IL-1R–Associated Kinase-1 Mediates Protein Kinase Cδ-Induced IL-1β Production in Monocytes

Abstract

The role of IL-1R-associated kinase (IRAK)1 and its interaction with protein kinase C (PKC)δ in monocytes to regulate IL-1β production has not been reported so far. The present study thus investigates such mechanisms in the THP1 cell line and human monocytes. PMA treatment to THP1 cells induced CD11b, TLR2, TLR4, CD36, IRAK1, IRAK3, and IRAK4 expression, IRAK1 kinase activity, PKCδ and JNK phosphorylation, AP-1 and NF-κB activation, and secretory IL-1β production. Moreover, PMA-induced IL-1β production was significantly reduced in the presence of TLR2, TLR4, and CD11b Abs. Rottlerin, a PKCδ-specific inhibitor, significantly reduced PMA-induced IL-1β production as well as CD11b, TLR2 expression, and IRAK1-JNK activation. In PKCδ wild-type overexpressing THP1 cells, IRAK1 kinase activity and IL-1β production were significantly augmented, whereas recombinant inactive PKCδ and PKCδ small interfering RNA significantly inhibited basal and PMA-induced IRAK1 activation and IL-1β production. Endogenous PKCδ-IRAK1 interaction was observed in quiescent cells, and this interaction was regulated by PMA. IRAK1/4 inhibitors, their small interfering RNAs, and JNK inhibitor also attenuated PMA-induced IL-1β production. NF-κB activation inhibitor and SN50 peptide inhibitor, however, failed to affect PMA-induced IL-1β production. A similar role of IRAK1 in IL-1β production and its regulation by PKCδ was evident in the primary human monocytes, thus signifying the importance of our finding. To our knowledge, the results obtained demonstrate for the first time that IRAK1 and PKCδ functionally interact to regulate IL-1β production in monocytic cells. A novel mechanism of IL-1β production that involves TLR2, CD11b, and the PKCδ/IRAK1/JNK/AP-1 axis is thus being proposed.