1887

Abstract

Equine herpesvirus type 1 (EHV-1) is an emerging pathogen that causes encephalomyelitis in horses and non-equid species. Several aspects of the immune response in the central nervous system (CNS), mainly regarding the role of inflammatory mediators during EHV-1 encephalitis, remain unknown. Moreover, understanding the mechanisms underlying extensive neuropathology induced by viruses would be helpful to establish therapeutic strategies. Therefore, we aimed to evaluate some aspects of the innate immune response during highly neurovirulent EHV-1 infection. C57BL/6 mice infected intranasally with A4/72 and A9/92 EHV-1 strains developed a fulminant neurological disease at 3 days post-inoculation with high viral titres in the brain. These mice developed severe encephalitis with infiltration of monocytes and CD8 T cells to the brain. The inflammatory infiltrate followed the detection of the chemokines CCL2, CCL3, CCL4, CCL5, CXCL2, CXCL9 and CXCL-10 in the brain. Notably, the levels of CCL3, CCL4, CCL5 and CXCL9 were higher in A4/72-infected mice, which presented higher numbers of inflammatory cells within the CNS. Pro-inflammatory cytokines, such as interleukins (ILs) IL-1α, IL-1β, IL-6, IL-12β, and tumour necrosis factor (TNF), were also detected in the CNS, and Toll-like receptor (TLR) TLR2, TLR3 and TLR9 genes were also upregulated within the brain of EHV-1-infected mice. However, no expression of interferon-γ (IFN-γ) and IL-12α, which are important for controlling the replication of other herpesviruses, was detected in EHV-1-infected mice. The results show that the activated innate immune mechanisms could not prevent EHV-1 replication within the CNS, but most likely contributed to the extensive neuropathology. The mouse model of viral encephalitis proposed here will also be useful to study the mechanisms underlying extensive neuropathology.

Funding
This study was supported by the:
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2016/24856-0)
    • Principle Award Recipient: PauloC. Maiorka
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2016/09396-2)
    • Principle Award Recipient: LeonardoPereira Mesquita
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/content/journal/jgv/10.1099/jgv.0.001556
2021-02-02
2021-10-26
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References

  1. Ostlund E. The equine herpesviruses. Vet Clin North Am - Equine Pract 1993; 9:283–294
    [Google Scholar]
  2. Pusterla N, Hussey GS. Equine herpesvirus 1 myeloencephalopathy. Vet Clin North Am - Equine Pract 2014; 30:489–506
    [Google Scholar]
  3. Kydd JH, Smith KC, Hannant D, Livesay GJ, Mumford JA. Distribution of equid herpesvirus‐1 (EHV‐1) in respiratory tract associated lymphoid tissue: implications for cellular immunity. Equine Vet J 1994; 26:470–473
    [Google Scholar]
  4. Kydd JH, Smith KC, Hannant D, Livesay GJ, Mumford JA. Distribution of equid herpesvirus‐1 (EHV‐1) in the respiratory tract of ponies: implications for vaccination strategies. Equine Vet J 1994; 26:466–469
    [Google Scholar]
  5. Goehring LS, Soboll Hussey G, Gomez Diez M, Benedict K, Maxwell LK et al. Plasma D-dimer concentrations during experimental EHV-1 infection of horses. J Vet Intern Med 2013; 27:1535–1542
    [Google Scholar]
  6. Allen GP. Risk factors for development of neurologic disease after experimental exposure to equine herpesvirus-1 in horses. Am J Vet Res 2008; 69:1595–1600
    [Google Scholar]
  7. Edington N, Bridges C, Patel J. Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: equine stroke. Arch Virol 1986; 90:111–124
    [Google Scholar]
