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Neurological sequelae induced by alphavirus infection of the CNS are attenuated by treatment with the glutamine antagonist 6-diazo-5-oxo-l-norleucine

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Abstract

Recovery from encephalomyelitis induced by infection with mosquito-borne alphaviruses is associated with a high risk of lifelong debilitating neurological deficits. Infection of mice with the prototypic alphavirus, Sindbis virus, provides an animal model with which to study disease mechanisms and examine potential therapeutics. Infectious virus is cleared from the brain within a week after infection, but viral RNA is cleared slowly and persists for the life of the animal. However, no studies have examined the effect of infection on neurocognitive function over time. In the present study, we examined neurocognitive function at different phases of infection in 5-week-old C57BL/6 mice intranasally inoculated with Sindbis virus. At the peak of active virus infection, mice demonstrated hyperactivity, decreased anxiety, and marked hippocampal-dependent memory deficits, the latter of which persisted beyond clearance of infectious virus and resolution of clinical signs of disease. Previous studies indicate that neuronal damage during alphavirus encephalomyelitis is primarily due to inflammatory cell infiltration and glutamate excitotoxicity rather than directly by virus infection. Therefore, mice were treated with 6-diazo-5-oxo-l-norleucine (DON), a glutamine antagonist that can suppress both the immune response and excitotoxicity. Treatment with DON decreased inflammatory cell infiltration and cell death in the hippocampus and partially prevented development of clinical signs and neurocognitive impairment despite the presence of infectious virus and high viral RNA levels. This study presents the first report of neurocognitive sequelae in mice with alphavirus encephalomyelitis and provides a model system for further elucidation of the pathogenesis of virus infection and assessment of potential therapies.

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Acknowledgments

The following grants were used to fund this research: NIH grants R01 NS038932 (DEG), R01 NS087539 (DEG), T32 8T32OD011089 (VKB), P30 MH075673 (BSS), and R03 DA032470 (BSS), as well as a pilot grant from the Brain Science Institute of Johns Hopkins University School of Medicine. The funding sources played no role in the conduct of the research, preparation of the paper, or decision to submit the article for publication. The authors would like to thank Joseph Mankowski, Kelly Metcalf Pate, Lisa Mangus, and Claire Lyons along with the Retrovirus group at Johns Hopkins University for the use of and assistance with their microscope and imaging software. We would additionally like to express our appreciation to Sivabalan Manivannan for his assistance in preparing the DON stock solution. The authors declare no competing financial interests.

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    Authors and Affiliations

    1. Brain Science Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

      Michelle C. Potter, Robert W. Mathey, Jesse Alt, Camilo Rojas & Barbara S. Slusher

    2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

      Michelle C. Potter & Barbara S. Slusher

    3. Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

      Victoria K. Baxter & Camilo Rojas

    4. Department of Psychiatry, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

      Barbara S. Slusher

    5. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

      Barbara S. Slusher

    6. W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, 21205, USA

      Victoria K. Baxter & Diane E. Griffin

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    1. Michelle C. Potter
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    Michelle C. Potter and Victoria K. Baxter contributed equally to this work.

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    Fig. S1

    Gastrointestinal toxicity associated with high dose (0.6 mg/kg) DON treatment. Representative photomicrographs of hematoxylin & eosin-stained large intestine from (a) untreated, mock-infected and (b) high dose (0.6 mg/kg) DON, mock-infected mice at 7 DPI. High dose DON-treatment resulted in intestinal dilatation with loss of columnar epithelium and decreased cellularity (100× magnification; scale bar = 500 μm). (c) SINV-infected mice receiving high dose (0.6 mg/kg) DON regained body weight lost by 28 DPI (N = 3-4 mice per group) (JPEG 707 kb)

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    Potter, M.C., Baxter, V.K., Mathey, R.W. et al. Neurological sequelae induced by alphavirus infection of the CNS are attenuated by treatment with the glutamine antagonist 6-diazo-5-oxo-l-norleucine. J. Neurovirol. 21, 159–173 (2015). https://doi.org/10.1007/s13365-015-0314-6

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