Rosenfeld M A, Yoshimura K, Trapnell B C, Yoneyama K, Rosenthal E R, Dalemans W, Fukayama M, Bargon J, Stier L E, Stratford-Perricaudet L
Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
Cell. 1992 Jan 10;68(1):143-55. doi: 10.1016/0092-8674(92)90213-v.
Direct transfer of the normal cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene to airway epithelium was evaluated using a replication-deficient recombinant adenovirus (Ad) vector containing normal human CFTR cDNA (Ad-CFTR). In vitro Ad-CFTR-infected CFPAC-1 CF epithelial cells expressed human CFTR mRNA and protein and demonstrated correction of defective cAMP-mediated Cl- permeability. Two days after in vivo intratracheal introduction of Ad-CFTR in cotton rats, in situ analysis demonstrated human CFTR gene expression in lung epithelium. PCR amplification of reverse transcribed lung RNA demonstrated human CFTR transcripts derived from Ad-CFTR, and Northern analysis of lung RNA revealed human CFTR transcripts for up to 6 weeks. Human CFTR protein was detected in epithelial cells using anti-human CFTR antibody 11-14 days after infection. While the safety and effectiveness remain to be demonstrated, these observations suggest the feasibility of in vivo CFTR gene transfer as therapy for the pulmonary manifestations of CF.
使用包含正常人囊性纤维化跨膜传导调节因子（CFTR）cDNA的复制缺陷型重组腺病毒（Ad）载体（Ad-CFTR），评估了正常CFTR基因向气道上皮的直接转移。体外Ad-CFTR感染的CFPAC-1囊性纤维化上皮细胞表达人CFTR mRNA和蛋白，并显示出缺陷性cAMP介导的Cl-通透性得到纠正。在棉鼠体内气管内导入Ad-CFTR两天后，原位分析显示肺上皮中有人类CFTR基因表达。对逆转录的肺RNA进行PCR扩增，证明了源自Ad-CFTR的人类CFTR转录本，对肺RNA的Northern分析显示人类CFTR转录本可持续6周。感染后11-14天，使用抗人CFTR抗体在上皮细胞中检测到了人CFTR蛋白。虽然安全性和有效性仍有待证明，但这些观察结果表明体内CFTR基因转移作为治疗CF肺部表现的可行性。