  8. Wohlsein P, Lehmbecker A, Spitzbarth I, Algermissen D, Baumgärtner W et al. Fatal epizootic equine herpesvirus 1 infections in new and unnatural hosts. Vet Microbiol 2011; 149:456–460
    [Google Scholar]
  9. Greenwood AD, Tsangaras K, SYW H, Szentiks CA, Nikolin VM et al. A potentially fatal mix of herpes in zoos. Curr Biol 2012; 22:1727–1731
    [Google Scholar]
  10. Awan AR, Chong YC, Field HJ. The pathogenesis of equine herpesvirus type 1 in the mouse: a new model for studying host responses to the infection. J Gen Virol 1990; 71:1131–1140
    [Google Scholar]
  11. Baxi MK, Borchers K, Bartels T, Schellenbach A, Baxi S et al. Molecular studies of the acute infection, latency and reactivation of equine herpesvirus-1 (EHV-1) in the mouse model. Virus Res 1996; 40:33–45
    [Google Scholar]
  12. Kim SK, Shakya AK. Immunization with attenuated equine herpesvirus 1 strain KyA; 2016; 908090–8104
  13. Van de Walle GR, Sakamoto K, Osterrieder N. CCL3 and viral chemokine-binding protein GG modulate pulmonary inflammation and virus replication during equine herpesvirus 1 infection. J Virol 2008; 82:1714–1722 [View Article][PubMed]
    [Google Scholar]
  14. Zanuzzi C, Scrochi M, Fuentealba N, Nishida F, Portiansky E et al. Effects of equid herpesvirus 1 (EHV-1) AR8 and HH1 strains on BALB-c mice. Arch Virol 2014; 159:141–145
    [Google Scholar]
  15. Hasebe R, Kimura T, Sato E, Okazaki K, Ochiai K et al. Equine herpesvirus-1-induced encephalomyelitis in mice: a comparative study of neuroadapted virus and its parental strain. J Comp Pathol 2002; 127:118–125
    [Google Scholar]
  16. Plummer G, Coleman PL, Cleveland PH. Neurovirulence of equine herpesvirus type 1. J Infect Dis 1973; 128:202–210
    [Google Scholar]
  17. Patel J, Edington N. The pathogenicity in mice of respiratory, abortion and paresis isolates of equine herpesvirus-1. Vet Microbiol 1983; 8:301–305
    [Google Scholar]
  18. Frampton AR, Smith PM, Zhang Y, Grafton WD, Matsumura T et al. Meningoencephalitis in mice infected with an equine herpesvirus 1 strain KyA recombinant expressing glycoprotein I and glycoprotein E. Virus Genes 2004; 29:9–17
    [Google Scholar]
  19. Gosztonyi G, Borchers K, Ludwig H. Pathogenesis of equine herpesvirus-1 infection in the mouse model. Apmis 2009; 117:10–21
    [Google Scholar]
  20. Mesquita LP, Arévalo AF, Zanatto DA, Miyashiro SI, Cunha EMS et al. Equine herpesvirus type 1 induces both neurological and respiratory disease in Syrian hamsters. Vet Microbiol 2017; 203:
    [Google Scholar]
  21. Mori CMC, Mori E, Favaro LL, Santos CR, Lara M et al. Equid herpesvirus type-1 exhibits neurotropism and neurovirulence in a mouse model. J Comp Pathol 2012; 146:202–210
    [Google Scholar]
  22. Nair S, Diamond MS. Innate immune interactions within the central nervous system modulate pathogenesis of viral infections. Curr Opin Immunol 2015; 36:47–53
    [Google Scholar]
  23. Becher B, Spath S, Goverman J. Cytokine networks in neuroinflammation. Nat Rev Immunol 2017; 17:49–59
    [Google Scholar]
  24. Conrady CD, Drevets DA, Carr DJJ. Herpes simplex type I (HSV-1) infection of the nervous system: is an immune response a good thing?. J Neuroimmunol 2010; 220:1–9
    [Google Scholar]
  25. Lokensgard JR, Hu S, Sheng W, vanOijen M, Cox D et al. Robust expression of TNF-α, IL-1β, RANTES, and IP-10 by human microglial cells during nonproductive infection with herpes simplex virus. J Neurovirol 2001; 7:208–219
    [Google Scholar]
  26. Shives KD, Tyler KL, Beckham JD. Molecular mechanisms of neuroinflammation and injury during acute viral encephalitis. J Neuroimmunol 2017; 308:102–111
