title: Sustained inhibition of IL-6 and IL-8 expression by decoy ODN to NF-κB delivered through respirable large porous particles in LPS-stimulated cystic fibrosis bronchial cells.
abstract: BACKGROUND
  Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Neutrophil-dominated inflammation and chronic bacterial infection are still considered the primary cause of bronchioectasis, respiratory failure and consequent death in CF patients. Activation of nuclear factor (NF)-κB is responsible for overproduction of cytokines, such as interleukin (IL)-6 and IL-8, in airways of CF patients. Thus, decoy oligodeoxynucleotides against NF-κB (dec-ODN) may limit lung inflammation in CF. In the present study, we studied the effects of dec-ODN delivered through biodegradable and respirable poly(D,L-lactide-co-glycolide) large porous particles (LPP) on IL-6 and IL-8 mRNA expression as well as NF-κB/DNA binding activity in cystic fibrosis cells stimulated with lipopolysaccharide (LPS) from Pseudomonas aeruginosa.
  
  
  METHODS
  dec-ODN LPP were prepared by a modified double emulsion technique and characterized in terms of size, morphology, tapped density and dec-ODN loading. Human epithelial bronchial IB3-1 (CFTR-mutated) as well as S9 (CFTR-corrected) were stimulated with LPS from P. aeruginosa for 24 and 72 h in the absence or presence of naked dec-ODN or dec-ODN LPP.
  
  
  RESULTS
  Stimulation of cells with LPS from P. aeruginosa caused an increase of IL-6 and IL-8 mRNA levels, which were significantly inhibited by dec-ODN LPP at 24 and 72 h, whereas naked dec-ODN inhibited those only at 24 h. Similar effects were exhibited by dec-ODN LPP or naked dec-ODN on NF-κB/DNA binding activity.
  
  
  CONCLUSIONS
  Our observations indicate that respirable biodegradable dec-ODN LPP may represent a promising strategy for inhibiting NF-κB transcriptional activity and related gene expression and, thus, reduce lung chronic inflammation in CF patients.
authors.affiliation: Department of Experimental Pharmacology, School of Biotechnological Sciences, University of Naples Federico II, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: De Stefano
authors.forename: Francesca
authors.initials: F
authors.lastname: Ungaro
authors.forename: Concetta
authors.initials: C
authors.lastname: Giovino
authors.forename: Antonio
authors.initials: A
authors.lastname: Polimeno
authors.forename: Fabiana
authors.initials: F
authors.lastname: Quaglia
authors.forename: Rosa
authors.initials: R
authors.lastname: Carnuccio
mesh_terms: D000704:Analysis of Variance; D001980:Bronchi / Q000166:cytology* / Q000276:immunology; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000276:immunology*; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000276:immunology; D006801:Humans; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism*; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism*; D008070:Lipopolysaccharides; D016328:NF-kappa B / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000008:administration & dosage / Q000494:pharmacology*
chemical_list: D017931:DNA Primers; D015850:Interleukin-6; D016209:Interleukin-8; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D014157:Transcription Factors