Enhancement of anti-cancer effect with genetically modified adenovirus in gene therapy

根据研究计划，在重组腺病毒5/3和5/3嵌合腺病毒的基础上，利用人VEGF启动子构建了连续复制型腺病毒Ad 5VEGFE 1和Ad 5/3VEGFE 1。接着，通过将Ad 5VEGFE 1和Ad 5/3VEGFE 1共感染到HEK 293细胞中，制备在同一病毒体上具有两种血清型腺病毒纤维的嵌合腺病毒。血清5型与血清5/3型的混合感染比例为7：3时，嵌合腺病毒的感染率最高。大量病毒制备后，按既定规律在CsCl_2溶液中进行超离心浓缩。然后通过透析精制病毒溶液。收集的病毒溶液的密度的产率在10^9至10^pfu/mL中是比较优异<10>的。2004财年报告中报告了体外实验结果。三种癌细胞系C33 A、NCI-H157和SKOV用于体内实验。这些细胞系已经检查了它们在裸鼠皮下空间中的肿瘤形成能力。在这些细胞系形成肿瘤后，分别向每个肿瘤注射10^9 pfu的Ad 5VEGFE 1、Ad 5/3VEGFE 1和AdmsVEGFE 1。治疗后测量肿瘤直径以评估肿瘤增殖。结果表明，Ad 5VEGFE 1和AdmsVEGFE 1对C33 A和NCI-H157肿瘤的抗肿瘤作用以及体外实验结果均较高。此外，Ad 5/3VEGFE 1和AdmsVEGFE 1对SKOV肿瘤的生长抑制效果极佳。而AdmsVEGFE 1的作用位于Ad 5VEGFE 1和Ad 5/3VEGFE 1作用的中间。据认为，在体内肿瘤中没有发生足够的花叶病毒产生。基因修饰的腺病毒被认为是有用的，以加强在复制型腺病毒的抗肿瘤作用。

Conditionally replicative Adenoviruses, Ad5VEGFE1 and Ad5/3VEGFE1, were made using human VEGF promoter based on serotyp 5 adenovirus and 5/3 chimeric adenovirus according to the research plan. Next, mosaic adnovirus possessing both serotype adenovirus fibers on the same virion was made from co-infection of Ad5VEGFE1 and Ad5/3VEGFE1 into the HEK293 cell. Co-infection ratio of serotype 5 and serotype 5/3 of 7:3 showed the highest value to make a mosaic adenovirus. It concentrated by the ultracentrifugation in the CsCl_2 solution according to the established rule after a large amount of viral preparation. The virus solution was refined by the dialysis afterwards. Yield of the density of the collected virus solution was comparatively excellent in 10^9 to 10^<10> pfu/mL. In vitro experimet results was reported in 2004 fiscal year report. Three cancer cell lines, C33A, NCI-H157, and SKOV were used for the in vivo experiment. These cell lines already checked for their tumor formation ability in the subcutaneous space of the nude mouse. After tumor formation of these cell lines, each tumor was injected by 10^9 pfu of Ad5VEGFE1, Ad5/3VEGFE1, and AdmsVEGFE1 respectively. The tumor diameter was measured to evaluate the tumor proliferation after treatment. As a result, the antitumor effects of Ad5VEGFE1 and AdmsVEGFE1 are higher for C33A and NCI-H157 tumors as well as the in vitro experiment result. Also, the growth suppressive effect of Ad5/3VEGFE1 and AdmsVEGFE1 was excellent for the SKOV tumor. However, the effect of AdmsVEGFE1 was located in the middle in the effect of Ad5VEGFE1 and Ad5/3VEGFE1. It was thought that an enough mosaic virus generation did not happen in the tumor in vivo. Genetically modified adenovirus was thought to be useful for the reinforcement of the antitumor effect in the replicative adenovirus.