    [Google Scholar]
  27. Pusterla N, Wilson WD, Conrad PA, Barr BC, Ferraro GL et al. Cytokine gene signatures in neural tissue of horses with equine protozoal myeloencephalitis or equine herpes type 1 myeloencephalopathy. Vet Rec 2006; 159:341–346
    [Google Scholar]
  28. Holz CL, Nelli RK, Eilidh Wilson M, Zarski LM, Azab W et al. Viral genes and cellular markers associated with neurological complications during herpesvirus infections. J Gen Virol 2017; 98:1439–1454
    [Google Scholar]
  29. Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity. Immunity 2000; 1:121–127
    [Google Scholar]
  30. Vilela MC, Mansur DS, Lacerda-Queiroz N, Rodrigues DH, Lima GK et al. The chemokine CCL5 is essential for leukocyte recruitment in a model of severe herpes simplex encephalitis. Ann N Y Acad Sci 2009; 1153:256–263
    [Google Scholar]
  31. Aravalli RN, Hu S, Rowen TN, Palmquist JM, Lokensgard JR. Cutting edge: TLR2-mediated proinflammatory cytokine and chemokine production by microglial cells in response to herpes simplex virus. J Immunol 2005; 175:4189–4193
    [Google Scholar]
  32. Nugent J, Smith KC, Mumford JA, Newton JR, Bowden RJ et al. Analysis of equid herpesvirus 1 strain variation reveals a point mutation of the DNA polymerase strongly associated with neuropathogenic versus nonneuropathogenic disease outbreaks. J Virol 2006; 80:4047–4060 [View Article][PubMed]
    [Google Scholar]
  33. Reed L, Muench H. A simple method of estimating fifty percent endopoints. Am J Epidemiol 1938; 27:493–497
    [Google Scholar]
  34. Bolanle F, Yongshan M, Maria S, Modinat L, Hallenbeck J. Downstream Toll-like receptor signaling mediates adaptor-specific cytokine expression following focal cerebral ischemia. J Neuroinflammation 2012; 9:1–11
    [Google Scholar]
  35. Cardona AE, Huang DR, Sasse ME, Ransohoff RM. Isolation of murine microglial cells for RNA analysis or flow cytometry. Nat Protoc 2006; 1:1947–1951
    [Google Scholar]
  36. Burguillos MA. Microglia isolation. Microglia Methods Protoc Methods Mol Biol 2013; 1041:93–100
    [Google Scholar]
  37. Michlmayr D, McKimmie CS, Pingen M, Haxton B, Mansfield K et al. Defining the chemokine basis for leukocyte recruitment during viral encephalitis. J Virol 2014; 88:9553–9567
    [Google Scholar]
  38. Steibel JP, Poletto R, Coussens PM, Rosa GJM. A powerful and flexible linear mixed model framework for the analysis of relative quantification RT-PCR data. Genomics 2009; 94:146–152
    [Google Scholar]
  39. Sedgwick JD, Schwender S, Imrich H, Dorries R, Butcher GW et al. Isolation and direct characterization of resident microglial cells from the normal and inflamed central nervous system. Proc Natl Acad Sci U S A 1991; 88:7438–7442
    [Google Scholar]
  40. Klein RS, Hunter CA. Protective and pathological immunity during central nervous system infections. Immunity 2017; 46:891–909
    [Google Scholar]
  41. Terry RL, Getts DR, Deffrasnes C, van Vreden C, Campbell IL et al. Inflammatory monocytes and the pathogenesis of viral encephalitis. J Neuroinflammation 2012; 9:1–10
    [Google Scholar]
  42. Shrestha B, Diamond MS. Role of CD8+ T cells in control of West Nile virus infection. J Virol 2004; 78:8312–8321
    [Google Scholar]
  43. Lang A, Nikolich-Žugich J. Development and migration of protective CD8 + T cells into the nervous system following ocular herpes simplex virus-1 infection. J Immunol 2005; 174:2919–2925
    [Google Scholar]
  44. Griffin DE. Recovery from viral encephalomyelitis: immune-mediated noncytolytic virus clearance from neurons. Immunol Res 2010; 47:123–133
