Skip to main content
PMC home page
Wiley - PMC COVID-19 Collection logoLink to Wiley - PMC COVID-19 Collection
. 2013 Nov 14;22(7):377–390. doi: 10.1111/j.1348-0421.1978.tb00384.x

Physico‐Chemical Properties of Mouse Hepatitis Virus (MHV‐2) Grown on DBT Cell Culture

Norio Hirano 1,2,✉, Shigeo Hino 1,2, Kosaku Fujiwara 1,2
PMCID: PMC7168437  PMID: 30881

Abstract

Some properties of a strain of mouse hepatitis virus, MHV‐2, grown on DBT cells were determined using a plaque assay on the cells. Viral growth was not inhibited by the presence of actinomycin D or 5‐iodo‐2‐deoxyuridine. MHV‐2 was completely inactivated by ether, chloroform, sodium deoxycholate or beta‐propiolactone, but showed a moderate resistance to trypsin. Heating at 56 C for 30 min did not completely abolish the virus infectivity. The virus was stable after heating at 50 C for 15 min in 1m‐MgCl2 or 1m‐MgSO4 as well as at 37 C for 60 min at pH 3.0 to 9.0. Infectivity was decreased to 1/100 and 1/400 after storing at 4 C for 30 days and 37 C for 24 hr, respectively. The virus passed through a 200‐nm but not a 50‐nm Sartorius membrane filter. The buoyant density of MHV‐2 was 1.183 g/cm3 in sucrose gradient, and the fraction contained coronavirus‐like particles measuring 70 to 130 nm in diameter. Survival rate was 10% after exposure to ultraviolet at 150 ergs/mm2. Freezing and thawing or sonication at 20 kc for 3 min did not affect the virus titer. No hemagglutinin was demonstrable with red blood cells of the chicken, Japanese quail, mouse, rat, hamster, guinea pig, sheep, bovine or human.

