Smahel M, Sobotkova E, Vonka V, Hamsikova E, Zak R, Kitasato H, Ludvikova V
Department of Experimental Virology, Institute of Hematology and Blood Transfusion, 128 20 Prague 2, Czech Republic.
Oncol Rep. 1999 Jan-Feb;6(1):211-5. doi: 10.3892/or.6.1.211.
The capability of DNA to elicit anti-tumour immunity was studied using human papillomavirus type 16 (HPV16)-transformed Syrian hamster cells denoted K3/II. These cells had been derived after cotransfection of primary kidney cell cultures with p16HHMo plasmid containing E6/E7 oncogenes of HPV16 and pEJ6.6 plasmid containing the activated human H-ras oncogene; they express both the HPV16 and activated H-ras genes. As a DNA vaccine, the p16HHMo plasmid was used. Three doses of the plasmid (either 100 microg or 10-15 microg per dose) were administered intramuscularly at 3-week intervals. The animals were challenged with four different doses (10(3)-10(6) per animal) of K3/II cells 10 days after the last plasmid injection. In one experiment the lower dose of plasmid DNA was also given in a mixture with the cationic lipid DOTAP. In another experiment, the pEJ6.6 plasmid (100 microg per dose) was used either alone or in combination with p16HHMo. In all experiments animals inoculated with the same doses of pBR322 plasmid served as controls. A moderate protective effect was observed in animals inoculated with the 100-microg doses of p16HHMo, but not in those inoculated with 10-15 microg of the same plasmid, whether given with or without DOTAP. A protective effect was also observed after administration of the pEJ6. 6 plasmid. At the time of challenge a portion of the p16HHMo-immunized, but not the pBR322-treated, animals possessed antibodies reactive in ELISA with peptides derived from the N-terminal portion of HPV16 E7 protein and with one peptide derived from E6 protein, while two other E6 peptides exhibited non-specific reactivity.
利用人乳头瘤病毒16型（HPV16）转化的叙利亚仓鼠细胞（命名为K3/II）研究了DNA引发抗肿瘤免疫的能力。这些细胞是在原代肾细胞培养物与含有HPV16 E6/E7癌基因的p16HHMo质粒和含有激活的人H-ras癌基因的pEJ6.6质粒共转染后获得的；它们同时表达HPV16和激活的H-ras基因。作为DNA疫苗，使用了p16HHMo质粒。以3周的间隔肌肉注射三剂该质粒（每剂100微克或10 - 15微克）。在最后一次质粒注射10天后，用四种不同剂量（每只动物10³ - 10⁶个）的K3/II细胞对动物进行攻击。在一项实验中，较低剂量的质粒DNA也与阳离子脂质DOTAP混合给药。在另一项实验中，单独或与p16HHMo联合使用pEJ6.6质粒（每剂100微克）。在所有实验中，接种相同剂量pBR322质粒的动物作为对照。在接种100微克剂量p16HHMo的动物中观察到中等程度的保护作用，但在接种相同质粒10 - 15微克的动物中未观察到，无论是否与DOTAP一起给药。在施用pEJ6.6质粒后也观察到了保护作用。在攻击时，一部分接种p16HHMo的动物（而非接种pBR322处理的动物）在ELISA中具有与源自HPV16 E7蛋白N端部分的肽以及一种源自E6蛋白的肽发生反应的抗体，而另外两种E6肽表现出非特异性反应。
