Indeed, treatment with rhIL-10 significantly reduced both CD8+ and CD4+ T-cell proliferation (Fig. 7C), thus proving a central role of IL-10 in the regulation of the T-cell response to allogenic monocytes. In this study, we demonstrated the role of the IRAK4 kinase as a differential switch between TLR-induced pro-inflammatory and anti-inflammatory cytokine production. This observation is of interest as to date IRAK4 is mainly being viewed as a central executor of the MyD88 pathway that unselectively transduces all signals downstream of MyD88. As previously described in IRAK4-deficient mice [26], stimulation of IRAK4 knockdown monocytes with TLR4 and TLR2 ligands resulted in markedly reduced pro-inflammatory cytokine

secretion such as TNF and IL-12 (Fig. 2A–C) and selleck screening library concomitant reduction of the NF-κB subunits p50 and p65 responsible for the transcription of these cytokines (Fig. 2D). The results obtained are further in accordance Bortezomib price with observations made in cells of IRAK4- and MyD88-deficient patients. There, TLR stimulation fails to activate the NF-κB pathway and cells display an impaired cytokine response after stimulation with agonists for TLR-1, TLR-2, TLR-4, TLR-5, TLR-7, and TLR-8 [17-20]. Similarly, siRNA-mediated silencing of MyD88 caused a significant reduction in TNF and IL-12 cytokine production in human monocytes (Fig. 4C,D), highlighting the cooperative interaction

of MyD88 and IRAK4 in TLR-induced synthesis of pro-inflammatory cytokines such as TNF and IL-12. IRAK4-deficient patients are more susceptible to infections with pyogenic bacteria, especially Gram-positive species such as S. pneumoniae and/or S. aureus [18]. Consistently, the predisposition to S. aureus infections was also reported in IRAK4-deficient mice [26]. Until now, little is known about the role of monocytes in response to these pathogens albeit these cells belong to those first to encounter bacterial pathogens in blood stream infections.

In this study, we tested the role of IRAK4 in the TLR-mediated response of human monocytes to bacterial infections. In particular, our results showed diminished IL-12 heptaminol secretion and elevated TNF and IL-10 levels after treatment with live S. aureus and S. pneumoniae (Fig. 1C). To further assess these findings we conducted MyD88 knockdown experiments, obtaining concomitantly reduced IL-12 and IL-10 secretion. However, TNF levels were only slightly diminished (Fig. 4E). This strongly suggests that IL-12 and IL-10 secretion evoked by bacterial infections is MyD88-dependent, whereas TNF production is also regulated in a MyD88-independent fashion, possibly triggered via TRIF or cytosolic PRR-induced IFN-I [27]. In general, bacteria represent complexes of multiple ligands that can be sensed by membrane-bound as well as cytosolic PRR. The most prominent difference between stimulation with bacteria and single TLR ligands was an increase in TNF release under IRAK4-silencing conditions (Fig. 1C) and under rapamycin treatment (Fig. 5B).