Ding Zhen-yu, Wang Chun, Su Jing-mei, Wei Yu-quan, Wang Chun-ting
Laboratory of Tumor Biotherapy, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2009 May;40(3):369-73.
To explore the feasibility of constructing a dendritic cell targeted adenovirus as a possible universal tumor vaccine.
The cDNA of human telomerase reverse transcriptase (hTRT) from pBABE-PURO-hTERT was subcloned into pAdTrack-CMV. Recombinant adenovirus for hTRT was constructed in AdEasy system. The confirmed recombinant adenovirus plasmid was transfected into 293 cells to generate the recombinant adenovirus. Later the recombinant adenovirus was multiplied, purified and modified with mannan as a vaccine to immunize mice.
An obvious cytopathic effect was observed with the rescue of recombinant adenovirus. The expression of hTRT was confirmed by PCR and Western blot analysis. Mannan modified adenovirus stained positively by periodic acid Schiff stain, and expression of the adenovirus was found mainly restricted in dendritic cells (DC) in spleen of mice.
A DC-targeted adenovirus was constructed. This might be helpful (or the development of universal tumor vaccine in future.
探索构建树突状细胞靶向腺病毒作为一种可能的通用肿瘤疫苗的可行性。
将来自pBABE-PURO-hTERT的人端粒酶逆转录酶（hTRT）的cDNA亚克隆到pAdTrack-CMV中。在AdEasy系统中构建hTRT的重组腺病毒。将确认的重组腺病毒质粒转染到293细胞中以产生重组腺病毒。随后，将重组腺病毒进行扩增、纯化并用甘露聚糖修饰作为疫苗免疫小鼠。
重组腺病毒拯救成功，观察到明显的细胞病变效应。通过PCR和蛋白质印迹分析确认了hTRT的表达。甘露聚糖修饰的腺病毒经高碘酸希夫染色呈阳性，并且发现腺病毒的表达主要局限于小鼠脾脏中的树突状细胞（DC）。
构建了一种树突状细胞靶向腺病毒。这可能有助于未来通用肿瘤疫苗的开发。
