Ohmori Kenichi, Takeda Shin-ichi, Miyoshi Shinichiro, Minami Masato, Nakane Shigeru, Ohta Mitsunori, Sawa Yoshiki, Matsuda Hikaru
Department of Surgery (E1), Osaka University Graduate School of Medicine, Yamada-oka 2-2, Suita City, Osaka 565-0871, Japan.
Eur J Cardiothorac Surg. 2005 May;27(5):768-73. doi: 10.1016/j.ejcts.2004.12.055.
A novel hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer system has been shown to have benefits of a high efficiency of transfection and low immunogenicity. The aims of this study were to determine the effect of re-transfection of the HVJ-liposome system via the airway, and to quantify the distribution of gene expression between transtracheal and transplantation approaches.
Beta-galactosidase (beta-gal) plasmid DNA was introduced into lung tissues using the HVJ-liposome method. Two groups of Sprague-Dawley (SD) rats received intratracheal instillation of 10 microg of the beta-gal gene, once on Day 0 in 1 group (Group Tb-1, n=4) and 3 times on Days 0-2 in another (Group Tb-3, n=4). In a third group of SD rats (Group Tx, n=5), an orthotopic left lung transplantation was performed after the donor lung was flushed with an HVJ-liposome complex solution and preserved for 1h. Gene expression and distribution in lung tissue was then quantified by counting the X-gal stained cells.
Both the transtracheal and transplantation approaches resulted in low levels of transfection in the vascular endothelial cells (0.2+/-0.1 and 4.0+/-1.8%), respectively, but a moderate degree of transfection to the airway (11.0+/-7.1 and 28.0+/-20.7%) and alveolar cells (3.0+/-1.8 and 6.0+/-3.6%). Three repetitive injections via the airway increased gene expression in airway epithelial cells of 41.0+/-12.0% compared with the single administration of 11.0+/-4.3%.
Our results suggest that the repeated transtracheal gene transfection using HVJ-liposome may have benefits for treatment of problems after lung transplantation. In addition, gene transfer using a flushing solution during harvest may provide an opportunity for gene manipulation in the setting of lung transplantation.
一种新型的日本血凝病毒（HVJ）-脂质体介导的基因转移系统已显示出具有转染效率高和免疫原性低的优点。本研究的目的是确定通过气道再次转染HVJ-脂质体系统的效果，并量化经气管和移植途径之间基因表达的分布。
使用HVJ-脂质体方法将β-半乳糖苷酶（β-gal）质粒DNA导入肺组织。两组Sprague-Dawley（SD）大鼠接受气管内滴注10μgβ-gal基因，一组（Tb-1组，n = 4）在第0天滴注一次，另一组（Tb-3组，n = 4）在第0 - 2天滴注3次。在第三组SD大鼠（Tx组，n = 5）中，在供体肺用HVJ-脂质体复合溶液冲洗并保存1小时后进行原位左肺移植。然后通过计数X-gal染色细胞来量化肺组织中的基因表达和分布。
经气管和移植途径分别导致血管内皮细胞的低水平转染（0.2±0.1和4.0±1.8%），但对气道（11.0±7.1和28.0±20.7%）和肺泡细胞（3.0±1.8和6.0±3.6%）有中等程度的转染。与单次给药的11.0±4.3%相比，通过气道进行三次重复注射使气道上皮细胞中的基因表达增加了41.0±12.0%。
我们的结果表明，使用HVJ-脂质体进行重复经气管基因转染可能对肺移植后问题的治疗有益。此外，在收获期间使用冲洗溶液进行基因转移可能为肺移植中的基因操作提供机会。
