Submitted on May 17, 2007
Revised on February 1, 2008
Transcription Factor Oligodeoxynucleotides to NF-
B Inhibit Transcription of IL-8 in Bronchial Cells
Valentino Bezzerri1, Monica Borgatti2, Elena Nicolis1, Ilaria Lampronti2, Maria Cristina Dechecchi1, Irene Mancini2, Paolo Rizzotti1, Roberto Gambari3, and Giulio Cabrini1*
1 University-Hospital, Laboratory of Molecular Pathology, Laboratory of Clinical Chemistry and Haematology, Verona, Italy,
2 Department of Biochemistry and Molecular Biology, University of Ferrara, ER-GenTech, Ferrara, Italy,
3 Department of Biochemistry and Molecular Biology, University of Ferrara, ER-GenTech, Ferrara, Italy; University of Ferrara, Interdisciplinary Center for the Study of Inflammation, Ferrara, Italy
* To whom correspondence should be addressed. E-mail: giulio.cabrini{at}azosp.vr.it.
Chronic pulmonary inflammation in patients affected by cystic fibrosis (CF) is characterized by massive bronchial infiltrates of neutrophils, which is sustained by the interaction of pathogens (e.g. Pseudomonas aeruginosa) with surface bronchial cells. To explore new treatment options focused on the reduction of neutrophil chemotaxis, we applied the transcription factor (TF) decoy approach, based on the intracellular delivery of double stranded oligodeoxynucleotides (ODNs) causing inhibition of the binding of TF-related proteins to the different consensus sequences in the promoter of specific genes. In CF bronchial IB3-1 cells, P.aeruginosa induced transcription of the neutrophil chemokines IL-8 and GRO-
of the adhesion molecule ICAM-1, and of the cytokines IL-1
and IL-6. Since consensus sequences for the TF, NF-
B, is contained in the promoters of all these genes, IB3-1, CuFi-1, Beas-2B and CaLu-3 cells were transfected with double stranded TF "decoy" ODNs mimicking different NF-B consensus sequences. IL-8 NF-B decoy ODN partially inhibited the P.aeruginosa-dependent transcription of IL-8, GRO- and IL-6, whereas decoy ODNs to both HIV-1 LTR and Igk produced a strong, 80 to 85% inhibition of transcription of IL-8, without reducing that of GRO-, ICAM-1, IL-1 and IL-6. In conclusion, intracellular delivery of "decoy" molecules aimed to compete with the TF, NF-B, is a promising strategy to obtain inhibition of of IL-8 gene transcription.
