Archives of Virology

, Volume 127, Issue 1–4, pp 153–168 | Cite as

Identification of epitopes associated with different biological activities on the glycoprotein of vesicular stomatitis virus by use of monoclonal antibodies

  • S. Nagata
  • Y. Okamoto
  • T. Inoue
  • Y. Ueno
  • T. Kurata
  • J. Chiba
Original Papers


Thirteen monoclonal antibodies (MAbs) to the glycoprotein (G) of vesicular stomatitis virus (VSV) serotype Indiana were prepared and examined for their effects on various biological activities of VSV, including in vitro infection, hemagglutination, adsorption to cells, and mediation of cell fusion. Competitive binding assays with these MAbs revealed the presence of at least seven distinct antigenic determinants (epitopes) on the G protein. In some cases, overlappings among epitopes to various degrees were observed as partial inhibition or binding enhancement. The MAbs to all the epitopes but one (epitopes 1–6) reacted with the denatured G protein in a Western immunoblot analysis. Four of the epitopes (epitopes 2, 4, 5, and 7) were involved in neutralization and two (epitopes 1 and 2) in hemagglutination inhibition. None of the MAbs inhibited the adsorption of radiolabeled VSV to BHK-21 cells; the MAbs to epitope 2 slightly enhanced the virus adsorption. All neutralization epitopes except epitope 2 (epitopes 4, 5, and 7) were associated with inhibition of VSV-mediated cell fusion. These results show a direct spatial relationship between the epitopes recognized by the MAbs and functional sites on G protein and further insights into the structure and function of G protein.


