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Volume 71, Issue 12

Epitope Analysis of the Envelope and Non-structural Glycoproteins of Murray Valley Encephalitis Virus Free

Abstract

Previous studies have shown that antibodies produced against strategic flavivirus epitopes play an important role in recovery and immunity. Definition of the conformation and location of these epitopes and the degree of their conservation among flaviviruses is important to understanding the humoral response to flavivirus infection. In this study we have examined epitopes recognized by 14 monoclonal antibodies (MAbs) produced to the envelope (E) and nonstructural (NS1) proteins of Murray Valley encephalitis virus (MVE). These antibodies were analysed for specificity, neutralization, haemagglutination inhibition (HI) and competitive binding. We have identified six distinct epitopes on the E protein which are located in four non-overlapping domains. MAbs to epitopes in one domain neutralized virus, were specific for MVE and Japanese encephalitis virus, and reacted with epitopes resistant to reduction. Two other E domains, one specific to MVE and the other shared by all flaviviruses, also contained neutralization sites and were stabilized by disulphide bonds. The fourth domain on E was conserved among the flaviviruses, sensitive to SDS denaturation and did not induce neutralizing antibody. Studies with MVE NS1 MAbs revealed that they were mostly type-specific, unreactive with conserved epitopes, and unreactive in HI and neutralization tests. The six epitopes identified on NS1 did not overlap and represent antigenic domains either resistant or sensitive to reduction. Immunoblotting of viral proteins in MVE-infected C6/36 cells revealed two distinct forms of NS1 and high proteins of 97K and 108K that represented disulphide-linked heterodimers of E and NS1.

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  • Accepted:
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© Society for General Microbiology 1990
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1990-12-01
2024-04-23
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References

