Attenuation of interleukin-8 production by inhibiting nuclear factor-kappaB translocation using decoy oligonucleotides

Interleukin-8 (IL-8), a monocyte-derived neutrophil chemoattractant factor, is a polymorphonuclear neutrophil chemotaxin that is involved in a number of inflammatory disorders. Transcription of the IL-8 gene is controlled by regulatory proteins, including nuclear factor-kappa B (NF kappa B), a famil y of proteins that is important in the transcriptional control of a number of genes. When cells are activated, NF-kappa B translocates from the cytopl asm to the nucleus, where it activates transcription by binding to a specif ic sequence within the 5' untranslated region of the gene. During transloca tion, NF-kappa B is potentially susceptible to diversion by oligonucleotide s that contain the binding sequence for this protein. In the current study, we produced phosphorothioate-modified oligonucleotides containing the spec ific DNA sequence that NF-kappa B binds within the IL-8 gene. We then inves tigated the effects of transfection of monocytes with these oligonucleotide s on interleukin-1 beta (IL-1 beta)-stimulated IL-8 production IL-8 mRNA ex pression, and NF-kappa B binding activity. We found that transfection with these oligonucleotides significantly inhibited monocyte IL-8 production. A single-stranded oligonucleotide with two copies of the NF-kappa B-binding s equence was the most potent of those tested. This single-stranded oligonucl eotide also inhibited IL-1 beta-induced translocation of NF-kappa B to the nucleus and reduced IL-8 mRNA expression. These studies demonstrated that m onocyte production of IL-8 can be attenuated using a single-stranded oligon ucleotide that binds a transcriptional activating protein before it translo cates to the cell nucleus. This approach ultimately may be useful in the co ntrol of inflammation involved in a number of diseases. (C) 2000 Elsevier S cience Inc.