远程载体递送后脊髓中腺病毒基因表达的特征分析。

Characterization of adenoviral gene expression in spinal cord after remote vector delivery.

作者信息

Boulis N M, Turner D E, Dice J A, Bhatia V, Feldman E L

机构信息

Section of Neurosurgery, University of Michigan, Ann Arbor, USA.

出版信息

Neurosurgery. 1999 Jul;45(1):131-7; discussion 137-8. doi: 10.1097/00006123-199907000-00029.

DOI:10.1097/00006123-199907000-00029

PMID:10414575

Abstract

OBJECTIVES

Recent work has established that the remote injection of attenuated adenoviral vectors may result in central nervous system (CNS) gene expression. These studies suggest that virus passes through peripheral nerves into the CNS. The present experiment attempts to characterize this phenomenon systematically.

METHODS

Spinal cord cells staining for the reporter gene beta-galactosidase were histologically quantified after microinjection of the viral vector Ad5RSVntLacZ into rat footpad, muscle, or sciatic nerve. The effects of injection location, titer, and time, as well as nerve crush and dexamethasone, were examined.

RESULTS

Sciatic nerve viral vector injection results in significantly higher CNS uptake than intramuscular and subcutaneous injections (P < 0.05). Nerve crush injury caused a time-dependent reduction in spinal cord gene uptake after sciatic nerve adenoviral injection (P < 0.05). Neuronal staining reaches its peak at 6 days after injection (P < 0.002). Peripheral nerve delivery to the CNS increases with augmented titers (P < 0.03). Finally, gene expression is augmented by administration of dexamethasone (P < 0.0001).

CONCLUSION

Remote adenoviral vector injection represents a potential method for spinal cord gene therapy that avoids any manipulation of CNS tissue.

摘要

目的

最近的研究证实，经外周注射减毒腺病毒载体可使中枢神经系统（CNS）实现基因表达。这些研究表明，病毒可通过外周神经进入中枢神经系统。本实验旨在系统地描述这一现象。

方法

将病毒载体Ad5RSVntLacZ显微注射到大鼠足垫、肌肉或坐骨神经后，对脊髓中表达报告基因β-半乳糖苷酶的细胞进行组织学定量分析。研究了注射部位、病毒滴度、时间以及神经挤压和地塞米松对实验结果的影响。

结果

与肌肉注射和皮下注射相比，坐骨神经注射病毒载体后中枢神经系统的摄取量显著更高（P < 0.05）。坐骨神经注射腺病毒后，神经挤压损伤导致脊髓基因摄取量随时间减少（P < 0.05）。神经元染色在注射后6天达到峰值（P < 0.002）。随着病毒滴度的增加，外周神经向中枢神经系统的基因传递量也增加（P < 0.03）。最后，地塞米松的给药可增强基因表达（P < 0.0001）。