Budenz D L, Bennett J, Alonso L, Maguire A
Scheie Eye Institute, F. M. Kirby Center for Molecular Ophthalmology, University of Pennsylvania School of Medicine, Philadelphia, USA.
Invest Ophthalmol Vis Sci. 1995 Oct;36(11):2211-5.
To determine whether a reporter gene can be introduced into adult mammalian corneal endothelial and trabecular meshwork cells in vivo using a recombinant replication-deficient adenovirus.
Purified replication-deficient adenovirus containing the cytomegalovirus-promoted Escherichia coli reporter gene, lacZ, was injected into the vitreous cavities or anterior chambers of 30 adult CD-1 mice using the contralateral eyes as controls. LacZ expression was assessed histochemically in enucleated eyes from 2 to 21 days after injection using the beta-Galactosidase (beta-Gal) assay.
LacZ expression was demonstrated in corneal endothelial and trabecular meshwork cells for as long as 14 days after injection. beta-Gal activity was also observed in lens and iris epithelial cells. There was no toxicity of the adenoviral vector demonstrated histologically, and no nonocular tissues expressed lacZ as measured by beta-Gal assay.
A functional gene can be transferred in vivo into adult mammalian corneal endothelial and trabecular meshwork cells using a replication-defective adenoviral vector. Gene expression is relatively short-lived compared to that demonstrated previously in other ocular tissues (photoreceptors and retinal pigment epithelium). Adenoviral vectors may be a viable means for short-term delivery of therapeutic genes in vivo to cells in the anterior segment of the eye.
确定是否可以使用重组复制缺陷型腺病毒将报告基因体内导入成年哺乳动物角膜内皮细胞和小梁网细胞。
将含有巨细胞病毒启动的大肠杆菌报告基因lacZ的纯化复制缺陷型腺病毒注入30只成年CD-1小鼠的玻璃体腔或前房,对侧眼作为对照。在注射后2至21天,使用β-半乳糖苷酶(β-Gal)测定法对摘除的眼球进行组织化学评估lacZ表达。
注射后长达14天,在角膜内皮细胞和小梁网细胞中均显示有lacZ表达。在晶状体和虹膜上皮细胞中也观察到β-Gal活性。组织学检查未显示腺病毒载体有任何毒性,通过β-Gal测定法检测也没有非眼组织表达lacZ。
使用复制缺陷型腺病毒载体可将功能性基因体内导入成年哺乳动物角膜内皮细胞和小梁网细胞。与先前在其他眼组织(光感受器和视网膜色素上皮)中显示的情况相比,基因表达持续时间相对较短。腺病毒载体可能是将治疗性基因体内短期递送至眼前节细胞的一种可行方法。