Type-1 interferons contribute to oxygen glucose deprivation induced neuro-inflammation in BE(2)M17 human neuroblastoma cells

• Published in: Journal of neuroinflammation 2014-03, Vol.11 (1), p.43-43, Article 43
• Main Authors: Minter, Myles Robert, Zhang, Moses, Ates, Robert Charles, Taylor, Juliet Marie, Crack, Peter John
• Format: Article
• Language: English
• Subjects: Analysis of Variance; Autoimmune diseases; Brain damage; Brain research; Cell Line, Tumor; Chemokines; Cytokines; Cytokines - genetics; Cytokines - metabolism; Gene Expression Regulation - physiology; Glucose; Glucose - deficiency; Humans; Hypoxia; Hypoxia - physiopathology; Immune system; Inflammation - metabolism; Injuries; Ligands; Nervous system; Neuroblastoma - pathology; Neurons; Oxygen; Phosphorylation; Protein Isoforms; Receptor, Interferon alpha-beta - genetics; Receptor, Interferon alpha-beta - metabolism; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Rodents; Signal Transduction - genetics; Signal Transduction - physiology; STAT1 Transcription Factor - metabolism; Transfection
• ISSN: 1742-2094; 1742-2094
• DOI: 10.1186/1742-2094-11-43

Hypoxic-ischaemic injuries such as stroke and traumatic brain injury exhibit features of a distinct neuro-inflammatory response in the hours and days post-injury. Microglial activation, elevated pro-inflammatory cytokines and macrophage infiltration contribute to core tissue damage and contribute to secondary injury within a region termed the penumbra. Type-1 interferons (IFNs) are a super-family of pleiotropic cytokines that regulate pro-inflammatory gene transcription via the classical Jak/Stat pathway; however their role in hypoxia-ischaemia and central nervous system neuro-inflammation remains unknown. Using an in vitro approach, this study investigated the role of type-1 IFN signalling in an inflammatory setting induced by oxygen glucose deprivation (OGD). Human BE(2)M17 neuroblastoma cells or cells expressing a type-1 interferon-α receptor 1 (IFNAR1) shRNA or negative control shRNA knockdown construct were subjected to 4.5 h OGD and a time-course reperfusion period (0 to 24 h). Q-PCR was used to evaluate IFNα, IFNβ, IL-1β, IL-6 and TNF-α cytokine expression levels. Phosphorylation of signal transducers and activators of transcription (STAT)-1, STAT-3 and cleavage of caspase-3 was detected by western blot analysis. Post-OGD cellular viability was measured using a MTT assay. Elevated IFNα and IFNβ expression was detected during reperfusion post-OGD in parental M17 cells. This correlated with enhanced phosphorylation of STAT-1, a downstream type-1 IFN signalling mediator. Significantly, ablation of type-1 IFN signalling, through IFNAR1 knockdown, reduced IFNα, IFNβ, IL-6 and TNF-α expression in response to OGD. In addition, MTT assay confirmed the IFNAR1 knockdown cells were protected against OGD compared to negative control cells with reduced pro-apoptotic cleaved caspase-3 levels. This study confirms a role for type-1 IFN signalling in the neuro-inflammatory response following OGD in vitro and suggests its modulation through therapeutic blockade of IFNAR1 may be beneficial in reducing hypoxia-induced neuro-inflammation.