Journal of NeuroVirology

, Volume 17, Issue 3, pp 201–211 | Cite as

Cytomegalovirus-induced sensorineural hearing loss with persistent cochlear inflammation in neonatal mice

  • Scott J. Schachtele
  • Manohar B. Mutnal
  • Mark R. Schleiss
  • James R. LokensgardEmail author


Congenital cytomegalovirus (CMV) infection is the leading cause of sensorineural hearing loss (SNHL) in children. During murine (M)CMV-induced encephalitis, the immune response is important for both the control of viral dissemination and the clearance of virus from the brain. While the importance of CMV-induced SNHL has been described, the mechanisms surrounding its pathogenesis and the role of inflammatory responses remain unclear. This study presents a neonatal mouse model of profound SNHL in which MCMV preferentially infected both cochlear perilymphatic epithelial cells and spiral ganglion neurons. Interestingly, MCMV infection induced cochlear hair cell death by 21 days post-infection, despite a clear lack of direct infection of hair cells and the complete clearance of the virus from the cochlea by 14 dpi. Flow cytometric, immunohistochemical, and quantitative PCR analysis of MCMV-infected cochlea revealed a robust and chronic inflammatory response, including a prolonged increase in reactive oxygen species production by infiltrating macrophages. These data support a pivotal role for inflammation during MCMV-induced SNHL.


Cytomegalovirus SNHL Hearing Cochlea Inflammation Reactive oxygen species 



We thank Dr. Steven Juhn and Dr. Phil Peterson for their expertise and thoughtful input. This project was supported by Award Number R01 NS-038836 from the National Institute of Neurological Disorders and Stroke, as well as a PharmacoNeuroImmunology training grant (T32DA007097) funded by the National Institute on Drug Abuse. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NINDS, NIDA, or the NIH.


