Archives of Virology

, Volume 137, Issue 3–4, pp 355–370 | Cite as

Loss of active neuroinvasiveness in attenuated strains of West Nile virus: pathogenicity in immunocompetent and SCID mice

  • M. Halevy
  • Y. Akov
  • D. Ben-Nathan
  • D. Kobiler
  • B. Lachmi
  • S. Lustig
Original Papers

Summary

The neuropathogenicity of West Nile virus (WNV) and two derived attenuated strains WN25 and WN25A, was studied in young adult ICR mice and in severe combined immunodeficient (SCID) mice. Similarity in serology and RNA fingerprints were found between WNV and WN25. The viral envelope proteins of the attenuates differed from WNV in their slower mobility in SDS-PAGE due probably to the presence of N-linked glycan. The three strains were lethal to ICR mice by intracerebral (IC) inoculation, but when inoculated intraperitoneally (IP), WNV caused viremia, invaded the CNS and was lethal, whereas the attenuates showed no viremia or invasion of the CNS. The attenuates elicited antibodies to comparable levels as WNV in IP-infected mice, conferring upon them immunity to IC challenge with the wild type. In IP-inoculated SCID mice the three strains exhibited similar high viremiae that lasted until death of the animals. All strains invaded the CNS and proliferated in the mouse brain to similar high titers, but differed largely in the time of invasion: WNV invaded the CNS of SCID mice (and two other mouse strains) much earlier than the attenuates, which showed large intervals in their time of invasion into individual mouse brains within the group. The data presented for SCID mice indicate that WN25 and WN25A have truly lost the neuroinvasive property, and that this property materialized by a prescribed, active process specific for WNV.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Akov Y, Halevy M, Lustig S (1987) Neuroinvasiveness of West Nile virus (WNV) strains and the use of modulators of the blood brain barrier (BBB). In: 7th International Congress of Virology, Edmonton, Alberta, Canada, p 74Google Scholar
  2. 2.
    Ben-Nathan D, Lustig S, Danenberg HD (1991) Stress-induced neuroinvasiveness of a neurovirulent noninvasive sindbis virus in cold or isolation subjected mice. Life Sci 48: 1493–1500Google Scholar
  3. 3.
    Ben-Nathan D, Danenberg HD, Halevy M, Lustig S (1992) Neuroinvasion of non-neuroinvasive attenuated arboviruses in mice subjected to stress. In: Advances in psychoneuroimmunology, 8th International Congress of Immunology, Budapest, Hungry, p 8Google Scholar
  4. 4.
    Besselaar TG, Blackburn NK (1988) Antigenic analysis of West Nile virus strains using monoclonal antibodies. Arch Virol 99: 75–88Google Scholar
  5. 5.
    Bosma GC, Custer RP, Bosma MJ (1983) A severe combined immunodeficiency mutation in the mouse. Nature 301: 527–530Google Scholar
  6. 6.
    Brinton MA (1983) Analysis of extracellular West Nile virus particles produced by cell cultures from genetically resistant and susceptible mice indicates enchanced amplification of defective interfering particles by resistant cultures. J Virol 46: 860–870Google Scholar
  7. 7.
    Cane PA, Gould EA (1989) Immunoblotting reveals differences in the accumulation of envelope protein by wild-type and vaccine strains of Yellow Fever virus. J Gen Virol 70: 557–564Google Scholar
  8. 8.
    Cavallaro KF, Werner MR Brinton MA (1990) Localization of N-glycosylation site(s) on West Nile virus (WNV) E protein. In: 8th International Congress of Virology, August 26–31, Berlin, Germany, p 34Google Scholar
  9. 9.
    Clewley JP, Bishop DH, Kang CY, Coffin J, Eichman ME, Schnitzlein WM, Shope RE (1977) Oligonucleotide fingerprints of RNA species obtained from rhabdoviruses belonging to the vesicular stomatitis virus subgroup. J Virol 23: 152–166Google Scholar
  10. 10.
    DeWachter R, Fiers W (1972) Preparative two-dimentional polyacrylamide gel electrophoresis of32P-labeled RNA. Anal Biochem 49: 184–197Google Scholar
  11. 11.
    Dunster LM, Gibson CA, Stephenson JR, Minor PD, Barret ADT (1990) Attenuation of virulence of flaviviruses following passage in HeLa cells. J Gen Virol 71: 601–607Google Scholar
  12. 12.
    Feinstein S, Akov Y, Lachmi B, Lehrer S, Rannon L, Katz D (1985) Determination of human IgG and IgM class antibodies to West Nile virus by enzyme-linked immunosorbent assay (ELISA). J Med Virol 17: 63–72Google Scholar
  13. 13.
    George G, Gourie-Devi M, Rao JA, Prasad SR, Pavri KM (1984) Isolation of West Nile virus from the brains of children who had died of encephalitis. Bull World Health Organ 62: 879–882Google Scholar
  14. 14.
    Gonzalez-Scarano F, Tyler KL (1987) Molecular pathogenesis of neurotropic viral infections. Ann Neurol 22: 565–574Google Scholar
  15. 14a.
