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Dysregulated interleukin-8 secretion and NF-κB activity in human cystic fibrosis nasal epithelial cells  Salvatore Carrabino, Daniela Carpani, Alessandra.

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Presentation on theme: "Dysregulated interleukin-8 secretion and NF-κB activity in human cystic fibrosis nasal epithelial cells  Salvatore Carrabino, Daniela Carpani, Alessandra."— Presentation transcript:

1 Dysregulated interleukin-8 secretion and NF-κB activity in human cystic fibrosis nasal epithelial cells  Salvatore Carrabino, Daniela Carpani, Alessandra Livraghi, Maurizio Di Cicco, Diana Costantini, Elena Copreni, Carla Colombo, Massimo Conese  Journal of Cystic Fibrosis  Volume 5, Issue 2, Pages (May 2006) DOI: /j.jcf Copyright © 2006 European Cystic Fibrosis Society Terms and Conditions

2 Fig. 1 Basal and induced NF-κB activity in primary respiratory nasal and A549 cells. (A) Cells were either untreated or treated with IFN-γ (100 U/ml) for 60 min or TNF-α (10 ng/ml) for 30 min prior to the preparation of nuclear extracts. 2.5 μg of each nuclear extract were analyzed for κB-DNA binding activity by EMSA. In lanes 2, 4, 6, 8, 10, 12, and 15, a 50-fold excess of unlabeled NF-κB oligonucleotide was added to the reaction mixture. NF-κB complexes migrated slightly faster in A549 than in nasal cells. (B) Nasal cells (2 weeks-old) were either untreated or incubated with a clinical isolate of P. aeruginosa (PA; 5×104 cfu/ml) for 1 h. The arrows and the asterisks denote the specific band and the free probe, respectively. Journal of Cystic Fibrosis 2006 5, DOI: ( /j.jcf ) Copyright © 2006 European Cystic Fibrosis Society Terms and Conditions

3 Fig. 2 Time course of IL-8 secretion by non-CF and CF nasal outgrowths. Nasal polyps (non-CF=4; CF=5) were plated as 1–2 mm2 explants onto 60-mm plate dishes and conditioned medium was collected after 2, 5, 7, 10, and 15 days. ⁎P= CF vs. non-CF at each time point. Journal of Cystic Fibrosis 2006 5, DOI: ( /j.jcf ) Copyright © 2006 European Cystic Fibrosis Society Terms and Conditions

4 Fig. 3 Evaluation of NF-κB p65 nuclear activity by a colorimetric assay in CF and non-CF nasal epithelial cells under basal conditions and following exposure to P. aeruginosa PAO1 (PA; 1×108 cfu/ml) for 5 h (see Methods) or costimulation with IL-1β (30 ng/ml)/H2O2 (100 μM) for 5 h. ⁎P= PA vs. basal and IL-1β/H2O2 vs. basal; §P= basal CF vs. basal non-CF; #P= PA vs. basal and IL-1β/H2O2 vs. basal. Journal of Cystic Fibrosis 2006 5, DOI: ( /j.jcf ) Copyright © 2006 European Cystic Fibrosis Society Terms and Conditions

5 Fig. 4 IL-8 levels in supernatants of CF and non-CF nasal cells under basal condition and following exposure to P. aeruginosa PAO1 (PA; 1×108 cfu/ml) for 5 h (see Methods) or co-stimulation with IL-1β (30 ng/ml)/H2O2 (100 μM). ⁎P= PA vs. basal and IL-1β/H2O2 vs. basal; §P= basal CF vs. basal non-CF; #P=0.44 PA vs. basal; ⁎⁎P=0.61 IL-1β/H2O2 vs. basal. Journal of Cystic Fibrosis 2006 5, DOI: ( /j.jcf ) Copyright © 2006 European Cystic Fibrosis Society Terms and Conditions


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