的能力( P lt; 0. 05) 。結論 NF-κB decoy ODN 轉染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圈套寡核苷酸轉染對小鼠成熟樹突狀細胞生物學特性的影響. 中國呼吸與危重監護雜志, 2009, 09(3): 271-274. doi: 復制

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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.

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-結直腸癌細胞融合疫苗的制備及生物學特性的研究. 醫學研究生學報, 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. 關鍵, 鄭碧霞, 程德云. 核因子-κB 在慢性阻塞性肺疾病大鼠肺內的表達及潑尼松對其的影響. 中國呼吸與危重監護雜志,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.

《中國呼吸與危重監護雜志》

NF-κB圈套寡核苷酸轉染對小鼠成熟樹突狀細胞生物學特性的影響

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《中國呼吸與危重監護雜志》

NF-κB圈套寡核苷酸轉染對小鼠成熟樹突狀細胞生物學特性的影響

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