McClane S J, Hamilton T E, DeMatteo R P, Burke C, Raper S E
Harrison Department of Surgical Research, University of Pennsylvania School of Medicine, Philadelphia, USA.
Pancreas. 1997 Oct;15(3):236-45. doi: 10.1097/00006676-199710000-00004.
Gene transfer technology may provide a novel approach to treatment for pancreatic diseases. Recombinant adenovirus achieves efficient gene transfer in vivo. In this study, a murine model of adenoviral-mediated pancreatic gene transfer was developed, and the factors responsible for adenoviral elimination were investigated. Three days after direct pancreatic injection of a replication-defective adenovirus containing the lacZ transgene, a high proportion (76.8 +/- 6.7%) of pancreatic cells expressed beta-galactosidase, the gene product. Gene expression was absent by 28 days posttransduction. In immunodeficient mice, beta-galactosidase expression persisted with 20.0 +/- 6.0% of pancreatic cells staining positive 60 days after viral transduction. To test whether early viral proteins are the antigenic components responsible for the potent antiviral immune response, normal mice were injected with different adenoviral vectors containing early gene deletions. Vectors containing deletions in early region 2 or 4 expressed beta-galactosidase at 28 days. Presently available adenoviral vectors engineered to avoid this response offer minimal improvements in transgene duration. Further vector modifications or alternative strategies are needed to achieve stable pancreatic adenoviral transgene expression.
基因转移技术可能为胰腺疾病的治疗提供一种新方法。重组腺病毒可在体内实现高效的基因转移。在本研究中，建立了腺病毒介导的胰腺基因转移小鼠模型，并对腺病毒清除的相关因素进行了研究。在胰腺直接注射含lacZ转基因的复制缺陷型腺病毒三天后，高比例（76.8±6.7%）的胰腺细胞表达了基因产物β-半乳糖苷酶。转导后28天基因表达消失。在免疫缺陷小鼠中，病毒转导60天后，β-半乳糖苷酶表达持续存在，20.0±6.0%的胰腺细胞染色呈阳性。为了测试早期病毒蛋白是否是引发强效抗病毒免疫反应的抗原成分，给正常小鼠注射了含有早期基因缺失的不同腺病毒载体。在早期区域2或4有缺失的载体在28天时表达β-半乳糖苷酶。目前为避免这种反应而设计的腺病毒载体在转基因持续时间方面的改善微乎其微。需要进一步对载体进行修饰或采用替代策略，以实现胰腺腺病毒转基因的稳定表达。
