Sandig V, Hofmann C, Steinert S, Jennings G, Schlag P, Strauss M
HepaVec GmbH, Berlin-Buch, Germany.
Hum Gene Ther. 1996 Oct 20;7(16):1937-45. doi: 10.1089/hum.1996.7.16-1937.
Gene therapy of liver diseases requires the development of efficient vectors for gene transfer in vivo. Retroviral and adenoviral vectors have been shown to deliver genes efficiently into hepatocytes in vitro and in vivo. However, these vectors do not allow for exclusive infection of the liver which would be highly advantageous for in vivo gene therapy strategies. We have recently demonstrated that genetically modified baculoviruses (Autographa californica nuclear polyhedrosis virus) efficiently deliver genes into cultured cells and have a strong preference for hepatocytes of different origin. Baculoviral gene transduction efficiency into human hepatocytes was determined to approach 100% and expression levels are high, provided that gene expression is controlled by mammalian promoters. In this report, we present further properties of baculoviruses regarding their use for hepatocyte gene transfer. Baculovirus-mediated gene expression declines rapidly in the hepatocellular carcinoma cell line Huh7 and more slowly in primary cultures of mouse hepatocytes. Direct application of baculoviruses for gene delivery to the liver in vivo is hampered by serum components, presumably by complement. However, we demonstrate here that baculoviral gene transfer is feasible in ex vivo perfused human liver tissue. This result suggests the development of a strategy using baculoviral vectors for liver-directed gene therapy.
肝脏疾病的基因治疗需要开发高效的体内基因转移载体。逆转录病毒和腺病毒载体已被证明能在体外和体内将基因有效地传递到肝细胞中。然而，这些载体不能特异性地感染肝脏，而这对于体内基因治疗策略来说是非常有利的。我们最近证明，基因改造的杆状病毒（苜蓿银纹夜蛾核型多角体病毒）能有效地将基因传递到培养细胞中，并且对不同来源的肝细胞有很强的偏好性。只要基因表达由哺乳动物启动子控制，杆状病毒将基因转导到人类肝细胞中的效率可达到近100%，且表达水平很高。在本报告中，我们展示了杆状病毒在用于肝细胞基因转移方面的更多特性。杆状病毒介导的基因表达在肝癌细胞系Huh7中迅速下降，而在小鼠肝细胞原代培养物中下降得较慢。血清成分（可能是补体）阻碍了杆状病毒直接用于体内肝脏的基因传递。然而，我们在此证明杆状病毒基因转移在体外灌注的人类肝脏组织中是可行的。这一结果提示了一种使用杆状病毒载体进行肝脏定向基因治疗策略的开发。
