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

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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.