Effect of exogenous nerve growth factor on neurotoxicity of and neuronal gene delivery by a herpes simplex amplicon vector in the rat brain.
Effect of exogenous nerve growth factor on neurotoxicity of and neuronal gene delivery by a herpes simplex amplicon vector in the rat brain.
复制标题
外源性神经生长因子对大鼠脑中单纯疱疹病毒扩增子载体的神经毒性和神经元基因传递的影响。
DOI：
10.1089/hum.1994.5.8-987
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发表时间：
1994
影响因子：
4.2
通讯作者：
Isacson,O
中科院分区：
文献类型：
--
作者：
Pakzaban,P;Geller,AI;Isacson,O
我们以前已经表明，局部破坏的神经组织的野生型单纯疱疹病毒1型（HSV-1）是减弱脑内输注神经生长因子（NGF）。为了研究NGF对HSV-1扩增子载体系统介导的神经溶解程度和体内神经元基因转移效率的影响，将扩增子制剂pHSVlac立体定位注射到大鼠纹状体中。该扩增子包含在HSV-1立即早期4/5启动子的转录控制下的大肠杆菌lacZ基因，并由携带立即早期3基因缺失的HSV-1辅助病毒包装。载体注射后连续脑内输注NGF-β（总剂量5 μg）或载体溶液7天。在7天输注期结束时处死动物，对脑进行组织学分析。在所有接种载体的动物中，注射部位周围有明显的炎症和坏死区。神经生长因子处理的动物的纹状体组织破坏体积（1.27 ± 0.19 mm 3;平均值± SEM）显著小于赋形剂处理的对照组（2.16 ± 0.37 mm 3;P< 0.05字节检验）。溶剂处理动物中HSV和β-半乳糖苷酶（β-Gal）的免疫组织化学染色显示，许多纹状体细胞携带HSV抗原（3，678 ± 636），但在注射后7天仅少数细胞表达报告基因（294 ± 60）。此时，NGF输注未显著影响HSV免疫反应细胞的数量（4，224 ± 618）或表达β-Gal的细胞数量（330 ± 72）。在两组的一些动物中，在整个大脑中观察到HSV免疫反应性和报告基因表达的弥散模式。我们的结论是外源性神经生长因子减少了这种HSV-1衍生载体在大鼠纹状体的局部细胞病理学效应，但不影响HSV免疫反应细胞的数量或转基因的短期表达。
We have previously shown that local destruction of neural tissue by wild-type herpes simplex virus type 1 (HSV-1) is attenuated by intracerebral infusion of nerve growth factor (NGF). To investigate the effect of NGF on the extent of neurolysis and efficacy of neuronal gene transfer mediated by an HSV-1 amplicon vector systemin vivo, rats were stereotaxically injected in the striatum with an amplicon preparation, pHSVlac. This amplicon contains theEscherichia coli lacZgene under the transcriptional control of the HSV-1 immediate early 4/5 promoter and is packaged by an HSV-1 helper virus carrying a deletion in the immediate early 3 gene. Vector injection was followed by continuous intracerebral infusion of NGF-β (total dose 5 μg) or vehicle solution over 7 days. Animals were sacrificed at the end of the 7-day infusion period for histological analysis of the brains.A distinct zone of inflammation and necrosis surrounded the injection site in all vector-inoculated animals. The volume of striatal tissue destruction was significantly smaller in NGF-treated animals (1.27 ± 0.19 mm3; mean ± SEM) than in the vehicle-treated controls (2.16 ± 0.37 mm3;P< 0.05 byt-test). Immunohistochemical staining for HSV and β-galactosidase (β-Gal) in vehicle-treated animals revealed that many striatal cells harbored HSV antigens (3,678 ± 636), but only a small number expressed the reporter gene at 7 days post-injection (294 ± 60). NGF infusion did not significantly affect the number of HSV-immunoreactive cells (4,224 ± 618), or the number of cells expressing β-Gal (330 ± 72) at this time. In some animals in both groups, a disseminated pattern of HSV immunoreactivity and reporter gene expression was seen throughout the brain. We conclude that exogenous NGF reduces the local cytopathologic effects of this HSV-1-derived vector in the rat striatum, but does not affect the number of HSV-immunoreactive cells or the short-term expression of the transgene.