{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:06:43Z","timestamp":1788394003128,"version":"build-2803163510"},"reference-count":54,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,11,2]],"date-time":"2018-11-02T00:00:00Z","timestamp":1541116800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Venezuelan equine encephalitis virus (VEEV) is an arbovirus that is associated with robust inflammation that contributes to neurodegenerative phenotypes. In addition to triggering central nervous system (CNS) inflammation, VEEV will also induce mitochondrial dysfunction, resulting in increased cellular apoptosis. In this study, we utilize the TC-83 strain of VEEV to determine the role of mitochondrial oxidative stress in mediating inflammation elicited by murine brain microglial cells. Using an in vitro model, we show that murine microglia are susceptible to TC-83 infection, and that these cells undergo mitochondrial stress as the result of infection. We also indicate that bystander microglia contribute more significantly to the overall inflammatory load than directly infected microglia. Use of a mitochondrial targeted antioxidant, mitoquinone mesylate, greatly reduced the pro-inflammatory cytokines released by both direct infected and bystander microglia. Our data suggest that release of interleukin-1\u03b2, a key instigator of neuroinflammation during VEEV infection, may be the direct result of accumulating mitochondrial stress. This data improves our understanding inflammation elicited by murine microglia and will aid in the development of more accurate in vitro and in vivo murine model of VEEV-induced neuroinflammation.<\/jats:p>","DOI":"10.3390\/v10110606","type":"journal-article","created":{"date-parts":[[2018,11,5]],"date-time":"2018-11-05T04:26:39Z","timestamp":1541391999000},"page":"606","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Mitochondrial-Directed Antioxidant Reduces Microglial-Induced Inflammation in Murine In Vitro Model of TC-83 Infection"],"prefix":"10.3390","volume":"10","author":[{"given":"Forrest","family":"Keck","sequence":"first","affiliation":[{"name":"National Center for Biodefense and Infectious Diseases, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daud","family":"Khan","sequence":"additional","affiliation":[{"name":"Krasnow Institute for Advanced Study, George Mason University, Fairfax, VA 22030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Roberts","sequence":"additional","affiliation":[{"name":"Leidos Health Sciences, 5202 Presidents Court, Suite 110, Frederick, MD 21704, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nitin","family":"Agrawal","sequence":"additional","affiliation":[{"name":"Krasnow Institute for Advanced Study, George Mason University, Fairfax, VA 22030, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nishank","family":"Bhalla","sequence":"additional","affiliation":[{"name":"National Center for Biodefense and Infectious Diseases, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aarthi","family":"Narayanan","sequence":"additional","affiliation":[{"name":"National Center for Biodefense and Infectious Diseases, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,11,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"721","DOI":"10.2217\/fvl.11.50","article-title":"Endemic Venezuelan equine encephalitis in the Americas: Hidden under the dengue umbrella","volume":"6","author":"Aguilar","year":"2011","journal-title":"Future Virol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1146\/annurev.micro.55.1.235","article-title":"Biological Weapons\u2014A Primer for Microbiologists","volume":"55","author":"Hawley","year":"2001","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1146\/annurev.ento.49.061802.123422","article-title":"Venezuelan Equine Encephalitis","volume":"49","author":"Weaver","year":"2004","journal-title":"Annu. Rev. Entomol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"D80","DOI":"10.1016\/j.vaccine.2009.07.095","article-title":"Vaccines for Venezuelan equine encephalitis","volume":"27","author":"Paessler","year":"2009","journal-title":"Vaccine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1128\/mr.58.3.491-562.1994","article-title":"The alphaviruses: Gene expression, replication, and evolution","volume":"58","author":"Strauss","year":"1994","journal-title":"Microbiol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2243","DOI":"10.1007\/BF01323243","article-title":"Spread of Venezuelan equine encephalitis virus in mice olfactory tract","volume":"140","author":"Ryzhikov","year":"1995","journal-title":"Arch. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1006\/viro.2001.0878","article-title":"Immunopathogenesis and Immune Modulation of Venezuelan Equine Encephalitis Virus-Induced Disease in the Mouse","volume":"284","author":"Charles","year":"2001","journal-title":"Virology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1021\/acschemneuro.6b00043","article-title":"The Olfactory Bulb: An Immunosensory Effector Organ during Neurotropic Viral Infections","volume":"7","author":"Durrant","year":"2016","journal-title":"ACS Chem. Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1177\/030098589102800509","article-title":"Pathogenesis of Venezuelan Equine Encephalitis Virus Infection in Mice and Hamsters","volume":"28","author":"Jackson","year":"1991","journal-title":"Vet. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1007\/s004010050626","article-title":"Apoptotic cell death is an important cause of neuronal injury in experimental Venezuelan equine encephalitis virus infection of mice","volume":"93","author":"Jackson","year":"1997","journal-title":"Acta Neuropathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/S0165-5728(00)00290-3","article-title":"Inflammation is a component of neurodegeneration in response to Venezuelan equine encephalitis virus infection in mice","volume":"109","author":"Schoneboom","year":"2000","journal-title":"J. Neuroimmunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"342","DOI":"10.3109\/13550289909029475","article-title":"Astrocytes as targets for Venezuelan equine encephalitis virus infection","volume":"5","author":"Schoneboom","year":"1999","journal-title":"J. Neurovirol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1836","DOI":"10.1099\/vir.0.010280-0","article-title":"Oligonucleotide array analysis of Toll-like receptors and associated signalling genes in Venezuelan equine encephalitis virus-infected mouse brain","volume":"90","author":"Sharma","year":"2009","journal-title":"J. Gen. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kehn-Hall, K., Narayanan, A., Lundberg, L., Sampey, G., Pinkham, C., Guendel, I., Duyne, R.V., Senina, S., Schultz, K.L., and Stavale, E. (2012). Modulation of GSK-3\u03b2 Activity in Venezuelan Equine Encephalitis Virus Infection. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0034761"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1089\/107999000312621","article-title":"Early expression of IFN-alpha\/beta and iNOS in the brains of Venezuelan equine encephalitis virus-infected mice","volume":"20","author":"Schoneboom","year":"2000","journal-title":"J. Interferon Cytokine Res. Off. J. Int. Soc. Interferon Cytokine Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/j.vetmic.2012.11.021","article-title":"Production of IL-8, IL-17, IFN-gamma and IP-10 in human astrocytes correlates with alphavirus attenuation","volume":"163","author":"Peng","year":"2013","journal-title":"Vet. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1111\/jnc.13667","article-title":"Targeting innate immunity for neurodegenerative disorders of the central nervous system","volume":"138","author":"Andreasson","year":"2016","journal-title":"J. Neurochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"114393","DOI":"10.18632\/oncotarget.23106","article-title":"Microglia at center stage: A comprehensive review about the versatile and unique residential macrophages of the central nervous system","volume":"8","author":"Lannes","year":"2017","journal-title":"Oncotarget"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Ransohoff, R.M., and Khoury, J.E. (2016). Microglia in Health and Disease. Cold Spring Harb. Perspect. Biol., 8.","DOI":"10.1101\/cshperspect.a020560"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Sanchez-Guajardo, V., Barnum, C.J., Tansey, M.G., and Romero-Ramos, M. (2013). Neuroimmunological processes in Parkinson\u2019s disease and their relation to \u03b1-synuclein: Microglia as the referee between neuronal processes and peripheral immunity. ASN Neuro, 5.","DOI":"10.1042\/AN20120066"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/S0304-3940(00)00977-0","article-title":"Temporal profile of release of interleukin-1\u03b2 in neurotrauma","volume":"284","author":"Fassbender","year":"2000","journal-title":"Neurosci. Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1002\/glia.10161","article-title":"Microglia as a source and target of cytokines","volume":"40","author":"Hanisch","year":"2002","journal-title":"Glia"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"457","DOI":"10.3109\/08830189809043005","article-title":"Interleukin-1, Interleukin-1 Receptors and Interleukin-1 Receptor Antagonist","volume":"16","author":"Dinarello","year":"1998","journal-title":"Int. Rev. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1074\/jbc.M406968200","article-title":"Distinct Roles for the \u03b1 and \u03b2 Subunits in the Functions of Integrin \u03b1M\u03b22","volume":"280","author":"Solovjov","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2840","DOI":"10.1128\/JVI.74.6.2840-2846.2000","article-title":"Hepatitis B Virus X Protein Colocalizes to Mitochondria with a Human Voltage-Dependent Anion Channel, HVDAC3, and Alters Its Transmembrane Potential","volume":"74","author":"Rahmani","year":"2000","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7721","DOI":"10.1128\/MCB.21.22.7721-7730.2001","article-title":"Mitochondrially Associated Hepatitis B Virus X Protein Constitutively Activates Transcription Factors STAT-3 and NF-\u03baB via Oxidative Stress","volume":"21","author":"Waris","year":"2001","journal-title":"Mol. Cell. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Kim, S.-J., Khan, M., Quan, J., Till, A., Subramani, S., and Siddiqui, A. (2013). Hepatitis B Virus Disrupts Mitochondrial Dynamics: Induces Fission and Mitophagy to Attenuate Apoptosis. PLOS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003722"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6413","DOI":"10.1073\/pnas.1321114111","article-title":"Hepatitis C virus triggers mitochondrial fission and attenuates apoptosis to promote viral persistence","volume":"111","author":"Kim","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2376","DOI":"10.1126\/science.294.5550.2376","article-title":"Calcium Signaling by HBx Protein in Hepatitis B Virus DNA Replication","volume":"294","author":"Bouchard","year":"2001","journal-title":"Science"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1016\/j.virol.2013.11.023","article-title":"Reactive oxygen species activate NF\u03baB (p65) and p53 and induce apoptosis in RVFV infected liver cells","volume":"449","author":"Narayanan","year":"2014","journal-title":"Virology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1128\/JVI.02192-12","article-title":"The C-Terminal Region of Rift Valley Fever Virus NSm Protein Targets the Protein to the Mitochondrial Outer Membrane and Exerts Antiapoptotic Function","volume":"87","author":"Terasaki","year":"2013","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1849","DOI":"10.1080\/21505594.2016.1276690","article-title":"Altered mitochondrial dynamics as a consequence of Venezuelan Equine encephalitis virus infection","volume":"8","author":"Keck","year":"2017","journal-title":"Virulence"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Keck, F., Kortchak, S., Bakovic, A., Roberts, B., Agrawal, N., and Narayanan, A. (2018). Direct and Indirect Pro-Inflammatory Cytokine Response Resulting from TC-83 Infection of Glial Cells. Virulence.","DOI":"10.1080\/21505594.2018.1509668"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1186\/1743-422X-8-197","article-title":"Role of adhesion molecules and inflammation in Venezuelan equine encephalitis virus infected mouse brain","volume":"8","author":"Sharma","year":"2011","journal-title":"Virol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.bbrc.2010.03.091","article-title":"Analysis of microRNAs induced by Venezuelan equine encephalitis virus infection in mouse brain","volume":"395","author":"Bhomia","year":"2010","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Gupta, P., Sharma, A., Han, J., Yang, A., Bhomia, M., Knollmann-Ritschel, B., Puri, R.K., and Maheshwari, R.K. (2017). Differential host gene responses from infection with neurovirulent and partially-neurovirulent strains of Venezuelan equine encephalitis virus. BMC Infect. Dis., 17.","DOI":"10.1186\/s12879-017-2355-3"},{"key":"ref_37","first-page":"401","article-title":"Mitochondrial distribution and function in herpes simplex virus-infected cells","volume":"81","author":"Murata","year":"2000","journal-title":"J. Gen. Virol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ambrosius, B., Faissner, S., Guse, K., von Lehe, M., Grunwald, T., Gold, R., Grewe, B., and Chan, A. (2017). Teriflunomide and monomethylfumarate target HIV-induced neuroinflammation and neurotoxicity. J. Neuroinflamm., 14.","DOI":"10.1186\/s12974-017-0829-2"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1170","DOI":"10.1016\/j.imbio.2015.06.002","article-title":"Novel evidence of microglial immune response in impairment of Dengue infection of CNS","volume":"220","author":"Bhatt","year":"2015","journal-title":"Immunobiology"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Das, S., Dutta, K., Kumawat, K.L., Ghoshal, A., Adhya, D., and Basu, A. (2011). Abrogated Inflammatory Response Promotes Neurogenesis in a Murine Model of Japanese Encephalitis. