Antisense IRAK-2 Oligonucleotide Blocks Il-1-Stimulated NF-κB Activation and ICAM-1 Expression in Cultured Endothelial Cells

Phosphorothioate oligodeoxynucleotide (ODN) was designed antisense to sequences of the recently cloned human IL-1 receptor associated kinase-2 (IRAK-2). Antisense IRAK-2 ODN was delivered by lipofectin encapsulation into cultured endothelial cells. The levels of NF-κB, surface expression of intracellular adhesion molecule-1 (ICAM-1), ICAM-1 and IRAK-2 mRNAs were measured by sandwich ELISA, ELISA on cells in situ, and semiquantitative reverse transcription-PCR (RT-PCR), respectively. Antisense IRAK-2 ODN inhibited IL-1-induced NF-κB activation and surface expression of ICAM-1 in a concentration (1–4 μg)- and time (5–24 h)-dependent fashion. A maximum inhibition of NF-κB activation or surface expression of ICAM-l occurred when the cells were incubated with antisense IRAK-2 ODN 3 μg for 8 h. IL-1-induced ICAM-1 mRNA expression was also inhibited after treatment of cells with antisense IRAK-2 ODN 3 μg for 8 h. The attenuation of the cellular response to IL-1 caused by antisense IRAK-2 ODN correlated with a reduction of IRAK-2 expression. These data suggest that antisense IRAK-2 ODN may share a role in the design of antiinflammatory therapeutics.