Adenovirus-mediated gene transfer to glomerular cells in newborn mice
Adenovirus-mediated gene transfer to glomerular cells in newborn mice
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DOI：
10.1007/s00467-005-1882-0
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发表时间：
2005-10-01
影响因子：
3
通讯作者：
Ray, PE
中科院分区：
文献类型：
--
作者：
Jerebtsova, M;Liu, XH;Ray, PE
由于循环病毒被肝脏快速清除，重组腺病毒（rAd）载体向肾小球的全身递送一直是有问题的。我们以前已经表明，通过肝旁路延长rAd载体在循环中的保留改善了成年小鼠和大鼠中肾小球细胞的转导。进行本研究以确定新生小鼠在全身注射rAd载体后是否具有来自循环的rAd载体的延迟清除以及肾小球细胞的更有效转导。通过眶后静脉丛（2 × 109个颗粒/g体重）向新生（1天）和成年（6个月）C57 B16/J小鼠（每组n =20）注射携带lacZ基因的rAd载体（rAd.lacZ）。通过Western印迹、免疫组织化学、β-半乳糖苷酶染色、酶活性测定和RT-PCR研究，在新生和成年小鼠的不同时间点评估科萨基和腺病毒受体（CAR）和lacZ基因的表达。rAd.lacZ在新生小鼠体内的清除率明显延迟，这些小鼠体内的循环病毒浓度几乎是成年小鼠的10倍。新生儿肾脏显示CAR表达增加，主要位于肾小球细胞中。这些发现与lacZ基因向新生小鼠肾小球和肾小管的有效基因转移有关。本研究首次证明了使用全身静脉注射rAd载体在发育中的幼鼠肾小球中表达外源基因的可行性。
The systemic delivery of recombinant adenoviral (rAd) vectors to renal glomeruli has been problematic due to the rapid clearance of the circulating virus by the liver. We have previously shown that prolonged retention of rAd vectors in the circulation by liver bypass improves the transduction of renal glomerular cells in adult mice and rats. This study was done to determine whether newborn mice have a delayed clearance of rAd vectors from the circulation and a more efficient transduction of glomerular cells after a systemic injection of rAd vectors. Newborn (1 day old) and adult (6 months old) C57Bl6/J mice ( n =20 in each group) were injected with rAd vectors carrying the lacZ gene (rAd.lacZ) through the retro-orbital venous plexus (2x10(9) particles/g body weight). The renal expression of Coxsackie and Adenoviral Receptors (CAR) and lacZ gene were evaluated at different time points by Western blots, immunohistochemistry, beta-galactosidase staining, enzyme assay activity, and RT-PCR studies in newborn and adult mice. The clearance rate of rAd.lacZ was significantly delayed in newborn mice, and the concentration of circulating virus in these mice was almost ten times higher than that in adult mice. Newborn kidneys showed increased expression of CAR, predominately localized in glomerular cells. These findings were associated with an efficient gene transfer of the lacZ gene into glomeruli and tubules of newborn mice. This study demonstrates for the first time the feasibility of using systemic intravenous injections of rAd vectors to express foreign genes in developing glomeruli of young mice.