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 .