Exogenous surfactant enhances the delivery of recombinant adenoviral vectors to the lung
Exogenous surfactant enhances the delivery of recombinant adenoviral vectors to the lung
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DOI：
10.1089/hum.1997.8.2-171
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发表时间：
1997-01-20
影响因子：
4.2
通讯作者：
Welty, SE
中科院分区：
文献类型：
--
作者：
Katkin, JP;Husser, RC;Welty, SE
肺部疾病的体细胞基因治疗必须在体内完成，这需要基因转移载体在大面积的呼吸上皮上传播。表面活性剂是一种天然存在的蛋白质和脂质混合物，用于治疗早产儿呼吸窘迫综合征，在整个肺部迅速均匀地分散。我们使用外源性牛表面活性剂（Survanta beractant）作为肺传递表达β -半乳糖苷酶（β -gal）的重组腺病毒的载体。用腺病毒-滤除剂混合物处理的大鼠比用等量盐水处理的大鼠表现出更均匀的转基因表达的叶状分布。通过与邻硝基酚β - d -半乳糖苷（ONPG）反应，检测组织匀浆中β -gal的定量表达。β - gal活性的程度受用于传递病毒的载体的体积和类型的影响。在低体积（0.5 ml, 1.3 ml/kg）下，与盐水处理的动物和未处理的对照组相比，生物活性剂处理的动物表现出显著更高的肺β - gal活性（p < 0.002）。在高容量（1.2 ml, 4 ml/kg）下，平均β - gal活性在用脱硝剂或生理盐水处理组之间相似，但在生理盐水处理组内变化更大。无论使用何种载体，更高体积的传递介质都与β - gal表达水平的增加有关。Survanta对动物的耐受性良好，不影响转基因表达的持续时间。外源性分离剂为将重组腺病毒传递到肺部提供了一种有用的介质，当需要基因表达的弥散分布时。
Somatic gene therapy for pulmonary diseases must be accomplished in vivo, requiring the spread of a gene transfer vector across a vast expanse of respiratory epithelium. Surfactant, a naturally occurring protein and lipid mixture used to treat the respiratory distress syndrome of prematurity, disperses rapidly and evenly throughout the lung. We employed exogenous bovine surfactant (Survanta beractant) as a carrier vehicle for pulmonary delivery of a recombinant adenovirus expressing beta-galactosidase (beta-Gal). Rats treated with an adenovirus-beractant mixture demonstrated more uniform lobar distribution of transgene expression than rats treated with the same amount of virus in saline. Tissue homogenates were examined for quantitative beta-Gal expression by reaction with o-nitrophenol beta-D-galactopyranoside (ONPG). The degree of beta-Gal activity was affected by both the volume and type of carrier used to deliver the virus. At low volumes (0.5 ml, 1.3 ml/kg), beractant-treated animals demonstrated significantly greater pulmonary beta-Gal activity than saline-treated animals (p < 0.002) and untreated controls. At high volume (1.2 ml, 4 ml/kg), average beta-Gal activity was similar between groups treated with beractant or saline, but was more variable within the saline treated group. Higher volumes of delivery medium were associated with increased levels of beta-Gal expression regardless of the carrier used. Survanta was well tolerated by the animals and did not affect the duration of transgene expression. Exogenous beractant provides a useful medium for delivering recombinant adenoviruses to the lung when diffuse distribution of transgene expression is desired.