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0017225"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1111\/jnc.12933","article-title":"HIV-1 Tat C modulates NOX2 and NOX4 expressions through miR-17 in a human microglial cell line","volume":"131","author":"Jadhav","year":"2014","journal-title":"J. Neurochem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"787023","DOI":"10.1155\/2014\/787023","article-title":"Roles of TLR3 and RIG-I in mediating the inflammatory response in mouse microglia following Japanese encephalitis virus infection","volume":"2014","author":"Jiang","year":"2014","journal-title":"J. Immunol. Res."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Lundberg, L., Carey, B., and Kehn-Hall, K. (2017). Venezuelan Equine Encephalitis Virus Capsid\u2014The Clever Caper. Viruses, 9.","DOI":"10.3389\/fmicb.2018.03126"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Amaya, M., Voss, K., Sampey, G., Senina, S., de la Fuente, C., Mueller, C., Calvert, V., Kehn-Hall, K., Carpenter, C., and Kashanchi, F. (2014). The role of IKK\u03b2 in Venezuelan equine encephalitis virus infection. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0086745"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1111\/j.1574-695X.2007.00319.x","article-title":"Gene expression profiling of nonhuman primates exposed to aerosolized Venezuelan equine encephalitis virus","volume":"51","author":"Koterski","year":"2007","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1007\/BF01317831","article-title":"Interferon induction and sensitivity as correlates to virulence of Venezuelan encephalitis viruses for hamsters","volume":"51","author":"Jahrling","year":"1976","journal-title":"Arch. Virol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"762","DOI":"10.4269\/ajtmh.1967.16.762","article-title":"Live, Attenuated Venezuelan Equine Encephalomyelitis Virus Vaccine","volume":"16","author":"Alevizatos","year":"1967","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1719","DOI":"10.1111\/bph.13426","article-title":"Microglia antioxidant systems and redox signalling","volume":"174","author":"Vilhardt","year":"2017","journal-title":"Br. J. Pharmacol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"686","DOI":"10.1016\/j.freeradbiomed.2008.05.026","article-title":"Reactive oxygen species up-regulate CD11b in microglia via nitric oxide: Implications for neurodegenerative diseases","volume":"45","author":"Roy","year":"2008","journal-title":"Free Radic. Biol. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"14971","DOI":"10.1074\/jbc.M600236200","article-title":"Up-regulation of Microglial CD11b Expression by Nitric Oxide","volume":"281","author":"Roy","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Rajalakshmy, A.R., Malathi, J., and Madhavan, H.N. (2015). Hepatitis C Virus NS3 Mediated Microglial Inflammation via TLR2\/TLR6 MyD88\/NF-\u03baB Pathway and Toll Like Receptor Ligand Treatment Furnished Immune Tolerance. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0125419"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1186\/1742-2094-8-123","article-title":"Reactive oxygen species drive herpes simplex virus (HSV)-1-induced proinflammatory cytokine production by murine microglia","volume":"8","author":"Hu","year":"2011","journal-title":"J. Neuroinflamm."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.jneuroim.2013.12.002","article-title":"Role of peripheral immune response in microglia activation and regulation of brain chemokine and proinflammatory cytokine responses induced during VSV encephalitis","volume":"267","author":"Steel","year":"2014","journal-title":"J. Neuroimmunol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1007\/s13365-013-0188-4","article-title":"Microglia-mediated neuroinflammation is an amplifier of virus-induced neuropathology","volume":"20","year":"2014","journal-title":"J. Neurovirol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/11\/606\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:27:35Z","timestamp":1760196455000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/10\/11\/606"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,2]]},"references-count":54,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2018,11]]}},"alternative-id":["v10110606"],"URL":"https:\/\/doi.org\/10.3390\/v10110606","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,2]]}}}