In vitro and in vivo transfer and expression of human surfactant SP-A- and SP-B-associated protein cDNAs mediated by replication-deficient, recombinant adenoviral vectors.
In vitro and in vivo transfer and expression of human surfactant SP-A- and SP-B-associated protein cDNAs mediated by replication-deficient, recombinant adenoviral vectors.
复制标题
由复制缺陷型重组腺病毒载体介导的人表面活性剂 SP-A 和 SP-B 相关蛋白 cDNA 的体外和体内转移和表达。
作者：
R. Korst;B. Bewig;R. Crystal
先天性肺泡蛋白沉积症（CPAP）是一种对目前的药物治疗无反应的足月婴儿的致命疾病。现在认识到，至少某些形式的这种疾病与SP-B（表面活性剂相关蛋白之一）的缺乏以及表面活性剂相关蛋白SP-A和SP-C的可能畸变有关。考虑到这些发展，合乎逻辑的假设是CPAP可能适合于基因治疗，其中人SP-B cDNA，以及可能的其他表面活性剂相关蛋白的cDNA，被转移到下呼吸道的上皮。我们构建了复制缺陷型重组腺病毒载体，其中组成型病毒启动子驱动表面活性剂相关蛋白SP-B（AdCMV.SP-B）和SP-A（AdCMV.SP-A）的DNA表达。在体外用这些载体感染人肺A549上皮细胞系后，检测到这些cDNA的适当大小的mRNA，而用对照病毒感染的细胞或未感染的细胞不产生mRNA。Western印迹证明了这些蛋白的表达，包括疏水蛋白SP-B的适当加工。在用这些载体体内肺内感染大鼠后，肺的北方分析揭示了这些cDNA的适当大小的mRNA，而用对照病毒感染的大鼠或未感染的大鼠显示不与人表面活性剂相关蛋白探针杂交。在AdCM-V.SP-A感染的大鼠中，Western印迹证实了与对照组相比，支气管肺泡灌洗液和肺匀浆中人SP-A蛋白的过量产生。因此，利用腺病毒载体在体外和体内转移和表达人表面活性物质相关蛋白cDNA是可行的，这为CPAP以及其他表面活性物质缺乏状态如新生儿呼吸窘迫综合征和可能的成人呼吸窘迫综合征提供了一种可能的治疗模式。
Congenital pulmonary alveolar proteinosis (CPAP) is a fatal disease of full-term infants that is unresponsive to current medical therapy. It is now recognized that at least some forms of this disorder are associated with a deficiency of SP-B, one of the surfactant-associated proteins, as well as probable aberrations in the surfactant-associated proteins SP-A and SP-C. Given these developments, it is logical to hypothesize that CPAP may be amenable to gene therapy, in which the human SP-B cDNA, and possibly the cDNAs of the other surfactant associated proteins, are transferred to the epithelium of the lower respiratory tract. We constructed replication-deficient, recombinant adenovirus vectors in which a constitutive viral promoter drives the expression of the DNAs for the surfactant-associated proteins, SP-B (AdCMV.SP-B) and SP-A (AdCMV.SP-A). Following infection of the human lung A549 epithelial cell line with these vectors in vitro, the appropriately sized mRNAs for these cDNAs were detected, whereas cells infected with a control virus or uninfected cells produced none. Western blots demonstrated expression of these proteins, including appropriate processing of the hydrophobic protein, SP-B. Following in vivo intratracheal infection of rats with these vectors, Northern analysis of the lungs revealed appropriately sized mRNAs for these cDNAs whereas rats infected with control virus or uninfected rats show no hybridization with the human surfactant-associated protein probes. In the AdCM-V.SP-A-infected rats, Western blots confirmed the overproduction of the human SP-A protein in both the bronchoalveolar lavage and lung homogenates compared to controls. Thus, it is feasible to utilize adenovirus vectors to transfer and express the human surfactant associated protein cDNAs in vitro and in vivo, presenting a possible mode of therapy for CPAP, as well as other surfactant deficiency states such as the neonatal respiratory distress syndrome and possibly the adult respiratory distress syndrome.
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DOI：
10.1164/ajrccm/147.3.653
发表时间：
1993-03-01
期刊：
AMERICAN REVIEW OF RESPIRATORY DISEASE
影响因子：
--
作者：
BAUGHMAN, RP;STERNBERG, RI;WHITSETT, J
通讯作者：
WHITSETT, J
影响因子：
15.9
作者：
HARTSHORN, KL;CROUCH, EC;SASTRY, K
通讯作者：
SASTRY, K
II 型肺泡细胞表达肺表面活性蛋白 A 的高亲和力受体。
DOI：
10.1073/pnas.85.15.5566
发表时间：
1988
影响因子：
11.1
作者：
Kuroki,Y;Mason,RJ;Voelker,DR
通讯作者：
Voelker,DR
表面活性剂脱辅基蛋白 Mr = 26,000-36,000 增强 II 型细胞对脂质体的摄取。
DOI：
--
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Wright,JR;Wager,RE;Hawgood,S;Dobbs,L;Clements,JA
通讯作者：
Clements,JA
使用人表面活性剂低分子量脱辅基蛋白重建表面活性剂生物活性。
DOI：
10.1172/jci113391
发表时间：
1988
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Revak,SD;Merritt,TA;Degryse,E;Stefani,L;Courtney,M;Hallman,M;Cochrane,CG
通讯作者：
Cochrane,CG