  1. Alam SA, Robinson BK, Huang J, Green SH (2007) Prosurvival and proapoptotic intracellular signaling in rat spiral ganglion neurons in vivo after the loss of hair cells. J Comp Neurol 503:832–852PubMedCrossRefGoogle Scholar
  2. Aminpour S, Tinling SP, Brodie HA (2005) Role of tumor necrosis factor-alpha in sensorineural hearing loss after bacterial meningitis. Otol Neurotol 26:602–609PubMedCrossRefGoogle Scholar
  3. Armien AG, Hu S, Little MR et al (2009) Chronic cortical and subcortical pathology with associated neurological deficits ensuing experimental herpes encephalitis. Brain Pathol 20(4):738–750PubMedCrossRefGoogle Scholar
  4. Bantug GR, Cekinovic D, Bradford R, Koontz T, Jonjic S, Britt WJ (2008) CD8+ T lymphocytes control murine cytomegalovirus replication in the central nervous system of newborn animals. J Immunol 181:2111–2123PubMedGoogle Scholar
  5. Caird DM, Klinke R (1987) The effect of inferior colliculus lesions on auditory evoked potentials. Electroencephalogr Clin Neurophysiol 68:237–240PubMedCrossRefGoogle Scholar
  6. Cekinovic D, Golemac M, Pugel EP, Tomac J, Cicin-Sain L, Slavuljica I, Bradford R, Misch S, Winkler TH, Mach M, Britt WJ, Jonjic S (2008) Passive immunization reduces murine cytomegalovirus-induced brain pathology in newborn mice. J Virol 82:12172–12180.PubMedCrossRefGoogle Scholar
  7. Cheeran MC, Hu S, Yager SL, Gekker G, Peterson PK, Lokensgard JR (2001) Cytomegalovirus induces cytokine and chemokine production differentially in microglia and astrocytes: antiviral implications. J Neurovirol 7:135–147PubMedCrossRefGoogle Scholar
  8. Cheeran MC, Gekker G, Hu S, Min X, Cox D, Lokensgard JR (2004) Intracerebral infection with murine cytomegalovirus induces CXCL10 and is restricted by adoptive transfer of splenocytes. J Neurovirol 10:152–162PubMedCrossRefGoogle Scholar
  9. Cheeran MC, Lokensgard JR, Schleiss MR (2009a) Neuropathogenesis of congenital cytomegalovirus infection: disease mechanisms and prospects for intervention. Clin Microbiol Rev 22:99–126, Table of ContentsPubMedCrossRefGoogle Scholar
  10. Cheeran MC, Mutnal MB, Hu S, Armien A, Lokensgard JR (2009b) Reduced lymphocyte infiltration during cytomegalovirus brain infection of interleukin-10-deficient mice. J Neurovirol 15:334–342PubMedCrossRefGoogle Scholar
  11. Chen TJ, Chen SS (1991) Generator study of brainstem auditory evoked potentials by a radiofrequency lesion method in rats. Exp Brain Res 85:537–542PubMedCrossRefGoogle Scholar
  12. Choung YH, Taura A, Pak K, Choi SJ, Masuda M, Ryan AF (2009) Generation of highly-reactive oxygen species is closely related to hair cell damage in rat organ of Corti treated with gentamicin. Neuroscience 161:214–226PubMedCrossRefGoogle Scholar
  13. Coling D, Chen S, Chi LH, Jamesdaniel S, Henderson D (2009) Age-related changes in antioxidant enzymes related to hydrogen peroxide metabolism in rat inner ear. Neurosci Lett 464:22–25PubMedCrossRefGoogle Scholar
  14. Davis GL, Hawrisiak MM (1977) Experimental cytomegalovirus infection and the developing mouse inner ear: in vivo and in vitro studies. Lab Invest 37:20–29PubMedGoogle Scholar
  15. Dinh CT, Van De Water TR (2009) Blocking pro-cell-death signal pathways to conserve hearing. Audiol Neurootol 14:383–392CrossRefGoogle Scholar
  16. Fowler KB, McCollister FP, Dahle AJ, Boppana S, Britt WJ, Pass RF (1997) Progressive and fluctuating sensorineural hearing loss in children with asymptomatic congenital cytomegalovirus infection. J Pediatr 130:624–630PubMedCrossRefGoogle Scholar
  17. Funai H, Funasaka S (1983) Experimental study on the effect of inferior colliculus lesions upon auditory brain stem response. Audiology 22:9–19PubMedCrossRefGoogle Scholar
  18. Grosse SD, Ross DS, Dollard SC (2008) Congenital cytomegalovirus (CMV) infection as a cause of permanent bilateral hearing loss: a quantitative assessment. J Clin Virol 41:57–62PubMedCrossRefGoogle Scholar
  19. Henderson D, Bielefeld EC, Harris KC, Hu BH (2006) The role of oxidative stress in noise-induced hearing loss. Ear Hear 27:1–19PubMedCrossRefGoogle Scholar
  20. Hirose K, Discolo CM, Keasler JR, Ransohoff R (2005) Mononuclear phagocytes migrate into the murine cochlea after acoustic trauma. J Comp Neurol 489:180–194PubMedCrossRefGoogle Scholar
  21. Ichimiya I, Yoshida K, Hirano T, Suzuki M, Mogi G (2000) Significance of spiral ligament fibrocytes with cochlear inflammation. Int J Pediatr Otorhinolaryngol 56:45–51PubMedCrossRefGoogle Scholar
  22. Jeong SW, Kim LS, Hur D et al (2010) Gentamicin-induced spiral ganglion cell death: apoptosis mediated by ROS and the JNK signaling pathway. Acta Otolaryngol 130(6):670–678PubMedCrossRefGoogle Scholar
  23. Katano H, Sato Y, Tsutsui Y, Sata T, Maeda A, Nozawa N, Inoue N, Nomura Y, Kurata T (2007) Pathogenesis of cytomegalovirus-associated labyrinthitis in a guinea pig model. Microbes Infect 9:183–191PubMedCrossRefGoogle Scholar
  24. Kim HJ, Lee JH, Kim SJ et al (2010) HS Roles of NADPH oxidases in cisplatin-induced reactive oxygen species generation and ototoxicity. J Neurosci 30:3933–3946PubMedCrossRefGoogle Scholar
  25. Kim SJ, Park C, Han AL, Youn MJ, Lee JH, Kim Y, Kim ES, Kim HJ, Kim JK, Lee HK, Chung SY, So H, Park R (2009) Ebselen attenuates cisplatin-induced ROS generation through Nrf2 activation in auditory cells. Hear Res 251:70–82PubMedCrossRefGoogle Scholar
  26. Kosugi I, Kawasaki H, Arai Y, Tsutsui Y (2002) Innate immune responses to cytomegalovirus infection in the developing mouse brain and their evasion by virus-infected neurons. Am J Pathol 161:919–928PubMedCrossRefGoogle Scholar
