A cystic fibrosis tracheal gland cell line, CF-KM4 - Correction by adenovirus-mediated CFTR gene transfer

被引:36
作者
Kammouni, W
Moreau, B
Becq, F
Saleh, R
Pavirani, A
Figarella, C
Merten, MD
机构
[1] Fac Med Marseille, Grp Rech Glandes Exocrines, F-13385 Marseille 05, France
[2] Univ Lyon 1, Lab Physiol Elements Excitables, F-69622 Villeurbanne, France
[3] Neurobiol Lab, Marseille, France
[4] Transgene SA, Strasbourg, France
[5] Univ Poitiers, Lab Physiol Regulat Cellulaires, UMR 6558, F-86022 Poitiers, France
关键词
D O I
10.1165/ajrcmb.20.4.3341
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human tracheal gland serous (HTGS) cells are now considered one principal pulmonary target for the gene therapy of cystic fibrosis (CF), We developed a CF tracheal gland serous cell line, CF-KM4, obtained by the transformation of primary cultures of CF tracheal gland serous cells homozygous for the Delta F508 mutation by using the wild-type SV40 virus. This cell line retained epithelial and secretory features of the native CF-HTGS cells in primary culture, namely, presence of cytokeratin, constitutive secretion of secretory tory leukocyte proteinase inhibitor, absence of responsiveness to carbachol and isoproterenol, and defective cyclic adenosine monophosphate-dependent chloride channel activity. Adenovirus-mediated CF transmembrane conductance regulator (CFTR) gene transfer into CF-KM4 cells corrected the defective chloride channel activity as well as the responsiveness to adrenergic and cholinergic agonists, In contrast, control transfection using adenovirus-mediated beta-galactosidase gene transfer was totally ineffective. In conclusion, these results present a stable CF tracheal gland cell line that has retained its epithelial and CF-specific defective secretory characteristics which are corrected after CFTR gene transfer. This cell Line therefore appears to be a useful tool for large-scale molecular and cellular pharmacologic investigations designed to test potential therapies of the disease CF.
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收藏
页码:684 / 691
页数:8
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