Inhibition by TNF-alpha and IL-4 of cationic lipid mediated gene transfer in cystic fibrosis tracheal gland cells.
BACKGROUND
In vivo, tracheal gland serous cells highly express the cystic fibrosis transmembrane conductance regulator (cftr) gene. This gene is mutated in the lethal monogenic disease cystic fibrosis (CF). Clinical trials in which the human CFTR cDNA was delivered to the respiratory epithelia of CF patients have resulted in weak and transient gene expression.


RESULTS
As CF is characterized by mucus inspissation, airway infection, and severe inflammation, we tested the hypothesis that inflammation and especially two cytokines involved in the Th1/Th2 inflammatory response, interleukin 4 (IL-4) and TNFalpha, could inhibit gene transfer efficiency using a model of human CF tracheal gland cells (CF-KM4) and Lipofectamine reagent as a transfection reagent. The specific secretory defects of CF-KM4 cells were corrected by Lipofectamine-mediated human CFTR gene transfer. However, this was altered when cells were pre-treated with IL-4 and TNFalpha. Inhibition of luciferase reporter gene expression by IL-4 and TNFalpha pre-treated CF-KM4 cells was measured by activity and real-time RT-PCR. Both cytokines induced similar and synergistic inhibition of transgene expression and activity. This cytokine-mediated inhibition could be prevented by anti-inflammatory agents such as glucocorticoids but not by non-steroidal (NSAI) agents.


CONCLUSIONS
This data suggests that an inflammatory context generated by IL-4 and TNFalpha can inhibit human CFTR gene transfer in CF tracheal gland cells and that glucocorticoids may have a protecting action.
Laboratoire de Pathologie Cellulaire et Moléculaire en Nutrition, INSERM U724, Faculté de Médecine, BP 184, 54505 Vandoeuvre-lès-Nancy, France.
Sonia
S
Bastonero
Myriem
M
Gargouri
Sandrine
S
Ortiou
Jean-Louis
JL
Guéant
Marc D
MD
Merten
D000893:Anti-Inflammatory Agents / Q000378:metabolism; D002460:Cell Line; D003550:Cystic Fibrosis / Q000235:genetics / Q000276:immunology / Q000503:physiopathology / Q000628:therapy; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism; D003907:Dexamethasone / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism*; D008055:Lipids / Q000737:chemistry*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D014132:Trachea / Q000166:cytology* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D000893:Anti-Inflammatory Agents; D008055:Lipids; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D015847:Interleukin-4; D003907:Dexamethasone; D008156:Luciferases