A role for DNA-dependent activator of interferon regulatory factor in the recognition of herpes simplex virus type 1 by glial cells

• Published in: Journal of neuroinflammation Vol. 8; no. 1; p. 99
• Main Authors: Furr, Samantha R, Chauhan, Vinita S, Moerdyk-Schauwecker, Megan J, Marriott, Ian
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 12-08-2011; BioMed Central; BMC
• Subjects: Animals; astrocytes; Astrocytes - cytology; Astrocytes - metabolism; Astrocytes - virology; Cell Death; Cells, Cultured; DNA polymerases; DNA, B-Form - chemistry; DNA, B-Form - metabolism; DNA-dependent activator of IFN regulatory factor; encephalitis; Encephalitis, Viral - immunology; Encephalitis, Viral - physiopathology; Encephalitis, Viral - virology; Female; Herpes simplex virus; herpes simplex virus-1; Herpesvirus 1, Human - immunology; Humans; Immunity, Innate - immunology; innate immunity; Interferon Regulatory Factors - genetics; Interferon Regulatory Factors - metabolism; Interleukin-6 - immunology; Mice; Mice, Inbred C57BL; microglia; Microglia - cytology; Microglia - metabolism; Microglia - virology; Neuroglia; neuroinflammation; Neurons - pathology; Physiological aspects; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; Tumor Necrosis Factor-alpha - immunology; United States
• ISSN: 1742-2094; 1742-2094
• DOI: 10.1186/1742-2094-8-99

The rapid onset of potentially lethal neuroinflammation is a defining feature of viral encephalitis. Microglia and astrocytes are likely to play a significant role in viral encephalitis pathophysiology as they are ideally positioned to respond to invading central nervous system (CNS) pathogens by producing key inflammatory mediators. Recently, DNA-dependent activator of IFN regulatory factor (DAI) has been reported to function as an intracellular sensor for DNA viruses. To date, the expression and functional role of DAI in the inflammatory responses of resident CNS cells to neurotropic DNA viruses has not been reported.
Expression of DAI and its downstream effector molecules was determined in C57BL/6-derived microglia and astrocytes, either at rest or following exposure to herpes simplex virus type 1 (HSV-1) and/or murine gammaherpesvirus-68 (MHV-68), by immunoblot analysis. In addition, such expression was studied in ex vivo microglia/macrophages and astrocytes from uninfected animals or mice infected with HSV-1. Inflammatory cytokine production by glial cultures following transfection with a DAI specific ligand (B-DNA), or following HSV-1 challenge in the absence or presence of siRNA directed against DAI, was assessed by specific capture ELISA. The production of soluble neurotoxic mediators by HSV-1 infected glia following DAI knockdown was assessed by analysis of the susceptibility of neuron-like cells to conditioned glial media.
We show that isolated microglia and astrocytes constitutively express DAI and its effector molecules, and show that such expression is upregulated following DNA virus challenge. We demonstrate that these resident CNS cells express DAI in situ, and show that its expression is similarly elevated in a murine model of HSV-1 encephalitis. Importantly, we show B-DNA transfection can elicit inflammatory cytokine production by isolated glial cells and DAI knockdown can significantly reduce microglial and astrocyte responses to HSV-1. Finally, we demonstrate that HSV-1 challenged microglia and astrocytes release neurotoxic mediators and show that such production is significantly attenuated following DAI knockdown.
The functional expression of DAI by microglia and astrocytes may represent an important innate immune mechanism underlying the rapid and potentially lethal inflammation associated with neurotropic DNA virus infection.