    [Google Scholar]
  45. Koyanagi N, Imai T, Shindo K, Sato A, Fujii W et al. Herpes simplex virus-1 evasion of CD8+ T cell accumulation contributes to viral encephalitis. J Clin Invest 2017; 127:3784–3795
    [Google Scholar]
  46. Allen G, Yeargan M, Costa LR, Cross R. Major histocompatibility complex class I-restricted cytotoxic T-lymphocyte responses in horses infected with equine herpesvirus 1. J Virol 1995; 69:606–612
    [Google Scholar]
  47. Smith PM, Zhang Y, Jennings SR, O’Callaghan DJ. Characterization of the cytolytic T-lymphocyte response to a candidate vaccine strain of equine herpesvirus 1 in CBA mice. J Virol 1998; 72:5366–5372
    [Google Scholar]
  48. Liu SA, Stanfield BA, Chouljenko VN, Naidu S, Langohr I et al. Intramuscular immunization of mice with the live-attenuated herpes simplex virus 1 vaccine strain VC2 expressing equine herpesvirus 1 (EHV-1) glycoprotein D generates Anti-EHV-1 immune responses in mice. J Virol 2017; 91:1–15
    [Google Scholar]
  49. Marques CP, Hu S, Sheng W, Lokensgard JR. Microglial cells initiate vigorous yet non-protective immune responses during HSV-1 brain infection. Virus Res 2006; 121:1–10
    [Google Scholar]
  50. Kumar M, Verma S, Nerurkar VR. Pro-inflammatory cytokines derived from West Nile virus (WNV)-infected SK-N-SH cells mediate neuroinflammatory markers and neuronal death. J Neuroinflammation 2010; 7:1–14
    [Google Scholar]
  51. Brabers N, Nottet H. Role of the pro-inflammatory cytokines TNF- α and IL-1 β in HIV-associated dementia; 2006447–458
  52. Chen CJ, YC O, Lin SY, Raung SL, Liao SL et al. Glial activation involvement in neuronal death by Japanese encephalitis virus infection. J Gen Virol 2010; 91:1028–1037
    [Google Scholar]
  53. Das S, Mishra MK, Ghosh J, Basu A. Japanese encephalitis virus infection induces IL-18 and IL-1β in microglia and astrocytes: correlation with in vitro cytokine responsiveness of glial cells and subsequent neuronal death. J Neuroimmunol 2008; 195:60–72
    [Google Scholar]
  54. Campbell IL, Abraham CR, Masliah E, Kemper P, Inglis JD et al. Neurologic disease induced in transgenic mice by cerebral overexpression of interleukin 6. Proc Natl Acad Sci U S A 1993; 90:10061–10065
    [Google Scholar]
  55. Wang T, Town T, Alexopoulou L, Anderson JF, Fikrig E et al. Toll-Like receptor 3 mediates West Nile virus entry into the brain causing lethal encephalitis. Nat Med 2004; 10:1366–1373
    [Google Scholar]
  56. Ramakrishna C, Cantin EM. IFNγ inhibits G-CSF induced neutrophil expansion and invasion of the CNS to prevent viral encephalitis. PLoS Pathog 2018; 14:1–27
    [Google Scholar]
  57. Lekstrom-Himes JA, LeBlanc RA, Pesnicak L, Godleski M, Straus SE. Gamma interferon impedes the establishment of herpes simplex virus type 1 latent infection but has no impact on its maintenance or reactivation in mice. J Virol 2000; 74:6680–6683
    [Google Scholar]
  58. Lin AA, Tripathi PK, Sholl A, Jordan MB, Hildeman DA. Gamma interferon signaling in macrophage lineage cells regulates central nervous system inflammation and chemokine production. J Virol 2009; 83:8604–8615
    [Google Scholar]
  59. McCarthy MK, Procario MC, Twisselmann N, Wilkinson JE, Archambeau AJ et al. Proinflammatory effects of interferon gamma in mouse adenovirus 1 myocarditis. J Virol 2015; 89:468–479
    [Google Scholar]
  60. O’Flaherty BM, Matar CG, Wakeman BS, Garcia AP, Wilke CA et al. Cd8+ T cell response to gammaherpesvirus infection mediates inflammation and fibrosis in interferon gamma receptor-deficient mice. PLoS One 2015; 10:1–25
    [Google Scholar]
