Journal of NeuroVirology

, Volume 7, Issue 5, pp 432–436 | Cite as

Sequence analysis of the S gene of recombinant MHV-2/A59 coronaviruses reveals three candidate mutations associated with demyelination and hepatitis

  • Jayasri Das Sarma
  • Li Fu
  • Susan T. Hingley
  • Michael M. C. Lai
  • Ehud Lavi


Coronaviruses, mouse hepatitis virus (MHV) strains, exhibit various degrees of neurotropism and hepatotropism following intracerebral (IC) infection of 4-week-old C57Bl/6 mice. Whereas MHV-A59 produces acute meningitis, encephalitis, hepatitis, and chronic demyelination, a closely related strain, MHV-2, produces only acute meningitis and hepatitis. We previously reported that the spike glycoprotein gene of MHV contains determinants of demyelination and hepatitis. To further investigate the site of demyelination and hepatitis determinants within the S gene, we sequenced the S gene of several nonde-myelinating recombinant viruses. We found that three encephalitis-positive, demyelination-negative, hepatitis-negative recombinant viruses have an MHV-A59-derived S gene, which contains three identical point mutations (I375M, L652I, and T1087N). One or more of the sites of these mutations in the MHV-A59 genome are likely to contribute to demyelination and hepatitis.


coronavirus nidoviruses mouse hepatitis virus (MHN) demyelination hepatitis multiple sclerosis (MS) 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Buchmeier MJ, Lewicki HA, Talbot PJ, Knobler RL (1984). Murine hepatitis virus-4 (strain JHM)-induced neurologic disease is modulated in vivo by monoclonal antibody. Virology 132: 261–270.CrossRefPubMedGoogle Scholar
  2. Cavanagh D (1997). Nidovirales: A new order comprising Coronaviridae and Arteriviridae. Arch Virol 142: 629–633.PubMedGoogle Scholar
  3. Collins AR, Knobler RL, Powell H, Buchmeier MJ (1982). Monoclonal antibodies to murine hepatitis virus-4 (strain JHM) define the viral glycoprotein responsible for attachment and cell-cell fusion. Virology 119: 358–371.CrossRefPubMedGoogle Scholar
  4. Dalziel RG, Lampert PW, Talbot PJ, Buchmeier MJ (1986). Site specific alteration of murine hepatitis virus type 4 peplomer glycoprotein S results in reduced neurovirulence. J Virol 59: 463–471.PubMedGoogle Scholar
  5. Das Sarma J, Fu L, Hingley ST, Lavi E (2001). Mouse hepatitis virus type-2 infection in mice: An experimental model system of acute meningitis and hepatitis. Exp Mol Pathol 71: 1–12.CrossRefPubMedGoogle Scholar
  6. Das Sarma J, Fu L, Tsai JC, Weiss SR, Lavi E (2000). Demyelination determinants map to the spike glycoprotein gene of coronavirus mouse hepatitis virus. J Virol 74: 9206–9213.CrossRefPubMedGoogle Scholar
  7. Fleming JO, Shubin RA, Sussman MA, Casteel N, Stohlman SA (1989). Monoclonal antibodies to the matrix (E1) glycoprotein of mouse hepatitis virus protect mice from encephalitis. Virology 168: 162–167.CrossRefPubMedGoogle Scholar
  8. Gallagher TM, Escarmis C, Buchmeier MJ (1991). Alteration of the pH dependence of coronavirus-induced cell fusion: Effect of mutations in the spike glycoprotein. J Virol 65: 1916–1928.PubMedGoogle Scholar
  9. Hingley ST, Gombold JL, Lavi E, Weiss SR (1994). MHV-A59 fusion mutants are attenuated and display altered hepatotropism. Virology 200: 1–10.CrossRefPubMedGoogle Scholar
  10. Hirano N, Fujiwara K, Hino S, Matumoto M (1974). Replication and plaque formation of mouse hepatitis virus (MHV-2) in mouse cell line DBT culture. Arch Ges Virusfors 44: 298–302.CrossRefGoogle Scholar
  11. Hirano N, Murakami T, Taguchi F, Fujiwara K, Matumoto M (1981). Comparison of mouse hepatitis virus strains for pathogenicity in weanling mice infected by various routes. Arch Virol 70: 69–73.CrossRefPubMedGoogle Scholar
  12. Houtman JJ, Fleming JO (1996). Pathogenesis of mouse hepatitis virus-induced demyelination. J Neuro Virol 2: 361–376.Google Scholar
  13. Keck JG, Soe LH, Makino S, Stohlman SA, Lai MMC (1988). RNA recombination of murine coronavirus: Recombination between fusion-positive mouse hepatitis virus A59 and fusion-negative mouse hepatitis virus 2. J Virol 62: 1989–1998.PubMedGoogle Scholar
  14. Knobler RL, Haspel MV, Oldstone MBA (1981). Mouse hepatitis virus type 4 (JHM strain)-induced fatal central nervous system disease, part 1 (genetic control and the murine neurone as the susceptible site for disease). J Exp Med 153: 832–843.CrossRefPubMedGoogle Scholar
  15. Knobler RL, Tunison LA, Lampert PW, Oldstone MBA (1982). Selected mutants of mouse hepatitis virus type 4 (JHM strain) induce different CNS diseases. Pathobiology of disease induced by wild type and mutants ts8 and ts15 in BALB/c and SJL/J mice. Am J Pathol 109: 157–168.PubMedGoogle Scholar
  16. Lai MMC, Cavanagh D (1997). The molecular biology of coronaviruses. Adv Virus Res 48: 1–100.CrossRefPubMedGoogle Scholar
