Gene transfer to the rat kidney in vivo and ex vivo using an adenovirus vector:: factors influencing transgene expression
Gene transfer to the rat kidney in vivo and ex vivo using an adenovirus vector:: factors influencing transgene expression
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DOI：
10.1093/ndt/gfh783
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发表时间：
2005-07-01
影响因子：
6.1
通讯作者：
Kobayashi, E
中科院分区：
文献类型：
--
作者：
Fujishiro, J;Takeda, SI;Kobayashi, E
背景腺病毒介导的基因转移到肾脏的特点还没有得到很好的研究。我们研究了接触时间和温度对腺病毒介导的转基因表达在大鼠肾脏中的影响，使用基于导管的体内基因转移和大鼠肾移植模型离体。通过肾动脉导管将含有荧光素酶（Ad-Luc）或β-半乳糖苷酶（Ad-LacZ）基因的腺病毒载体在体内引入肾中。评价了各种接触时间和温度。离体，肾移植物通过肾动脉注射Ad-Luc，冷却60分钟，然后移植。通过非侵入性生物成像系统或组织学定期评价荧光素酶表达。通过免疫电镜鉴定表达LacZ基因的细胞。在体内基因转移中，成功实现了转基因表达;然而，其效率与接触时间或温度无关。在离体基因转移中，移植肾中的转基因表达早期达到峰值并逐渐下降。在受体的肝脏中观察到强基因表达。LacZ在成纤维细胞、Bowman囊壁上皮细胞、系膜细胞、足细胞和肾小管细胞中均有表达。这项研究产生了有关体内和离体基因转移到肾脏的新信息，这将对肾脏基因治疗有用。
Background. The characteristics of adenovirus-mediated gene transfer into the kidney are not well examined. We studied the effects of contact time and temperature on adenovirus-mediated transgene expression in rat kidneys, using catheter-based in vivo gene transfer and a rat renal transplant model ex vivo.Methods. An adenovirus vector containing the luciferase (Ad-Luc) or beta-galactosidase (Ad-LacZ) gene was introduced in vivo into the kidney via a renal artery catheter. Various contact times and temperatures were evaluated. Ex vivo, the renal graft was injected with Ad-Luc through the renal artery, chilled for 60 min and then transplanted. Luciferase expression was evaluated periodically by a non-invasive bioimaging system or histology. Cells expressing the LacZ gene were identified by immunoelectron microscopy.Results. In in vivo gene transfer, successful transgene expression was achieved; however, its efficiency was independent of contact time or temperature. In ex vivo gene transfer, transgene expression in the renal graft peaked early and gradually decreased. Strong gene expression was observed in the recipients' livers. LacZ expression was detected in fibroblasts, parietal epithelial cells of Bowman's capsule, mesangial cells, podocytes and tubular cells.Conclusions. This study generated new information about in vivo and ex vivo gene transfer into the kidney, which would be useful for renal gene therapy.