    Gonzalez-Scarano F, Janssen RS, Najjar JA, Pobjecky N, Nathanson N (1985) An avirulent G1 glycoprotein variant of La Crosse Bunyavirus with defective fusion function. J Virol 54: 757–763Google Scholar
  16. 15.
    Green MC (1981) Genetic variants and strains of laboratory mouse. Fischer, StuttgartGoogle Scholar
  17. 16.
    Goldblum N, Sterk VV, Paderski B (1954) West Nile fever: the clinical features of the disease and the isolation of West Nile virus from the blood of nine human cases. Am J Hyg 59: 89–103Google Scholar
  18. 17.
    Halevy M, Lustig S, Akov Y (1986) Neuroinvasiveness and replication in murine macrophages of two West Nile virus (WNV) strains. Isr J Med Sci 22: 148Google Scholar
  19. 18.
    Hsieh P, Robbins PW (1984) Regulation of asparagine-linked oligosaccharide processing: oligosaccharide processing in Aedes albopictus mosquito cells. J Biol Chem 259: 2375–2382Google Scholar
  20. 19.
    Jacoby RO, Bhatt PN (1976) Genetic resistance to lethal Flavivirus encephalitis. I. Infection of congenic mice with Banzi virus. J Infect Dis 134: 158–165Google Scholar
  21. 20.
    Johnson RT (1982) Viral infection of the nervous system. Raven Press, New YorkGoogle Scholar
  22. 21.
    Kobiler D, Lustig, S, Gozes Y, Ben-Nathan D, Akov Y (1989) Sodium dodecylsulphate induces a breach in the blood-brain barrier and enables a West Nile virus variant to penetrate into mouse brain. Brain Res 496: 314–316Google Scholar
  23. 22.
    Lachmi B, Epstein N, Olshevsky U (1987) Characterization of five epitops on the envelope-protein of West Nile virus using monoclonal antibodies. In: Proceeding of the 7th International Congress of Virology, Edmonton, Alberta, Canada, p 107Google Scholar
  24. 23.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685Google Scholar
  25. 24.
    Lustig S, Halevy M, Akov Y, Shapira A (1985) Attenuation of West Nile (WN) virus in persistently WN-infected mosquito cell culture. Isr J Med Sci 21: 182Google Scholar
  26. 25.
    Lustig S, Danenberg HD, Kafri Y, Kobiler D, Ben-Nathan D (1992) Viral neuroinvasion and encephalitis induced by lipopolysaccharide and its mediators. J Exp Med 176: 707–712Google Scholar
  27. 26.
    Mandl CW, Guirakhoo F, Holzmann H, Heinz FX, Kunz C (1989) Antigenic structure of the Flavivirus envelope protein E at the molecular level, using tick-borne encephalitis virus as a model. J Virol 63: 564–571Google Scholar
  28. 27.
    Melnick JL, Paul JR, Riordan JT, Barnett VH, Goldblum N, Zabin E (1951) Isolation from human sera in Egypt of a virus apparently identical to West Nile virus. Proc Soc Exp Biol Med 77: 661–665Google Scholar
  29. 28.
    Minagawa H, Sakuma S, Mohri S, Mohri R, Watanabe T (1988) Herpes simplex virus type 1 infection in mice with severe combined immunodeficiency (SCID). Arch Virol 103: 73–82Google Scholar
  30. 29.
    Monath TP (1990) Flaviviruses. In: Fields BN, Knipe DM (eds) Virology, vol 1, 2nd edn. Raven Press, New York, pp 763–814Google Scholar
  31. 30.
    Monath TP, Cropp CB, Harrison AK (1993) Mode of entry of a neurotropic arbovirus into the central nervous system. Lab Invest 48: 399–410Google Scholar
  32. 31.
    Nowak T, Wengler G (1987) Analysis of disulfides present in the membrane proteins of the West Nile Flavivirus. Virology 156: 127–137Google Scholar
  33. 32.
    Poidinger M, Coelen RJ, Mackenzie JS (1991) Persistent infection of Vero cells by the flavivirus Murray Valley encephalitis virus. J Gen Virol 72: 573–578Google Scholar
  34. 33.
    Reed LJ, Muench H (1938) A simple method of establishing fifty percent end-points. Am J Hyg 270: 493–497Google Scholar
  35. 34.
    Strivastava AK, Aira Y, Mori C, Kobayashi Y, Igarashi A (1987) Antigenicity of Japanese encephalitis virus envelope protein V3 (E) and its cyanogen bromide cleaved fragments examined by monoclonal antibodies and Western blotting. Arch Virol 96: 97–107Google Scholar
  36. 35.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354Google Scholar
  37. 36.
    Wengler G, Wengler G, Gross HJ (1978) Studies on virus specific nucleic acid synthesized in vertebrate and mosquito cells infected with flaviviruses. Virology 89: 423–437Google Scholar
  38. 37.
    Wengler G, Castle E, Leidner U, Nowak T, Wengler G (1985) Sequence analysis of the membrane protein V3 of the Flavivirus West Nile virus and of its gene. Virology 147: 264–274Google Scholar
  39. 38.
    Wright PJ (1982) Envelope protein of the flavivirus Kunjin is apparently not glycosylated. J Gen Virol 59: 29–38Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • M. Halevy
    • 1
  • Y. Akov
    • 1
  • D. Ben-Nathan
    • 1
  • D. Kobiler
    • 1
  • B. Lachmi
    • 1
  • S. Lustig
    • 1
  1. 1.Department of VirologyIsrael Institute for Biological ResearchNess-ZionaIsrael

Personalised recommendations