Monoclonal Antibody Binding Assay Immunoblot Analysis Spatial Relationship Stomatitis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bailey CA, Miller DK, Lenard J (1984) Effects of DEAE-dextran on infection and hemolysis by VSV. Evidence that nonspecific electrostatic interactions mediate effective binding of VSV to cells. Virology 133: 111–118Google Scholar
  2. 2.
    Bailey MJ, Mcleod DA, Kang C, Bishop DHL (1989) Glycosylation is not required for the fusion activity of the G protein of vesicular stomatitis virus in insect cells. Virology 169: 323–331Google Scholar
  3. 3.
    Bishop DHL, Repik P, Obijeski JF, Moore NF, Wagner RR (1975) Restitution of infectivity to spikeless vesicular stomatitis virus by solubilized viral components. J Virol 16: 75–84Google Scholar
  4. 4.
    Boere WAM, Harmsen T, Vinjé J, Benaissa-Trouw BJ, Kraaijeveld CA, Snippe H (1984) Identification of distinct antigenic determinants on Semliki Forest virus by using monoclonal antibodies with different antiviral activities. J Virol 52: 575–582Google Scholar
  5. 5.
    Bricker BJ, Snyder RM, Fox JW, Volk WA, Wagner RR (1987) Monoclonal antibodies to the glycoprotein of vesicular stomatitis virus (New Jersey serotype): a method for preliminary mapping of epitopes. Virology 161: 533–540Google Scholar
  6. 6.
    Cartwright B, Smale CJ, Brown F (1969) Surface structure of vesicular stomatitis virus. J Gen Virol 5: 1–10Google Scholar
  7. 7.
    Florkiewicz RZ, Rose JK (1984) A cell line expressing vesicular stomatitis virus glycoprotein fuses at low pH. Science 225: 721–723Google Scholar
  8. 8.
    Grigera PR, Mathieu ME, Wagner RR (1991) Effect of glycosylation on the conformational epitopes on the glycoprotein of vesicular stomatitis virus (New Jersey serotype). Virology 180: 1–9Google Scholar
  9. 9.
    Halonen PE, Murphy FA, Fields BN, Reese DR (1968) Hemagglutinin of rabies and some other bullet-shaped viruses. Proc Soc Exp Biol Med 127: 1037–1042Google Scholar
  10. 10.
    Hoekstra D (1990) Membrane fusion of envelope viruses: especially a matter of proteins. J Bioenerg Biomembr 22: 121–155Google Scholar
  11. 11.
    Keil W, Wagner RR (1989) Epitope mapping by deletion mutants and chimeras of two vesicular stomatitis virus glycoprotein genes expressed by a vaccinia virus vector. Virology 170: 392–407Google Scholar
  12. 12.
    Kelley JM, Emerson SU, Wagner RR (1972) The glycoprotein of vesicular stomatitis virus is the antigen that gives rise to and reacts with neutralizing antibody. J Virol 10: 1231–1235Google Scholar
  13. 13.
    Kingsford L, Ishizaka LD, Hill DW (1983) Biological activities of monoclonal antibodies reactive with antigenic sites mapped on the G1 glycoprotein of La Crosse virus. Virology 129: 443–445Google Scholar
  14. 14.
    LeFrancois L, Lyles DS (1982) The interaction of antibody with the major surface glycoprotein of vesicular stomatitis virus I. Analysis of neutralizing epitopes with monoclonal antibodies. Virology 121: 157–167Google Scholar
  15. 15.
    LeFrancois L, Lyles DS (1982) The interaction of antibody with the major surface glycoprotein of vesicular stomatitis virus II. Monoclonal antibodies to nonneutralizing and cross-reactive epitope of Indiana and New Jersey serotypes. Virology 121: 168–174Google Scholar
  16. 16.
    LeFrancois L, Lyles DS (1983) Antigenic determinants of vesicular stomatitis virus: analysis with antigenic variants. J Immunol 130: 394–398Google Scholar
  17. 17.
    Luo L, Li Y, Snyder RM, Wagner RR (1988) Point mutations in glycoprotein gene of vesicular stomatitis virus (New Jersey serotype) selected by resistance to neutralization by epitope-specific monoclonal antibodies. Virology 163: 341–348Google Scholar
  18. 18.
    Luo L, Li Y, Snyder RM, Wagner RR (1990) Spontaneous mutations leading to antigenic variations in the glycoproteins of vesicular stomatitis virus field isolates. Virology 174: 70–78Google Scholar
  19. 19.
    Mannen K, Ohuchi M, Mifune K (1982) pH-dependent hemolysis and cell fusion of rhabdoviruses. Microbiol Immunol 26: 1035–1043Google Scholar
  20. 20.
    Marsh M, Helenius A (1989) Virus entry into animal cells. Adv Virus Res 36: 107–151Google Scholar
  21. 21.
    Matlin KS, Reggio H, Helenius A, Simons K (1982) Pathway of vesicular stomatitis virus entry leading to infection. J Mol Biol 156: 609–631Google Scholar
  22. 22.
    Matlin K, Bainton DF, Pesonen M, Louvard D, Genty N, Simons K (1983) Trans-epithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence. J Cell Biol 97: 627–637Google Scholar
