HPV-11 E7重组腺病毒体外转染树突细胞对其功能的影响  

In vitro effect of transfection with recombinant adenovirus encoding human papillomavirus type 11(HPV-11)E7 on dendritic cells

作　　者：王飞[1] 相文忠[2] 毕志刚[2] 李光富[3] 刘丰[3] 吴海玮[3] 张兆松[3] 

机构地区：[1]东南大学附属中大医院皮肤科,南京210009 [2]南京医科大学第一附属医院皮肤科 [3]南京医科大学分子生物研究所

出　　处：《中华皮肤科杂志》2007年第5期289-292,共4页Chinese Journal of Dermatology

基　　金：江苏省重点学科基金(135-3);东南大学中大医院科研基金(2005YJ04)

摘　　要：目的研究腺病毒载体介导HPV-11 E7基因转染树突细胞（DC）对其表型和功能的影响。方法用最佳感染滴度（MOI）重组腺病毒pAD—E7转染成熟DC，流式细胞仪检测转染前后DC表型变化,并用^3H—TdR掺入法检测转染前后DC刺激同种淋巴细胞增殖的能力及ELISA检测DC、T细胞共同培养上清中细胞因子的水平。结果MOI100为pAD—E7转染DC最佳滴度，pAD—E7转染成熟DC前后对细胞表面的特征性表型CD1a、CD83及CD40、HLA—DR无影响，转染后的DC仍具有较强的刺激同种异体淋巴细胞增殖的能力，能刺激T细胞分泌大量IFN-γ、IL-12。结论pAD—E7转染成熟DC对其功能无明显影响．转染后的DC与T细胞共孵育能诱导T细胞向Th1细胞极化。Objective To investigate the in vitro effect of transfection with recombinant adenovirus encoding HPV-11 E7 gene on the phenotype and function of dendritic cells （ DC ）. Methods Optimal multiplicity of infection （MOI） was determined. Mature DC were transfected with the recombinant adenovirus encoding HPV-11 E7 gene （ namely pAD-E7 ） at the optimal MOI. The expression of E7 protein in mature DC and of surface markers （CD1a, CD83, CD40, CD80 and HLA-DR ） on these cells were assessed by flow cytometry before and after the transfection. Meanwhile, the function of stimulating the proliferation of allogeneic T cells by DC was detected by 3H-thymidine incorporation method. ELISA was used to detect the level of cytokines IL-10, IL-12, IFN-γ in the co-culture supernatant of DC and T cells. Results The optimal MOI was determined to be 100. After transfection with the optimal MOI of recombinant adenovirus pAD-ET, 88% mature DC expressed E7 protein, and more than 93% of these cells were viable. No significant changes in surface markers were examined on transfected DC from those without transfection. Transfected DC still had strong potential to stimulate the proliferation of allogeneic T cells. The levels of cytokines IL-12, IFN-γ in co-culture supernatant were increased significantly after the transfection, but no significant change was observed for the level of IL-10. Conclusions The function of mature DC is not affected significantly by the transfection with pAD-ET, and transfected DC could induce the polarization of T cells to Thl cells.

关 键 词：尖锐湿疣 遗传载体 乳头状瘤病毒 人 树突细胞 

分 类 号：R752.53[医药卫生—皮肤病学与性病学]