Izembart A, Aguado E, Gauthier O, Aubert D, Moullier P, Ferry N
Laboratoire de Thérapie Génique, CHU Hôtel-Dieu, Nantes, France.
Hum Gene Ther. 1999 Dec 10;10(18):2917-25. doi: 10.1089/10430349950016339.
Gene transfer in regenerating dog liver using high-titer recombinant retroviral vectors carrying the E. coli beta-galactosidase gene was studied. Supernatants containing amphotropic or gibbon ape pseudotyped recombinant retroviruses were infused into a peripheral vein in beagle dogs after partial hepatectomy. The kinetics of liver regeneration were determined in the animals and daily infusions were carried out for 4 or 5 days during the regeneration period. Up to 2.8% of hepatocytes were beta-galactosidase positive at the end of the procedure. However, the number of positive cells declined rapidly and few positive hepatocytes were detected after 3 weeks. PCR demonstrated the disappearance of the provirus. Histologically, inflammatory lesions were observed in the transduced livers. Finally, we demonstrated the presence of a cytotoxic T lymphocyte immune response directed against beta-galactosidase-expressing cells, which could explain the disappearance of the transgene. This work suggests that the efficiency of in vivo gene delivery using high-titer retroviral vectors directly infused into the circulation may be hampered by a cytotoxic immune response against the infected cells.
利用携带大肠杆菌β-半乳糖苷酶基因的高滴度重组逆转录病毒载体，对再生犬肝脏中的基因转移进行了研究。部分肝切除术后，将含有嗜异性或长臂猿猿假型重组逆转录病毒的上清液注入比格犬的外周静脉。测定动物肝脏再生的动力学，并在再生期间每天进行4或5天的输注。在操作结束时，高达2.8%的肝细胞β-半乳糖苷酶呈阳性。然而，阳性细胞数量迅速下降，3周后几乎检测不到阳性肝细胞。PCR证明前病毒消失。组织学上，在转导的肝脏中观察到炎症病变。最后，我们证明了存在针对表达β-半乳糖苷酶细胞的细胞毒性T淋巴细胞免疫反应，这可以解释转基因的消失。这项工作表明，直接注入循环系统的高滴度逆转录病毒载体在体内基因传递的效率可能受到针对受感染细胞的细胞毒性免疫反应的阻碍。
