A cystic fibrosis tracheal gland cell line, CF-KM4. Correction by adenovirus-mediated CFTR gene transfer.
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 DeltaF508 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 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.
Kammouni
W
W
Groupe de Recherche sur les Glandes Exocrines, Faculté de Médecine, Marseille, Cedex 05, France.
Moreau
B
B
Becq
F
F
Saleh
A
A
Pavirani
A
A
Figarella
C
C
Merten
M D
MD
D000818:Animals; D016466:CHO Cells; D002455:Cell Division; D002460:Cell Line; D002999:Clone Cells; D006224:Cricetinae; D003550:Cystic Fibrosis / Q000235:genetics / Q000473:pathology* / Q000503:physiopathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000096:biosynthesis* / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006720:Homozygote; D006801:Humans; D007633:Keratins / Q000032:analysis; D008564:Membrane Potentials; D011994:Recombinant Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D013539:Simian virus 40; D014132:Trachea / Q000473:pathology* / Q000503:physiopathology; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
C505032:CFTR protein, human; D011994:Recombinant Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D007633:Keratins; D001616:beta-Galactosidase