It was already known that caspase was necessary for the activation of T cells after recognition of Borrelia spp. by PRR 26, which is in line with our results. The induction of pro-inflammatory cytokines IL-1β and IL-17 by Borrelia was

caspase-1 dependent, and both cytokines have been shown already to play a role in the pathogenesis caused by Borrelia 27–29. In line with this, we have demonstrated that stimulation of macrophages and spleen cells by Borrelia resulted in production of IL-1β, IL-6, IL-17 and IFN-γ (Fig. 1). In addition, after intra-articular (i.a.) injection with Borrelia we observed less cell influx and cytokine production in caspase-1-deficient animals as compared to the WT animals (Fig. 3). We observed differences in IL-6 production after Borrelia stimulation between caspase-1-deficient peritoneal macrophages and PMN isolated from the knee of caspase-1 knockout animals. This difference can be explained LY2606368 by the fact that different types of cells are involved and different time points were used in these assays. In the patella washouts assays, the main cell types that could LBH589 produce IL-6 are granulocytes (PMN) and synovial fibroblasts. These cells may respond differently after exposure to Borrelia when compared

to peritoneal macrophages. The other explanation could be that the synovial cells were only 4 h exposed to Borrelia whereas the peritoneal macrophages were treated for 24 h with Borrelia. We also describe that Borrelia-induced IL-1β is the Flavopiridol (Alvocidib) main inducer of IL-17 production after stimulation

with Borrelia (Fig. 4). Furthermore, caspase-1-cleaved IL-18 is responsible for induction of IFN-γ by Borrelia spp. (Fig. 5A). Caspase-1 is crucial for Borrelia-induced IFN-γ production, as caspase-1-deficient mice produced almost no IFN-γ. The exact role of IFN-γ in the host defense against Borrelia has not yet been elucidated. On the one hand, the induction of Borrelia-induced arthritis does not seem to be dependent on IFN-γ 30–32, and it has been reported that mice with a disrupted IFN-γ gene are more susceptible to autoimmune disorders such as EAE and collagen-induced arthritis 33, 34. On the other hand, several groups have proposed a role for IFN-γ-producing T cells in Lyme arthritis 34, 35. In patients infected with Borrelia, high levels of IFN-γ were measured 36. In line with this, we found that IFN-γ is produced in large amounts by spleen cells after stimulation with Borrelia spirochetes. Dame et al. 37 described that IFN-γ in combination with B. burgdorferi cooperatively induced upregulation of endothelial cell genes, causing more T-cell infiltration. It has been known that IFN-γ modulates other T-cell cytokines. It has been described before that IFN-γ controls or modulates Th17 responses 38, 39, but until now this has not been demonstrated for Borrelia-induced Th17 responses.