  17. Laude H, Gelfi J, Lavenant L, Charley B (1992). Single amino acid changes in the viral glycoprotein M affect induction of interferon by the coronavirus transmissible gastroenteritis virus. J Virol 66: 743–749.PubMedGoogle Scholar
  18. Lavi E, Fishman SP, Highkin MK, Weiss SR (1988). Limbic encephalitis following inhalation of murine coronavirus MHV-A59. Lab Invest 58: 31–36.PubMedGoogle Scholar
  19. Lavi E, Gilden DH, Highkin MK, Weiss SR (1986). The organ tropism of mouse hepatitis virus A59 is dependent on dose and route of inoculation. Lab Anim Sci 36: 130–135.PubMedGoogle Scholar
  20. Lavi E, Gilden DH, Wroblewska Z, Rorke LB, Weiss SR (1984). Experimental demyelination produced by the A59 strain of mouse hepatitis virus. Neurology 34: 597–603.PubMedGoogle Scholar
  21. Lavi E, Haluskey JA, Masters PS (1998a). Targeted recombination between MHV-2 and MHV-A59 to study neurotropic determinants of MHV. Adv Exp Med Biol 440: 543–547.PubMedGoogle Scholar
  22. Lavi E, Kuo L, Haluskey JA, Masters PS (1998b). The pathogenesis of MHV nucleocapsid gene chimeric viruses. Adv Exp Med Biol 440: 537–541.PubMedGoogle Scholar
  23. Lavi E, Murray EM, Makino S, Stohlman SA, Lai MM, Weiss SR (1990). Determinants of coronavirus MHV pathogenesis are localized to 3′ portions of the genome as determined by ribonucleic acid-ribonucleic acid recombination. Lab Invest 62: 570–578.PubMedGoogle Scholar
  24. Lavi E, Schwartz T, Jin YP, Fu L (1999). Nidovirus infections: Experimental model systems of human neurologic diseases. J Neuropathol Exp Neurol 58: 1197–1206.CrossRefPubMedGoogle Scholar
  25. Lavi E, Weiss SR (1989). Coronaviruses. In: Clinical and molecular aspects of neurotropic viral infections. Gilden DH, Lipton HL (eds). Kluwer Academic Publishers: Boston, Massachusetts, pp 101–139.Google Scholar
  26. Leparc-Goffart I, Hingley ST, Chua M-M, Jiang X, Lavi E, Weiss SR (1997). Altered pathogenesis phenotypes of murine coronavirus MHV-A59 are associated with a Q159L amino acid substitution in the receptor binding domain of the spike protein. Virology 239: 1–10.CrossRefPubMedGoogle Scholar
  27. Leparc-Goffart I, Hingley ST, Chua MM, Phillips J, Lavi E, Weiss SR (1998). Targeted recombination within the spike gene of murine coronavirus mouse hepatitis virus-A59: Q159 is a determinant of hepatotropism. J Virol 72: 9628–9636.PubMedGoogle Scholar
  28. Navas S, Seo S, Chua MM, Das Sarma J, Lavi E, Hingley ST, Weiss SR (2001). The spike protein of murine coronavirus determines the ability of the virus to replicate in the liver and cause hepatitis. J Virol 75: 2452–2457.CrossRefPubMedGoogle Scholar
  29. Perlman S, Jacobsen G, Olson AL, Afifi A (1990). Identification of the spinal cord as a major site of persistence during chronic infection with a murine coronavirus. Virology 175: 418–426.CrossRefPubMedGoogle Scholar
  30. Phillips JJ, Chua MM, Lavi E, Weiss SR (1999). Pathogenesis of chimeric MHV-4/MHV-A59 recombinant viruses: The murine coronavirus spike protein is a major determinant of neurovirulence. J Virol 73: 7752–7760.PubMedGoogle Scholar
  31. Reed L, Muench H (1938). A simple method of estimating fifty percent end points. Am J Hyg 27: 493–497.Google Scholar
  32. Wege H, Siddell S, ter Meulen V (1982). The biology and pathogenesis of coronaviruses. Adv Virol Immunol 99: 165–200.Google Scholar
  33. Wege H, Stephenson JR, Koga M, Wege H, ter Meulen V (1981). Genetic variation of neurotropic and non-neurotropic murine coronaviruses. J Gen Virol 54: 67–74.CrossRefPubMedGoogle Scholar
  34. Weiner LP (1973). Pathogenesis of demyelination induced by a mouse hepatitis virus (JHM virus). Arch Neurol 28: 298–303.PubMedGoogle Scholar
  35. Yamada YK, Takimoto K, Yabe M, Taguchi F (1997). Acquired fusion activity of a murine coronavirus MHV-2 variant with mutations in the proteolytic cleavage site and the siganal sequence of the S protein. Virology 227: 215–219.CrossRefPubMedGoogle Scholar
  36. Yokomori K, Banner LR, Lai MC (1991). Heteroginity of gene expression of the hemagglutinin-esterase (HE) protein of murine coronaviruses. Virology 183: 647–657.CrossRefPubMedGoogle Scholar

Copyright information

© Journal of NeuroVirology, Inc. 2001

Authors and Affiliations

  • Jayasri Das Sarma
    • 1
  • Li Fu
    • 1
  • Susan T. Hingley
    • 2
  • Michael M. C. Lai
    • 3
  • Ehud Lavi
    • 1
  1. 1.Department of Pathology and Laboratory Medicine, 613 Stellar-Chance LaboratoriesUniversity of Pennsylvania, School of Medicine, Division of NeuropathologyPhiladelphiaUSA
  2. 2.Department of Microbiology and ImmunologyPhiladelphia College of Osteopathic MedicinePhiladelphia
  3. 3.Department of Molecular Microbiology and Immunology, Howard Hughes Medical InstituteUniversity of Southern California, School of MedicineLos AngelesUSA

Personalised recommendations