REFERENCES

  • 1. Bhatt, P.N. , Percy, D.H. , and Jonas, A.M. 1972. Characterization of the virus of sialodacryoadenitis of rats. J. Infect. Dis. 126: 123–230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Bingham, R.W. , Hilary, M.M. , and Tyrrell, D.A.J. 1975. Haemagglutination by avian infectious bronchitis virus, a coronavirus. J. Gen. Virol. 28: 381–390. [DOI] [PubMed] [Google Scholar]
  • 3. Binn, L.N. , Lazar, E.C. , Keenan, K.P. , Huxsoll, D.L. , Marchwichi, R.H. , and Strano, A.J. 1974. Recovery and characterization of a coronavirus from military dogs with diarrhea. p. 359–366. In Proceeding, 78th Anm Meeting, US Anim. Health Assoc., Roanoke, Va. (Oct. 1973). [PubMed] [Google Scholar]
  • 4. Bradburne, A.F. , and Tyrrell, D.A.J. 1969. Coronaviruses of man. Prog. Med. Virol. 13: 373–403. [Google Scholar]
  • 5. Calisher, C.H. , and Rowe, W.P. 1966. In symposium on viruses of laboratory rodents. Natl. Cancer Inst. Monogr. 20: 67–75. [PubMed] [Google Scholar]
  • 6. Cartwright, F.S. , Harris, H.M. , and Blandfors, T.B. 1965. A cytopathic virus causing a transmissible gastroenteritis in swine. J. Comp. Pathol. 75: 387–396. [DOI] [PubMed] [Google Scholar]
  • 7. Cheever, F.S. , Daniels, J.B. , Pappenheimer, A.M. , and Bailey, O.T. 1949. A murine virus (JHM) causing disseminated encephalomyelitis with extensive destruction of myelin. I. Isolation and biologic properties of the virus. J. Exp. Med. 90: 181–194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. Cowen, B.S. , and Hitchner, S.H. 1975. pH studies with avian infectious bronchitis virus (coronavirus) strains. J. Virol. 15: 430–432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Coria, M.F. 1972. Stabilizing effect of magnesium sulfate on avian infectious bronchitis virus propagated in chicken embryo kidney cells. Appl. Microbiol. 23: 281–284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10. Cunningham, C.H. 1970. Avian infectious bronchitis. Adv. Vet. Comp. Med. 14: 105–148. [PubMed] [Google Scholar]
  • 11. Dick, G.W.A. , Niven, J.S.F. , and Gledhill, A.W. 1956. A virus related to that causing hepatitis in mice (MHV). Brit. J. Exp. Pathol. 37: 90–98. [PMC free article] [PubMed] [Google Scholar]
  • 12. Dulbecco, R. , and Vogt, M. 1954. Plaque formation and isolation of pure lines of poliomyelitis viruses. J. Exp. Med. 99: 167–182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. Furuuchi, S. , Shimizu, Y. , and Kumagai, T. 1975. Comparison of properties between virulent and attenuated strains of transmissible gastroenteritis virus. Natl. Inst. Anim. Health Q. 15: 159–164. [PubMed] [Google Scholar]
  • 14. Gledhill, A.W. , and Andrewes, C.H. 1951. A hepatitis virus of mice. Brit. J. Exp. Pathol. 32: 559–568. [PMC free article] [PubMed] [Google Scholar]
  • 15. Gledhill, A.W. , Dick, G.W.A. , and Niven, J.S.F. 1955. Mouse hepatitis virus its pathogenic action. J. Pathol. Bacteriol. 66: 299–309. [DOI] [PubMed] [Google Scholar]
  • 16. Hamparian, V.V. , Hileman, M.R. , and Ketler, A. 1963. Contributions to characterization and classification of animal viruses. Proc. Soc. Exp. Biol. Med. 112: 1040–1050. [DOI] [PubMed] [Google Scholar]
  • 17. Harada, K. , Bohl, E.H. , and Kumagai, T. 1967. A mutant of the transmissible gastroenteritis virus in pigs with thermoresistance in MgCl2 . Virus 17: 49–51 (in Japanese). [Google Scholar]
  • 18. Harada, K. , Kumagai, T. , and Sasahara, J. 1963. Cytopathogenicity of transmissble gastroenteritis virus in pigs. Natl. Inst. Anim. Health Q. 3: 166–167. [PubMed] [Google Scholar]
  • 19. Hartley, J.W. , and Rowe, W.P. 1963. Tissue culture cytopathic and plaque assays for mouse hepatitis viruses. Proc. Soc. Exp. Biol. Med. 113: 403–406. [DOI] [PubMed] [Google Scholar]
  • 20. Hercules, J.H. , and Von Kaemel, E.A. 1971. The inactivation of murine viral antigens with beta‐propiolactone. Lab. Anim. Sci. 21: 497–501. [PubMed] [Google Scholar]
  • 21. Hirano, N. , Fujiwara, K. , Hino, S. , and Matumoto, M. 1974. Replication and plaque formation of mouse hepatitis virus (MHV‐2) in mouse cell line DBT culture. Arch. Ges. Virusforsch. 44: 298–302. [DOI] [PubMed] [Google Scholar]
  • 22. Hirano, N. , Fujiwara, K. , and Matumoto, M. 1976. Mouse hepatitis virus (MHV‐2). Plaque assay and propagation in mouse cell line DBT cells. Japan. J. Microbiol. 20: 219–225. [PubMed] [Google Scholar]
  • 23. Kaye, H.S. , and Dowdle, W.R. 1969. Some characteristics of hemagglutination of certain strains of IBV‐like virus. J. Infect. Dis. 120: 576–581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. Kaye, H.S. , Hierholzer, J.C. , and Dowdle, W.R. 1970. Purification and further characterization of an “TBV‐like” virus (coronavirus). Proc. Soc. Exp. Biol. (N.Y.) 135: 457–463. [DOI] [PubMed] [Google Scholar]
  • 25. Levine, S. , and Sugik, B.P. 1956. The interactions of Newcastle disease virus (NDV) with chick embryo tissue culture cells. Attachment and growth. Virology 2: 57–68. [DOI] [PubMed] [Google Scholar]
