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Am. J. Respir. Cell Mol. Biol., Volume 20, Number 4, April, 1999 684-691

A Cystic Fibrosis Tracheal Gland Cell Line, CF-KM4
Correction by Adenovirus-Mediated CFTR Gene Transfer

Wafa Kammouni, Bertrand Moreau,* Frederic Becq,* Ali Saleh, Andrea Pavirani, Catherine Figarella, and Marc D. Merten

Groupe de Recherche sur les Glandes Exocrines, Faculté de Médecine, Marseille; Laboratoire de Physiologie des Éléments Excitables, Université Claude Bernard, Villeurbanne; Laboratoire de Neurobiologie, Marseille; and Transgene, Strasbourg, France

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 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.

* Permanent address: Laboratorie de Physiologie des Regulations Cellulaires, Universite de Poitiers, UMR 6558, 40 Av du recteur Pineau, 86022 Poitiers, France.




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