Inflammatory response of microglia to prions is controlled by sialylation of PrP(Sc)

Neuroinflammation is recognized as one of the obligatory pathogenic features of neurodegenerative diseases including Alzheimer’s, Parkinson’s or prion diseases. In prion diseases, space and time correlations between deposition of disease-associated, pathogenic form of the prion protein or PrP(Sc) and microglial-mediated neuroinflammation has been established. Yet, it remains unclear whether activation of microglia is triggered directly by a contact with PrP(Sc), and what molecular features of PrP(Sc) microglia sense and respond to that drive microglia to inflammatory states. The current study asked the questions whether PrP(Sc) can directly trigger activation of microglia and whether the degree of microglia response depends on the nature of terminal carbohydrate groups on the surface of PrP(Sc) particles. PrP(Sc) was purified from brains of mice infected with mouse-adapted prion strain 22L or neuroblastoma N2a cells stably infected with 22L. BV2 microglial cells or primary microglia were cultured in the presence of purified 22L. We found that exposure of BV2 cells or primary microglia to purified PrP(Sc) triggered proinflammatory responses characterized by an increase in the levels of TNFα, IL6, nitric oxide (NO) and expression of inducible Nitric Oxide Synthase (iNOS). Very similar patterns of inflammatory response were induced by PrP(Sc) purified from mouse brains and neuroblastoma cells arguing that microglia response is independent of the source of PrP(Sc). To test whether the microglial response is mediated by carbohydrate epitopes on PrP(Sc) surface, the levels of sialylation of PrP(Sc) N-linked glycans was altered by treatment of purified PrP(Sc) with neuraminidase. Partial cleavage of sialic acid residues was found to boost the inflammatory response of microglia to PrP(Sc). Moreover, transient degradation of Iκβα observed upon treatment with partially desialylated PrP(Sc) suggests that canonical NFκB activation pathway is involved in inflammatory response. The current study is the first to demonstrate that PrP(Sc) can directly trigger inflammatory response in microglia. In addition, this work provides direct evidence that the chemical nature of the carbohydrate groups on PrP(Sc) surface is important for microglial activation.