Abstract
Inflammation in the absence of infection (sterile inflammation) contributes to acute injury and chronic disease. Cerebral ischemia is a devastating condition in which the primary injury is caused by reduced blood supply and is therefore sterile. The cytokine interleukin-1β (IL-1β) is a key contributor to ischemic brain injury and central inflammatory responses. The release of IL-1β is regulated by the protease caspase-1, and its activating complex, the inflammasome. Of the known inflammasomes the best characterized, and one that is perceived to sense sterile injury is formed by a pattern recognition receptor called NOD-like receptor pyrin domain containing three (NLRP3). A key feature of NLRP3-inflammasome dependent responses in vitro in macrophages is the requirement of an initial priming stimulus by a pathogen (PAMP), or damage associated molecular pattern (DAMP) respectively. We sought to determine the inflammatory responses of NLRP3-activating DAMPs on brain derived mixed glial cells in the absence of an initial priming stimulus in vitro. In cultured mouse mixed glia the DAMPs ATP, monosodium urate, and calcium pyrophosphate dehydrate crystals had no effect on the expression of IL-1α or IL-1β and induced release only when the cells were primed with a PAMP. In the absence of priming, these DAMPs did however induce inflammation via the production of IL-6 and CXCL1, and the release of the lysosomal protease cathepsin B. Furthermore, the acute phase protein serum amyloid A (SAA) acted as a priming stimulus on glial cells resulting in levels of IL-1 expression comparable to those induced by the PAMP lipopolysaccharide. In vivo, after cerebral ischemia, IL-1 production contributed to increased IL-6 and CXCL1 since these cytokines were profoundly reduced in the ischemic hemispheres from IL-1α/β double KO mice, although injury-induced cytokine responses were not abolished. Thus, DAMPs augment brain inflammation by directly stimulating production of glial derived inflammatory mediators. This is markedly enhanced by DAMP-induced IL-1-release-dependent responses that require a sterile endogenous priming stimulus such as SAA.
Highlights
Interleukin-1β (IL-1β) is a key pro-inflammatory cytokine that is central to the damaging inflammatory processes that accompany sterile disease (Dinarello, 2011)
We initially investigated the effects of these damage associated molecular pattern (DAMP) and LPS on the expression of genes typically associated with the inflammasome and priming e.g., IL-1β (Figure 1Ai), IL-1α (Figure 1Bi), caspase1 (Figure 1Ci), NLRP3 (Figure 1Di), and ASC (Figure 1Ei)
DAMPs had no effect on the expression of these genes except for calcium pyrophosphate dehydrate (CPPD) crystals, where a significant increase in the expression of IL-1β and IL-1α was observed (Figures 1Ai,Bi)
Summary
Interleukin-1β (IL-1β) is a key pro-inflammatory cytokine that is central to the damaging inflammatory processes that accompany sterile disease (Dinarello, 2011) This is true after an acute brain injury such as cerebral ischemia, or stroke, where IL1β is established as a major contributor to damage (Brough et al, 2011). It is produced during disease or after an injury as an inactive precursor (pro-IL-1β) by cells of the innate immune system such as macrophages, or in diseases of the central nervous system (CNS), by microglia (Denes et al, 2008). NLRP3 can be activated by a diverse array of disease associated molecules, where it oligomerizes with the adaptor ASC (apoptosis-associated speck-like protein containing a caspase recruitment domain) and caspase-1 to form the NLRP3inflammasome, resulting in the processing of pro- to mature IL-1β and its release
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