Messina E, Muhlhauser J, Giuliano M, Pandolfi A, Morgese G, Procopio A
Experimental Center on Gene Therapy and Diagnosis, Gabriele D'Annunzio University, Chieti, Italy.
Pediatr Res. 1996 Jul;40(1):142-7. doi: 10.1203/00006450-199607000-00024.
Local delivery of Escherichia coli beta-galactosidase gene (beta-gal) to surfactant protein-A (SP-A)-producing cells by a replication-defective recombinant adenovirus (AdCMV.beta-gal) was tested in human 8-12-wk-old fetal lung explants cultured in Waymouth's medium. In contrast to uninfected explants, direct addition of 0.8-1.6 x 10(6) plaque-forming units of AdCMV.beta-gal resulted in beta-galactosidase (beta-Gal)-specific staining of the pulmonary epithelium. SP-A localization by indirect immunofluorescence showed positive specific staining of the beta-Gal+ lung epithelial cells, demonstrating that recombinant-defective adenoviruses efficiently transfer reporter genes to fetal lung SP-A+ cells. The reporter gene expression in SPA+ cells persisted for more than 1 mo. No apparent alteration of morphology, phenotype, and growth was observed. The in vitro human lung model described may be useful for testing DNA constructs for vector-mediated gene therapy, as an approach to the treatment of congenital defects and neonatal disorders, such as respiratory distress syndrome and bronchopulmonary dysplasia.
在Waymouth培养基中培养的人8至12周龄胎儿肺外植体中，测试了通过复制缺陷型重组腺病毒（AdCMV.β-gal）将大肠杆菌β-半乳糖苷酶基因（β-gal）局部递送至产生表面活性蛋白A（SP-A）的细胞。与未感染的外植体相比，直接添加0.8 - 1.6×10⁶个空斑形成单位的AdCMV.β-gal导致肺上皮细胞出现β-半乳糖苷酶（β-Gal）特异性染色。通过间接免疫荧光进行的SP-A定位显示β-Gal⁺肺上皮细胞有阳性特异性染色，表明重组缺陷型腺病毒能有效地将报告基因转移至胎儿肺SP-A⁺细胞。报告基因在SP-A⁺细胞中的表达持续超过1个月。未观察到形态、表型和生长有明显改变。所描述的体外人肺模型可能有助于测试用于载体介导基因治疗的DNA构建体，作为治疗先天性缺陷和新生儿疾病（如呼吸窘迫综合征和支气管肺发育不良）的一种方法。