  27. Li L, Kosugi I, Han GP, Kawasaki H, Arai Y, Takeshita T, Tsutsui Y (2008) Induction of cytomegalovirus-infected labyrinthitis in newborn mice by lipopolysaccharide: a model for hearing loss in congenital CMV infection. Lab Invest 88:722–730PubMedCrossRefGoogle Scholar
  28. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  29. Low WK, Sun L, Tan MG, Chua AW, Wang DY (2008) L-N-Acetylcysteine protects against radiation-induced apoptosis in a cochlear cell line. Acta Otolaryngol 128:440–445PubMedCrossRefGoogle Scholar
  30. Marques CP, Cheeran MC, Palmquist JM, Hu S, Lokensgard JR (2008) Microglia are the major cellular source of inducible nitric oxide synthase during experimental herpes encephalitis. J Neurovirol 14:229–238PubMedCrossRefGoogle Scholar
  31. Matkovic S, Vojvodic D, Baljosevic I (2007) Comparison of cytokine levels in bilateral ear effusions in patients with otitis media secretoria. Otolaryngol Head Neck Surg 137:450–453PubMedCrossRefGoogle Scholar
  32. Micheau O, Tschopp J (2003) Induction of TNF receptor I-mediated apoptosis via two sequential signaling complexes. Cell 114:181–190PubMedCrossRefGoogle Scholar
  33. Muppidi JR, Tschopp J, Siegel RM (2004) Life and death decisions: secondary complexes and lipid rafts in TNF receptor family signal transduction. Immunity 21:461–465PubMedCrossRefGoogle Scholar
  34. Mutnal MB, Cheeran MC, Hu S, Lokensgard JR (2011) Murine cytomegalovirus infection of neural stem cells alters neurogenesis in the developing brain. PLoS One 6:e16211.Google Scholar
  35. Nesin M, Cunningham-Rundles S (2000) Cytokines and neonates. Am J Perinatol 17:393–404PubMedCrossRefGoogle Scholar
  36. Ohinata Y, Miller JM, Altschuler RA, Schacht J (2000) Intense noise induces formation of vasoactive lipid peroxidation products in the cochlea. Brain Res 878:163–173PubMedCrossRefGoogle Scholar
  37. Ohinata Y, Miller JM, Schacht J (2003) Protection from noise-induced lipid peroxidation and hair cell loss in the cochlea. Brain Res 966:265–273PubMedCrossRefGoogle Scholar
  38. Otis EM, Brent R (1954) Equivalent ages in mouse and human embryos. Anat Rec 120:33–63PubMedCrossRefGoogle Scholar
  39. Park AH, Gifford T, Schleiss MR et al (2010) Development of cytomegalovirus-mediated sensorineural hearing loss in a Guinea pig model. Arch Otolaryngol Head Neck Surg 136:48–53PubMedCrossRefGoogle Scholar
  40. Satoh H, Firestein GS, Billings PB, Harris JP, Keithley EM (2002) Tumor necrosis factor-alpha, an initiator, and etanercept, an inhibitor of cochlear inflammation. Laryngoscope 112:1627–1634PubMedCrossRefGoogle Scholar
  41. Schachtele SJ, Hu S, Little MR et al (2010) Herpes simplex virus induces neural oxidative damage via microglial cell Toll-like receptor-2. J Neuroinflammation 7:35PubMedCrossRefGoogle Scholar
  42. Song L, McGee J, Walsh EJ (2006) Frequency- and level-dependent changes in auditory brainstem responses (ABRS) in developing mice. J Acoust Soc Am 119:2242–2257PubMedCrossRefGoogle Scholar
  43. Staczek J (1990) Animal cytomegaloviruses. Microbiol Rev 54:247–265PubMedGoogle Scholar
  44. Stoddart CA, Cardin RD, Boname JM, Manning WC, Abenes GB, Mocarski ES (1994) Peripheral blood mononuclear phagocytes mediate dissemination of murine cytomegalovirus. J Virol 68:6243–6253PubMedGoogle Scholar
  45. van Den Pol AN, Mocarski E, Saederup N, Vieira J, Meier TJ (1999) Cytomegalovirus cell tropism, replication, and gene transfer in brain. J Neurosci 19:10948–10965Google Scholar
  46. Van Wijk F, Staecker H, Keithley E, Lefebvre PP (2006) Local perfusion of the tumor necrosis factor alpha blocker infliximab to the inner ear improves autoimmune neurosensory hearing loss. Audiol Neurootol 11:357–365PubMedCrossRefGoogle Scholar
  47. Wakabayashi K, Fujioka M, Kanzaki S et al (2009) Blockade of interleukin-6 signaling suppressed cochlear inflammatory response and improved hearing impairment in noise-damaged mice cochlea. Neurosci Res 66(4):345–352PubMedCrossRefGoogle Scholar
  48. Wang X, Truong T, Billings PB, Harris JP, Keithley EM (2003) Blockage of immune-mediated inner ear damage by etanercept. Otol Neurotol 24:52–57PubMedCrossRefGoogle Scholar
  49. Whitlon DS, Szakaly R, Greiner MA (2001) Cryoembedding and sectioning of cochleas for immunocytochemistry and in situ hybridization. Brain Res Brain Res Protoc 6:159–166PubMedCrossRefGoogle Scholar
  50. Yamashita D, Jiang HY, Schacht J, Miller JM (2004) Delayed production of free radicals following noise exposure. Brain Res 1019:201–209PubMedCrossRefGoogle Scholar
  51. Zine A, van de Water TR (2004) The MAPK/JNK signalling pathway offers potential therapeutic targets for the prevention of acquired deafness. Curr Drug Targets CNS Neurol Disord 3:325–332PubMedCrossRefGoogle Scholar

Copyright information

© Journal of NeuroVirology, Inc. 2011

Authors and Affiliations

  • Scott J. Schachtele
    • 1
  • Manohar B. Mutnal
    • 1
  • Mark R. Schleiss
    • 2
  • James R. Lokensgard
    • 1
    • 3
    Email author
  1. 1.Center for Infectious Diseases and Microbiology Translational Research, Department of MedicineUniversity of MinnesotaMinneapolisUSA
  2. 2.Center for Infectious Diseases and Microbiology Translational Research, Department of PediatricsUniversity of MinnesotaMinneapolisUSA
  3. 3.3-430 Translational Research FacilityUniversity of MinnesotaMinneapolisUSA

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