Research of gene transfer into nonhuman promates using novel types of adenovirus vectors

血清型35腺病毒载体是一种很有前途的基因载体。然而，常规小鼠中的Ad35载体的转导概况允许对Ad35载体的转导特性进行有限的估计，这是由于睾丸中亚组B Ad受体的小鼠类似物CD 46的表达受限。为了正确评估Ad35载体的转导特性，我们使用食蟹猴进行了转导实验，食蟹猴以与人类相似的模式普遍表达CD46。体外转导实验表明，培养的食蟹猴细胞有效地转导了Ad35载体。相比之下，Ad35载体介导的转导几乎没有发现在所有的器官，虽然Ad35载体基因组检测到在静脉内给药后的各种器官。与常规Ad血清型5（Ad5）载体注射的猴子相比，在Ad35载体注射的猴子中发现较不严重的组织病理学异常。 ...更多信息 结果表明，结肠炎可引起严重的组织损伤和炎症反应，如肝细胞坏死和结肠淋巴管增生。Ad35和Ad5载体注射后均引起外周血CD3 ^+细胞增加，CD16 ^+细胞和CD20 ^+细胞减少的相似的血液学变化。Ad35载体表现出独特的转导模式，这取决于器官。肝细胞和小胶质细胞分别注射入肝和脑后，主要转导与Ad35载体。肌内注射Ad35载体导致细胞的转导，其呼吁成纤维细胞和巨噬细胞。结膜上皮细胞在输注到眼球玻璃体中后表达转基因表达。在这些器官中，转基因表达仅限于注射点周围的区域。在未注射Ad35载体的器官中未发现转基因表达。这些结果表明，Ad35载体将是一个优选的基因载体，通过直接给药的器官。少

Adenovirus(Ad) serotype 35(Ad35) vectors are promising gene delivery vehicles. However, the transduction profiles of Ad35 vectors in conventional mice allow a limited estimation of transduction properties of Ad35 vectors due to the restricted expression of the mouse analog of the subgroup B Ad receptor, CD46, in the testis. To properly assess the transduction properties of Ad35 vectors, we performed transduction experiments using cynomolgus monkeys, which ubiquitously express CD46 in a pattern similar to that in humans. In vitro transduction experiments demonstrated that cultured cynomolgus monkey cells were efficiently transduced with Ad35 vectors. By contrast, Ad35 vector-mediated transduction was hardly found in the all organs, although Ad35 vector genomes were detected in vanous organs following intravenous administration. Less severe histopathological abnormalities were found in the Ad35 vector-infused monkeys compared with the conventional Ad serotype 5(Ad5) vector-injected monke … More ys, in which serious tissue damages and inflammatory responses, such as hepatocyte necrosis and lymphatic hyperplasia in the colon, were induced. Both Ad35 and Ad5 vectors caused similar hematological changes(increase in CD3^+ cells, and decreases in CD16^+ cells and CD20^+ cells) in peripheral blood cells postinjection.Furthermore, Ad35 vectors were locally injected into the organs. Ad35 vectors exhibited unique transduction patterns depending on the organs. Hepatocytes and microglia were mainly transduced with Ad35 vectors after injection into the liver and brain, respectively. Intramuscular injection of Ad35 vectors resulted in transduction of cells which appealed to be fibroblasts and macrophages. Conjunctival epithelial cells express transgene expression following infusion into the vitreous body of eyeball. In these organs, transgene expression was limited to areas around the injection points. Transgene expression was not found in the organs which did not receive Ad35 vector injection. These results suggest that Ad35 vectors would be a preferable gene delivery vehicle by direct administration in the organs. Less