Activation of NF-kB mediates ICAM-1 induction in respiratory cells exposed to an adenovirus-derived vector.
Gene transfer to the respiratory tract by replication-deficient adenoviruses is limited by the induction of inflammatory and immune responses. We previously demonstrated that a E1-E3-deleted recombinant adenovirus carrying the expression cassette for the cystic fibrosis gene (Ad.CFTR) upregulates the expression of the pro-inflammatory intercellular adhesion molecule-1 (ICAM-1) both in vitro and in vivo. In the present work we suggest a role for the nuclear factor-kB (NF-kB) in Ad.CFTR-dependent up-regulation of ICAM-1 in respiratory epithelial A549 cells. Specifically, Ad.CFTR induced translocation of NF-kB into the nucleus and binding to the proximal -228/-218 NF-kB consensus sequence on the ICAM-1 promoter. Ad.CFTR also stimulated a 13-fold increase in NF-kB-dependent expression of the CAT reporter gene under the control of a region of the ICAM-1 promoter, including the proximal NF-kB consensus sequence. The Ad.CFTR-dependent increase of ICAM-1 mRNA was abolished by inhibitors of NF-kB, such as N-acetyl-L-cysteine, pyrrolidine dithiocarbamate, parthenolide and the synthetic peptide SN50. All these inhibitors abolished both Ad.CFTR-induced NF-kB DNA binding and transactivating activities. These results indicate a critical role of NF-kB in the pro-inflammatory response elicited by replication-deficient adenoviral vectors in respiratory cells.
Melotti
P
P
Laboratory of Molecular Pathology, Cystic Fibrosis Center, Verona, Italy.
Nicolis
E
E
Tamanini
A
A
Rolfini
R
R
Pavirani
A
A
Cabrini
G
G
D000111:Acetylcysteine / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics; D000975:Antioxidants / Q000494:pharmacology; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003550:Cystic Fibrosis / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000494:pharmacology; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D012717:Sesquiterpenes / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014178:Translocation, Genetic / Q000187:drug effects
D000975:Antioxidants; C505032:CFTR protein, human; D016328:NF-kappa B; D010455:Peptides; D011759:Pyrrolidines; D012333:RNA, Messenger; C118258:SN50 peptide; D012717:Sesquiterpenes; D013859:Thiocarbamates; D018799:Intercellular Adhesion Molecule-1; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C020972:pyrrolidine dithiocarbamic acid; C002669:parthenolide; D015500:Chloramphenicol O-Acetyltransferase; D000111:Acetylcysteine