腺病毒介导HSP70-HBsAg嵌合基因转染树突状细胞及生物学特性分析

The infection of dendritic cells by recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and Its biological characteristics observations

目的分析携带热休克蛋白70（HSP70）和乙肝表面抗原（HBsAg）嵌合基因的腺病毒表达载体感染人外周血诱导培养树突状细胞（DC）后其生物学特性的改变。方法从正常人外周血中分离单个核细胞，经Ad－HSPT0－HBsAg感染刺激后，在含细胞因子粒细胞巨噬细胞集落刺激因子（GMCSF）、白介素-4（IL4）、肿瘤坏死因子（TNF—α）的培养液体中体外诱导培养DC；采用荧光倒置显微镜、RT-PCR、流式细胞仪（FACS）和混合淋巴细胞反应（MLR）试验对人外周血来源的DC进行生物学特性分析。结果Ad—HSP70－HBsAg感染后的DC能够有效表达示踪蛋白GFP和转录基因HSP70－HBsAg，病毒感染组和对照组的DC在细胞形态、DC细胞表面分子表达和刺激同种异体的人外周血幼稚型T细胞增殖能力的方面无明显差别。结论Ad—HSP70－HBsAg可有效转染DC，感染后的病毒对Dc的生长和生物学特性无显著影响。

Objective To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics. Methods Peripheral blood mononuclear cells （PBMC） were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-α in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer（FACS）, and mixed lymphocyte reaction （MLR）. Results The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70- HBsAg infection. FACS analysis shown that the expression of CDla, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index （SI） between two groups. Conclusion Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.