  1. Arumugham R. G., Hildreth S. W., Paradiso P. R. 1989; Interprotein disulfide bonding between F and G glycoproteins of human respiratory syncytial virus. Archives of Virology 105:65–79
     [Google Scholar]
  2. Burke D. S., Nisalak A., Gentry M. K. 1987; Detection of flavivirus antibodies in human serum by epitope-blocking immunoassay. Journal of Medical Virology 23:165–173
     [Google Scholar]
  3. Castle E., Nowak T., Leidner U., Wengler G., Wengler G. 1985; Sequence analysis of the viral core protein and the membrane-associated proteins VI and NV2 of the flavivirus West Nile virus and the genome sequence of these proteins. Virology 145:227–236
     [Google Scholar]
  4. Coelen R. J., Mackenzie J. S. 1988; Genetic variation of Murray Valley encephalitis virus. Journal of General Virology 69:1903–1912
     [Google Scholar]
  5. Coia G., Parker M. D., Speight G., Byrne M. E., Westaway E. G. 1988; Nucleotide and complete amino acid sequences of Kunjin virus: definitive gene order and characteristics of the virus-specified proteins. Journal of General Virology 69:1–21
     [Google Scholar]
  6. Cook I. F.editor 1987; Epidemiological features of Australian encephalitis. Communicable Diseases Intelligence (Australia) 87/2:12–14
     [Google Scholar]
  7. Cook I. F.editor 1988; Australian encephalitis in the Northern Territory-4 case reports. Communicable Diseases Intelligence (Australia,) 88/12:2–6
     [Google Scholar]
  8. Doherty R. 1977; Arthropod-borne viruses in Australia. Australian Journal of Experimental Biology and Medical Sciences 55:103–130
     [Google Scholar]
  9. Galfré G., Hove S. C., Milstein C., Butcher G. W., Howard J. C. 1977; Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature; London: 266550–552
     [Google Scholar]
  10. Gibson C. A., Schlesinger J. J., Barrett A. D. T. 1988; Prospects for a virus non-structural protein as a subunit vaccine. Vaccine 6:7–9
     [Google Scholar]
  11. Goding J. W. 1986; Immunofluorescence. In Monoclonal Antibodies: Principles and Practice, 2nd edn.. pp 241–274 London: Academic Press;
     [Google Scholar]
  12. Gould E. A., Buckley A., Cammack N., Barrett A. D. T., Clegg J. C. S., Ishak R., Varma M. G. R. 1985; Examination of the immunological relationships between flaviviruses using yellow fever virus monoclonal antibodies. Journal of General Virology 66:1369–1382
     [Google Scholar]
  13. Guirakhoo F., Heinz F. X., Kunz C. 1989; Epitope model of tick-borne encephalitis virus envelope glycoprotein E: analysis of structural properties, role of carbohydrate side chain, and conformational changes occuring at acidic pH. Virology 169:90–99
     [Google Scholar]
  14. Hall R. A. 1989 Epitope analysis of Australian flaviviruses and applications in diagnosis and surveillance Ph.D thesis James Cook University of North Queens1And:
     [Google Scholar]
  15. Hall R. A., Kay B. H., Burgess G. W. 1987; An enzyme immunoassay to detect Australian flaviviruses and identify the encephalitic subgroup using monoclonal antibodies. Immunology and Cell Biology 65:103–110
     [Google Scholar]
  16. Hall R. A., Burgess G. W., Kay B. H. 1988; Type-specific monoclonal antibodies produced to proteins of Murray Valley encephalitis virus. Immunology and Cell Biology 66:51–56
     [Google Scholar]
  17. Hawkes R. A., Roehrig J. T., Hunt A. R., Moore G. A. 1988; Antigenic structure of the Murray Valley encephalitis virus E glycoprotein. Journal of General Virology 69:1105–1109
     [Google Scholar]
  18. Heinz F. X. 1986; Epitope mapping of flavivirus glycoproteins. Advances in Virus Research 31:103–168
     [Google Scholar]
  19. Heinz F. X., Berger R., Tuma W., Kunz C. 1983; A topological and functional model of epitopes on the structural glycoprotein of tick-borne encephalitis virus defined by monoclonal antibodies. Virology 126:525–537
     [Google Scholar]
  20. Henchal E. A., Mccown J. M., Burke D. S., Seguin M. C., Brandt W. E. 1985; Epitopic analysis of antigenic determinants on the surface of dengue-2 virions using monoclonal antibodies. American Journal of Tropical Medicine and Hygiene 34:162–169
     [Google Scholar]
  21. Henchal E. A., Henchal L. S., Thaisomboonsuk B. K. 1987; Topological mapping of unique epitopes on the dengue-2 virus NS1 protein using monoclonal antibodies. Journal of General Virology 68:845–851
     [Google Scholar]
  22. Henchal E. A., Henchal L. S., Schlesinger J. J. 1988; Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue 2 virus. Journal of General Virology 69:2101–2107
     [Google Scholar]
  23. Kaufman B. M., Summers P. L., Dubois D. R., Cohen W. H., Gentry M. K., Timchak R. L., Burke D. S., Eckels K. H. 1989; Monoclonal antibodies for dengue virus prM glycoprotein protect mice against lethal dengue infection. American Journal of Tropical Medicine and Hygiene 41:567–580
     [Google Scholar]
  24. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
     [Google Scholar]
  25. Mandl C. W., Guirakhoo F., Holzmann H., Heinz F. X., Kunz C. 1989; Antigenic structure of the flavivirus envelope protein E at the molecular level using tick-borne encephalitis virus as a model. Journal of Virology 63:564–571
     [Google Scholar]
  26. Mason P. W. 1989; Maturation of Japanese encephalitis virus glycoproteins produced by infected mammalian and mosquito cells. Virology 169:354–364
     [Google Scholar]
  27. Mason P. W., Mcada P. C., Dalrymple J. M., Fournier M. J., Mason T. L. 1987; Expression of Japanese encephalitis virus antigens in Escherichia coli. Virology 158:361–372
     [Google Scholar]
  28. Monath T. P. 1986; Pathobiology of the flaviviruses. In The Togaviridae and Flaviviridae pp 375–440 Schlesinger S., Schlesinger M. J. Edited by New York: Academic Press;
     [Google Scholar]
  29. Nowak T., Wengler G. 1987; Analysis of disulfides present in the membrane proteins of the West Nile flavivirus. Virology 156:127–137
     [Google Scholar]
  30. Rice C. M., Lenches E. M., Eddy S. R., Shin S. J., Sheets R., Strauss J. H. 1985; Nucleotide sequence of yellow fever virus; implications for flavivirus gene expression and evolution. Science 229:726–733
     [Google Scholar]
  31. Roehrig J. T., Hunt A. R., Johnson A. J., Hawkes R. A. 1989; Synthetic peptides derived from the deduced amino acid sequence of the E-glycoprotein of Murray Valley encephalitis virus elicit antiviral antibody. Virology 171:49–60
     [Google Scholar]
  32. Schlesinger J. J., Brandriss M. W., Monath T. P. 1983; Monoclonal antibodies distinguish between wild type vaccine strains of yellow fever virus by neutralisation, haemagglutination inhibition and immune precipitation of the virus envelope protein. Virology 125:8–17
     [Google Scholar]
  33. Schlesinger J. J., Brandriss M. W., Walsh E. E. 1985; Protection against 17D yellow fever encephalitis in mice by passive transfer of monoclonal antibodies to the nonstructural glycoprotein gp48 and by active immunisation with gp48. Journal of Immunology 135:2805–2809
     [Google Scholar]
  34. Schlesinger J. J., Brandriss M. W., Cropp C. B., Monath T. P. 1986; Protection against yellow fever in monkeys by immunisation with yellow fever virus nonstructural protein NS1. Journal of Virology 60:1153–1155
     [Google Scholar]
  35. Stanley N. F. 1982; Human arbovirus infections in Australia. Arbovirus Research in Australia 3:216–226
     [Google Scholar]
  36. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocelulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences U.S.A.: 764350–4354
     [Google Scholar]
  37. Trent D. W. 1977; Antigenic characterization of flavivirus structural proteins separated by isoelectric focusing. Journal of Virology 22:608–618
     [Google Scholar]
  38. Westaway G. 1987; Flavivirus replication strategy. Advances in Virus Research 33:45–90
     [Google Scholar]
  39. Winkler G., Randolph V. B., Cleaves G. R., Ryan T. E., Stollar V. 1988; Evidence that the mature form of the flavivirus nonstructural protein NS1 is a dimer. Virology 162:187–196
     [Google Scholar]
  40. Young P. R., Falconar A. K. I. 1989; Non-structural proteins as virus vaccines. Arbovirus Research in Australia 5:62–67
     [Google Scholar]
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Epitope Analysis of the Envelope and Non-structural Glycoproteins of Murray Valley Encephalitis Virus
J. Gen. Virol. 71, 2923 (1990); https://doi.org/10.1099/0022-1317-71-12-2923
/content/journal/jgv/10.1099/0022-1317-71-12-2923
/content/journal/jgv/10.1099/0022-1317-71-12-2923
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