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重复眼内腺病毒载体注射介导的病毒转基因表达:体内活体成像研究。

Viral transgene expression delivered by repeat intraocular adenoviral vector injection: in vivo live imaging study.

机构信息

Department of Ophthalmology, Johns Hopkins University, School of Medicine, Baltimore, MD, USA.

出版信息

Mol Imaging. 2012 Sep-Oct;11(5):361-71.

Abstract

We delivered adenovirus vector (Ad) via intravitreous injection and monitored transgene (luciferase) expression in living mice (BALB/c) at multiple time points. In vivo live imaging technology was able to assess dynamically intraocular luciferase expression in a single animal population throughout the entire experiment period. Using this information, we were able to determine the optimal time point for readministration of Ad into the eyes and to dynamically study the time course of expression of a second Ad administration. Optical imaging demonstrated the limited period of transgene expression in eyes. Significant transgene signal was also detected in livers. The repeat intraocular delivery of the adenovirus resulted in significant blunting of transgene expression in both eyes and livers compared to the initial delivery. Periocular corticosteroid (triamcinolone acetonide) injection combined with initial Ad delivery was effective to rescue luciferase expression on repeat Ad vector delivery. However, this effect was not observed when corticosteroid was combined with repeat Ad delivery. Although corticosteroid enhanced ocular transgene expression, it also increased transgene expression in liver, which has potential safety implications. This dynamic transgene expression in eyes was successfully traced and monitored via a live imaging technique.

摘要

我们通过玻璃体内注射将腺病毒载体(Ad)递送至活体小鼠(BALB/c)中,并在多个时间点监测活体小鼠中转基因(荧光素酶)的表达情况。活体实时成像技术能够在整个实验期间动态评估单个动物群体眼内荧光素酶的表达情况。利用这些信息,我们能够确定再次将 Ad 递送至眼内的最佳时间点,并动态研究第二次 Ad 给药的表达时间过程。光学成像显示了转基因在眼睛中的表达具有有限的时间周期。在肝脏中也检测到明显的转基因信号。与初次给药相比,重复眼内给予腺病毒导致眼睛和肝脏中转基因表达的显著减弱。与初次 Ad 给药联合使用球周皮质类固醇(曲安奈德)注射可有效挽救重复 Ad 载体给药时的荧光素酶表达。然而,当皮质类固醇与重复 Ad 给药联合使用时,未观察到这种效果。尽管皮质类固醇增强了眼部转基因的表达,但它也增加了肝脏中转基因的表达,这可能存在潜在的安全性问题。这种眼部的动态转基因表达可以通过活体成像技术成功追踪和监测。

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