Miyanohara A, Johnson P A, Elam R L, Dai Y, Witztum J L, Verma I M, Friedmann T
Department of Pediatrics, University of California, San Diego, La Jolla 92093-0634.
New Biol. 1992 Mar;4(3):238-46.
We have used gene transfer vectors derived from a replication-defective mutant of herpes simplex virus type 1 (HSV-1) expressing the hepatitis B virus surface antigen (HBsAg), Escherichia coli beta-galactosidase (beta-gal), or canine factor IX (cFIX) from the immediate early promoter of human cytomegalovirus (hCMV) to infect mouse liver by direct injection or through the portal vein. By either route, high levels of transgene expression were demonstrated by the detection of immunoreactive HBsAg or cFIX in the circulation and by histochemical detection of beta-gal activity in situ. The results were striking in that the serum level of cFIX reached 10% of the normal murine levels. Although the level of transgene expression from the hCMV promoter was transient, a significant number of persistent vectors could be rescued from the livers of recipient mice up to 2 months after inoculation. Replacement of the hCMV promoter with the HSV-1 latency-associated transcript (LAT) promoter resulted in reduced but prolonged expression of both HBsAg and cFIX. The very high level of factor IX expression suggests that clinically useful gene transfer may eventually be feasible through direct vector delivery to the liver.
我们使用了源自1型单纯疱疹病毒（HSV-1）复制缺陷型突变体的基因转移载体，该载体通过人巨细胞病毒（hCMV）的立即早期启动子表达乙型肝炎病毒表面抗原（HBsAg）、大肠杆菌β-半乳糖苷酶（β-gal）或犬因子IX（cFIX），通过直接注射或经门静脉感染小鼠肝脏。通过这两种途径，通过检测循环中的免疫反应性HBsAg或cFIX以及通过原位组织化学检测β-gal活性，证明了高水平的转基因表达。结果令人惊讶，因为cFIX的血清水平达到了正常小鼠水平的10%。尽管来自hCMV启动子的转基因表达水平是短暂的，但在接种后长达2个月的时间里，可以从受体小鼠的肝脏中拯救出大量的持久性载体。用HSV-1潜伏相关转录本（LAT）启动子取代hCMV启动子导致HBsAg和cFIX的表达降低但延长。因子IX的极高表达水平表明，通过直接将载体递送至肝脏，临床上有用的基因转移最终可能是可行的。