  23. 23.
    McSharry JJ, Ledda CA, Freiman HJ, Choppin PW (1978) Biological properties of VSV glycoprotein. II. Effects of the host cell and of the glycoprotein carbohydrate composition on hemagglutination. Virology 84: 183–188Google Scholar
  24. 24.
    Nagata S, Kurata T, Ueno Y, Chiba J (1991) Vesicular stomatitis virus-mediated cell fusion subsequent to virus adsorption at different pH values. Jpn J Med Sci Biol 44: 171–180Google Scholar
  25. 25.
    Nagata S, Yamamoto K, Ueno Y, Kurata T, Chiba J (1991) Production of monoclonal antibodies by the use of pH-dependent vesicular stomatitis virus-mediated cell fusion. Hybridoma 10: 317–322Google Scholar
  26. 26.
    Nagata S, Yamamoto K, Ueno Y, Kurata T, Chiba J (1991) Preferential generation of monoclonal IgG-producing hybridomas by use of vesicular stomatitis virus-mediated cell fusion. Hybridoma 10: 369–378Google Scholar
  27. 27.
    Ohnishi S (1988) Fusion of viral envelopes with cellular membrane. In: Bronner F, Düzgünes N (eds) Membrane fusion in fertilization, cellular transport, and viral infection. Academic Press, London, pp 257–296 (Current topics in membranes and transport, vol 32)Google Scholar
  28. 28.
    Oi VT, Herzenberg LA (1980) Immunoglobulin-producing hybrid cell lines. In: Mishell BB, Shiigi SM (eds) Selected methods in cellular immunology. Freeman, San Francisco, 351–372Google Scholar
  29. 29.
    Pal R, Barenholz Y, Wagner RR (1987) Vesicular stomatitis virus membrane proteins and their interactions with lipid bilayers. Biochim Biophys Acta 906: 175–193Google Scholar
  30. 30.
    Petri WA, Wagner RR (1979) Reconstitution into liposomes of the glycoprotein of vesicular stomatitis virus by detergent dialysis. J Biol Chem 254: 4313–4316Google Scholar
  31. 31.
    Routledge E, Stauber R, Pfleiderer M, Siddell SG (1991) Analysis of murine coronavirus surface glycoprotein functions by using monoclonal antibodies. J Virol 65: 254–262Google Scholar
  32. 32.
    Schlegel R, Wade M (1983) Neutralized vesicular stomatitis virus binds to host cells by a different “receptor”. Biochem Biophys Res Commun 114: 774–778Google Scholar
  33. 33.
    Talbot PJ, Salmi AA, Knobler RL, Buchmeier MJ (1984) Topographical mapping of epitopes on the glycoproteins of murine hepatitis virus-4 (strain JHM): correlation with biological activities. Virology 132: 250–260Google Scholar
  34. 34.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354Google Scholar
  35. 35.
    Vandepol SB, LeFrancois L, Holland JJ (1986) Sequences of the major antibody binding epitopes of the Indiana serotype of vesicular stomatitis virus. Virology 148: 312–325Google Scholar
  36. 36.
    Volk WA, Snyder RM, Benjamin DC, Wagner RR (1982) Monoclonal antibodies to the glycoprotein of vesicular stomatitis virus: comparative neutralizing activity. J Virol 42: 220–227Google Scholar
  37. 37.
    Wagner RR (1987) Rhabdovirus biology and infection: an overview. In: Wagner RR (ed) The rhabdovirus. Plenum, New York, pp 9–74Google Scholar
  38. 38.
    White J, Matlin K, Helenius A (1981) Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol 89: 674–679Google Scholar
  39. 39.
    Whitt MA, Zagouras P, Crise B, Rose JK (1990) A fusion-defective mutant of the vesicular stomatitis virus glycoprotein. J Virol 64: 4907–4913Google Scholar
  40. 40.
    Whitt MA, Rose JK (1991) Fatty acid acylation is not required for membrane fusion activity or glycoprotein assembly into VSV virions. Virology 185: 875–878Google Scholar
  41. 41.
    Wunner WH (1985) Growth, purification and titration of rhabdoviruses. In: Mahy BWJ (ed) Virology: a practical approach. IRL Press, Oxford, pp 79–93Google Scholar
  42. 42.
    Yamakawa Y, Chiba J (1988) High performance liquid chromatography of mouse monoclonal antibodies on spherical hydroxyapatite beads. J Liquid Chromatogr 11: 665–681Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • S. Nagata
    • 1
    • 2
  • Y. Okamoto
    • 1
    • 2
  • T. Inoue
    • 1
    • 2
  • Y. Ueno
    • 2
  • T. Kurata
    • 1
  • J. Chiba
    • 1
    • 3
  1. 1.Department of PathologyNational Institute of HealthTokyo
  2. 2.Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical SciencesScience University of TokyoTokyo
  3. 3.Laboratory of Immunology, Department of Biological Science and TechnologyScience University of TokyoNoda, ChibaJapan

Personalised recommendations