  • 26. Mallucci, L. 1965. Observations on the growth of mouse hepatitis virus (MHV‐3) in mouse macrophages. Virology 25: 30–37. [DOI] [PubMed] [Google Scholar]
  • 27. Manaker, R.A. , Piczac, C.V. , Miller, A.A. , and Stanton, M.F. 1961. A hepatitis virus complicating studies with mouse leukemia. J. Natl. Cancer Inst. 27: 27–51. [PubMed] [Google Scholar]
  • 28. McClurkin, A.W. , and Norman, J.O. 1966. Studies on transmissible gastroenteritis of swine. II. Selected characteristics of a cytopathogenic virus common to five isolates from transmissible gastroenteritis. Canad. J. Comp. Med. 30: 190–198. [PMC free article] [PubMed] [Google Scholar]
  • 29. McIntosh, K. 1974. Coronaviruses; a comparative review. Curr. Top. Microbiol. Immunol. 63: 85–129. [Google Scholar]
  • 30. Mebus, C.A. , Stair, E.L. , Rhodes, M.B. , and Twiehaus, M.J. 1973. Neonatal calf diarrhea: propagation, attenuation and characteristics of coronavirus‐like agent. Am. J. Vet. Res. 34: 145–150. [PubMed] [Google Scholar]
  • 31. Mengeling, W.L. , Boothe, A.D. , and Ritchie, A.E. 1972. Characteristics of coronavirus (strain 67) of pigs. Am. J. Vet. Res. 33: 297–308. [PubMed] [Google Scholar]
  • 32. Nelson, J.B. 1952. Acute hepatitis associated with mouse leukemia. I. Pathological features and transmission of the disease. J. Exp. Med. 96: 293–303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. Parker, J.C. , Cross, S.S. , and Rowe, W.P. 1970. Rat coronavirus (RCV): A prevalent, naturally occurring pneumotropic virus of rats. Arch. Ges. Virusforsch. 31: 293–302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. Pensaert, M.B. , and Callebaut, P.E. 1974. Characteristics of a coronavirus causing vomition and wasting in pigs. Arch. Ges. Virusforsch. 44: 35–40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. Phillip, J.I.H. , Cartwright, S.F. , and Scott, A.C. 1971. The size and morphology of T.G.E. and vomiting and wasting disease viruses of pigs. Vet. Rec. 88: 311–312. [DOI] [PubMed] [Google Scholar]
  • 36. Piazza, M. 1969. Experimental viral hepatitis. Charles C. Thomas, Illinois, U.S.A.. [Google Scholar]
  • 37. Ritchie, A.E. , Desmuke, D.R. , Larsen, C.T. , and Pomeroy, B.S. 1973. Electron microscopy of coronavirus‐like particles characteristics of turkey bluecomb disease. Avian Dis. 17: 540–558. [PubMed] [Google Scholar]
  • 38. Rowe, W.P. , Hartley, J.W. , and Capps, W.I. 1963. Mouse hepatitis virus infection as a highly contagious, prevalent, enteric infection of mice. Proc. Soc. Exp. Biol. Med. 112: 161–165. [DOI] [PubMed] [Google Scholar]
  • 39. Sever, J.L. 1962. Application of a microtechnique to viral serological investigation. J. Immunol. 88: 320–329. [PubMed] [Google Scholar]
  • 40. Stinski, M.F. , and Cunningham, C.H. 1969. Neutralizing antibody complex of infectious bronchitis virus. J. Immunol. 102: 720–727. [PubMed] [Google Scholar]
  • 41. Studdert, M.J. 1965. Sensitivity of bluetongue virus to ether and sodium deoxycholate. Proc. Soc. Exp. Biol. Med. 118: 1006–1009. [DOI] [PubMed] [Google Scholar]
  • 42. Svoboda, D. , Nielson, A. , Welder, A. , and Higginson, J. 1962. An electron microscopic study of viral hepatitis in mice. Am. J. Pathol. 41: 205–224. [PMC free article] [PubMed] [Google Scholar]
  • 43. Tajima, M. 1970. Morphology of transmissible gastroenteritis virus of pigs. Arch. Ges. Virusforsch. 29: 105–108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44. Taniguchi, S. , and Yoshino, K. 1964. An analysis of the plaque assay, for herpes simplex virus in chick embryo monolayers. Arch. Ges. Virusforsch. 14: 537–552. [DOI] [PubMed] [Google Scholar]
  • 45. Tyrrell, D.A.J. , Almeida, J.D. , Berry, D.M. , Cunningham, C.H. , Hamre, K. , Hofstad, M.S. , Mallucci, L. , and McIntosh, K. 1968. Coronaviruses. Nature (London) 220: 650. [Google Scholar]
  • 46. Tyrrell, D.A.J. , Almeida, J.D. , Cunningham, C.H. , Dowdly, M.S. , Hofstad, M.S. , McIntosh, K. , Tajima, M. , Zakstelskaya, L.Y. , Easterday, B.C. , Kapikian, A. , and Bingham, R.W. 1975. Coronaviridae. Intervirology 5: 76–82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47. Ver, A.B. , Melnick, J.L. , and Rapp, F. 1965. Efficient filtration and sizing of viruses with membrane fiters. J. Virol. 2: 21–25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48. Wallis, C. , Melnick, J.C. , and Rapp, F. 1965. Different effects of MgCl2 and MgSO4 on the thermostability of viruses. Virology 26: 694–699. [DOI] [PubMed] [Google Scholar]
  • 49. Ward, J. 1970. Morphogenesis of a virus in cats with experimental feline infectious peritonitis. Virology 41: 191–194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50. Witte, K.H. , Tajima, M. , and Easterday, B.C. 1968. Morphologic characteristics and nucleic acid type of transmissible gastroenteritis virus of pigs. Arch. Ges. Virusforsch. 23: 53–70. [DOI] [PubMed] [Google Scholar]
  • 51. Yoshikura, H. 1971. Ultraviolet inactivation of murine leukemia and sarcoma viruses. Int. J. Cancer 7: 131–140. [DOI] [PubMed] [Google Scholar]

Articles from Microbiology and Immunology are provided here courtesy of Wiley

RESOURCES