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 p ulmonary target for the gene therapy of cystic fibrosis (CF), We developed a CF tracheal gland serous cell line, CF-KM4, obtained by the transformatio n 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 retai ned epithelial and secretory features of the native CF-HTGS cells in primar y culture, namely, presence of cytokeratin, constitutive secretion of secre tory tory leukocyte proteinase inhibitor, absence of responsiveness to carb achol and isoproterenol, and defective cyclic adenosine monophosphate-depen dent chloride channel activity. Adenovirus-mediated CF transmembrane conduc tance regulator (CFTR) gene transfer into CF-KM4 cells corrected the defect ive chloride channel activity as well as the responsiveness to adrenergic a nd cholinergic agonists, In contrast, control transfection using adenovirus -mediated beta-galactosidase gene transfer was totally ineffective. In conc lusion, these results present a stable CF tracheal gland cell line that has retained its epithelial and CF-specific defective secretory characteristic s which are corrected after CFTR gene transfer. This cell Line therefore ap pears to be a useful tool for large-scale molecular and cellular pharmacolo gic investigations designed to test potential therapies of the disease CF.