靶向防龋质粒转染的树突状细胞抗原提呈相关基因表达研究

Gene Expression Profiles of Targeted Anti-caries DNA Plasmid Transfected Dendritic Cells Using Gene Arrays.

目的:比较靶向和非靶向防龋质粒转染的树突状细胞抗原提呈相关基因表达差异,分析靶向防龋质粒增强抗体反应的可能机制。方法:构建绿色荧光蛋白标记的靶向防龋质粒pMGJGLU/GFP和非靶向防龋质粒pCDG-LU/GFP。pMGJGLU/GFP和pCDGLU/GFP分别转染小鼠树突状细胞,芯片分析两组树突状细胞抗原提呈相关基因的表达差异,实时荧光定量RT-PCR检验差异表达基因。结果:与pCDGLU/GFP转染的树突状细胞相比,pMGJGLU/GFP转染的树突状细胞有28个基因表达上调,6个基因表达下调,其中Ccl17、Ccl19和CD209a上调最为显著。结论:靶向DNA质粒转染树突状细胞一系列活化和成熟基因发生了变化,其中Ccl17、Ccl19和CD209a可能发挥重要的免疫增强作用。

Objective： To analysis the gene expression profiles of the targeted anti - caries DNA plasmid transfected dendritic cells （DCs） and explore the possible mechanism underlying the enhanced immune responses after targeted anti - caries DNA plasmid vaccination. Methods： A green fluorescent protein （GFP） labeled targeted anti - caries plasmid pMGJGLU/GFP and a GFP labeled non - targeted anti - caries plasmid pCDGLU/GFP were constructed. The expression profiles of 176 immunologieally relevant genes of pMGJGLU/GFP and pCDGLU/GFP transfeeted DCs were investigated by gene array analysis. Real - time RT - PCR analysis was used to verify the up - regulation of the mRNA expression levels of selected genes. Results： Overall, the expression levels of 28 genes were up - regulated in pMGJGLU/GFP transfected DCs compared with pCDGLU/GFP transfected DCs. Notably, we identified the specific up - regulation of mRNA expression levels of cytokines Ccll7, Ccll9, and antigen presentation receptor Cd209a of pMGJGLU/GFP transfected DCs. Conclusion： Numerous genes related to the activation and maturation of DCs were obviously changed. Cc117, Cc119 and Cd209a may play an important role in the enhanced immune responses.