Cytoplasmic deposition of NFkappaB decoy oligonucleotides is insufficient to inhibit bleomycin-induced pulmonary inflammation.
Lung inflammation leads to severe tissue destruction and ultimately organ failure in a number of diseases, including cystic fibrosis (CF). The transcription factor nuclear factor kappa B (NFkappaB) regulates expression of many pro-inflammatory mediators. We have assessed the effect of topical administration of NFkappaB decoys in a bleomycin model of acute lung inflammation. Using fluorescein-labelled decoy oligonucleotides (ODN) (80 microg/mouse) we have shown that lipid-complexed and 'naked' ODN transfect conducting airway epithelium in a comparable manner (approximately 65% of cells). However, the ODN were detectable in the cytoplasm, but not in the nucleus of transfected cells. An increase of ODN dose to 500 microg/mouse did not increase nuclear transfection significantly. We determined the effect of cytoplasmic NFkappaB decoys on bleomycin-induced inflammation. We transfected mice with 'naked' decoy and scrambled ODN (500 microg) 1 h before intratracheal administration of bleomycin. We measured IL6 secretion in BALF and lung homogenates and total and differential cell counts in BALF 5 days after bleomycin administration. We did not detect a difference between NFkappaB decoy and scrambled ODN-treated animals in any of the parameters tested. We suggest that access of ODN to the nucleus of airway epithelial cells is a key problem, limiting the efficacy of such decoy strategies, as well as attempts at gene repair.
Department of Gene Therapy, Faculty of Medicine, Imperial College, London, UK.
U
U
Griesenbach
R L
RL
Cassady
R J
RJ
Cain
R M
RM
duBois
D M
DM
Geddes
E W F W
EW
Alton
D000818:Animals; D001761:Bleomycin; D001992:Bronchoalveolar Lavage Fluid / Q000276:immunology; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D015850:Interleukin-6 / Q000096:biosynthesis; D008168:Lung / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D018716:Molecular Mimicry; D016328:NF-kappa B / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics / Q000493:pharmacokinetics*; D011014:Pneumonia / Q000139:chemically induced / Q000276:immunology / Q000517:prevention & control*; D014018:Tissue Distribution; D014162:Transfection
D015850:Interleukin-6; D016328:NF-kappa B; D009841:Oligonucleotides; D001761:Bleomycin