  61. Schroder K, Hertzog P. Ifn gamma an overview. J Leukoc Biol 2004; 75:163–189
    [Google Scholar]
  62. Stubblefield Park SR, Widness M, Levine AD, Patterson CE, Cell- T. Interleukin-12-, and gamma interferon-driven viral clearance in measles virus-infected brain tissue. J Virol 2011; 85:3664–3676
    [Google Scholar]
  63. Soboll Hussey G, Ashton LV, Quintana AM, Lunn DP, Goehring LS et al. Innate immune responses of airway epithelial cells to infection with equine herpesvirus-1. Vet Microbiol 2014; 170:28–38 [View Article][PubMed]
    [Google Scholar]
  64. Hosking MP, Lane TE. The role of chemokines during viral infection of the CNS. PLoS Pathog 2010; 6:1–4
    [Google Scholar]
  65. Smith PM, Kahan SM, Rorex CB, von Einem J, Osterrieder N et al. Expression of the full-length form of GP2 of equine herpesvirus 1 (EHV-1) completely restores respiratory virulence to the attenuated EHV-1 strain KyA in CBA mice. J Virol 2005; 79:5105–5115 [View Article][PubMed]
    [Google Scholar]
  66. Smith PM, Zhang Y, Grafton WD, Jennings SR, O'Callaghan DJ. Severe murine lung immunopathology elicited by the pathogenic equine herpesvirus 1 strain RacL11 correlates with early production of macrophage inflammatory proteins 1alpha, 1beta, and 2 and tumor necrosis factor alpha. J Virol 2000; 74:10034–10040 [View Article][PubMed]
    [Google Scholar]
  67. Soboll Hussey G, Ashton LV, Quintana AM, Van de Walle GR, Osterrieder N et al. Equine herpesvirus type 1 pUL56 modulates innate responses of airway epithelial cells. Virology 2014; 464-465:76–86 [View Article][PubMed]
    [Google Scholar]
  68. Zhao J, Poelaert KCK, Van Cleemput J, Nauwynck HJ. CCL2 and CCL5 driven attraction of CD172a+ monocytic cells during an equine herpesvirus type 1 (EHV-1) infection in equine nasal mucosa and the impact of two migration inhibitors, rosiglitazone (RSG) and quinacrine (Qc). Vet Res 2017; 48:1–12
    [Google Scholar]
  69. Poelaert KCK, Van CJ, Laval K, Xie J, Favoreel HW. Equine herpesvirus 1 infection orchestrates the expression of chemokines in equine respiratory epithelial cells; 20191567–1579
  70. Johnstone S, Barsova J, Campos I, Frampton AR. Equine herpesvirus type 1 modulates inflammatory host immune response genes in equine endothelial cells. Vet Microbiol 2016; 192:52–59
    [Google Scholar]
  71. Wimer CL, Damiani A, Osterrieder N, Wagner B. Equine herpesvirus type-1 modulates CCL2, CCL3, CCL5, CXCL9, and CXCL10 chemokine expression. Vet Immunol Immunopathol 2011; 140:266–274
    [Google Scholar]
  72. Paludan SR, Bowie AG, Horan KA, Fitzgerald KA. Recognition of herpesviruses by the innate immune system. Nat Rev Immunol 2011; 11:143–154
    [Google Scholar]
  73. Wang JP, Bowen GN, Zhou S, Cerny A, Zacharia A et al. Role of specific innate immune responses in herpes simplex virus infection of the central nervous system. J Virol 2012; 86:2273–2281
    [Google Scholar]
  74. Guo Y, Audry M, Ciancanelli M, Alsina L, Azevedo J et al. Herpes simplex virus encephalitis in a patient with complete TLR3 deficiency: TLR3 is otherwise redundant in protective immunity. J Exp Med 2011; 208:2083–2098
    [Google Scholar]
  75. Zhang SY, Jouanguy E, Ugolini S, Smahi A, Elain G et al. Tlr3 deficiency in patients with herpes simplex encephalitis. Science 2007; 317:1522–1527
    [Google Scholar]
  76. Davey GM, Wojtasiak M, Proietto AI, Carbone FR, Heath WR et al. Cutting edge: priming of CD8 T cell immunity to herpes simplex virus type 1 requires cognate TLR3 expression in vivo. J Immunol 2010; 184:2243–2246 [View Article][PubMed]
    [Google Scholar]
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