HPV-11 E7重组腺病毒体外转染树突细胞对其功能的影响

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 MOI 100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。

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

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 aden-ovirus 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-γin the co-culture supematant of DC and T cells.Results The optimal MOI was determined to be 100.After transfection with the optimal MOI of recombinant adenovirus pAD-E7,88% 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.Trans-fected DC still had strong potential to stimulate the proliferation of allogeneic T cells.The levels of cytokines IL-12,IFN-γin 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 signifi-cantly by the transfection with pAD-E7,and transfected DC could induce the polarization of T cells to Th1 cells.