1887

Abstract

Summary

We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-l. We found that 8–20% gradient polyacrylamide gels provided no advantage over fixed 8·5% gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0·5% in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122–130) × 10-mol. wt region, a 75 × 10-mol. wt protein, the gD region of approximately (56–64) × 10-mol. wt and two non-glycosylated bands at mol. wts(10) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD > gB/gC > (42/44) × 10≫P75≫VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-25-1-49
1988-01-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/25/1/medmicro-25-1-49.html?itemId=/content/journal/jmm/10.1099/00222615-25-1-49&mimeType=html&fmt=ahah

References

  1. Ashley R L, Corey L. 1984; Effect of acyclovir treatment of primary genital herpes on the antibody response to herpes simplex virus. Journal of Clinical Investigation 73:681–688
    [Google Scholar]
  2. Baucke R B, Spear P G. 1979; Membrane proteins specified by herpes simplex viruses. V. Identification of an Fc-binding glycoprotein. Journal of Virology 32:779–789
    [Google Scholar]
  3. Braun D K, Pereira L, Norrild B, Roizman B. 1983; Application of denatured, electrophoretically separated, and immobilized lysates of herpes simplex virus-infected cells for detection of monoclonal antibodies and for studies of the properties of viral proteins. Journal of Virology 46:103–112
    [Google Scholar]
  4. Cohen G H, Long D, Eisenberg R J. 1980; Synthesis and processing of glycoproteins gD and gC of herpes simplex virus type 1. Journal of Virology 36:429–439
    [Google Scholar]
  5. Eberle R, Mou S-W. 1983; Relative titres of antibodies to individual polypeptide antigens of herpes simplex virus type 1 in human sera. Journal of Infectious Diseases 148:436–444
    [Google Scholar]
  6. Eberle R, Mou S-W, Zaia J A. 1984; Polypeptide specificity of the early antibody response following primary and recurrent genital herpes simplex virus type 2 infections. Journal of General Virology 65:1839–1843
    [Google Scholar]
  7. Eberle R, Mou S-W, Zaia J A. 1985; The immune response to herpes simplex virus: Comparison of the specificity and relative titres of serum antibodies directed against viral polypeptides following primary herpes simplex virus type 1 infections. Journal of Medical Virology 16:147–162
    [Google Scholar]
  8. Gershoni J M, Palade G E. 1983; Protein blotting: principles and applications. Analytical Biochemistry 131:1–15
    [Google Scholar]
  9. Gilman S C, Docherty J J, Rawls W E. 1981; Antibody responses in humans to individual proteins of herpes simplex viruses. Infection and Immunity 34:880–887
    [Google Scholar]
  10. Heilman C J, Zweig M, Hampar B. 1981; Herpes simplex virus type 1 and 2 intracellular p40: Type-specific and cross-reactive antigenic determinants on peptides generated by partial proteolysis. Journal of Virology 40:508–515
    [Google Scholar]
  11. Laemmli U K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  12. Lee G T-Y, Pogue-Geile K L, Pereira L, Spear P G. 1982; Expression of herpes simplex virus glycoprotein C from a DNA fragment inserted into the thymidine kinase gene of this virus. Proceedings of the National Academy of Sciences of the USA 79:6612–6616
    [Google Scholar]
  13. Lehtinen M, Koivisto V, Lehtinen T, Paavonen J, Leinikki P. 1985; Immunoblotting and enzyme-linked immunosorbent assay analysis of serological responses in patients infected with herpes simplex virus type 1 and type 2. Intervirology 24:18–25
    [Google Scholar]
  14. Mann D R, Hilty M D. 1982; Antibody response to herpes simplex virus type 1 polypeptides and glycoproteins in primary and recurrent infection. Pediatrie Research 16:176–180
    [Google Scholar]
  15. Norrild B. 1980; Immunochemistry of herpes simplex virus glycoproteins. Current Topics in Microbiology and Immunology 90:68–106
    [Google Scholar]
  16. Norrild B. 1985; Humoral response to herpes simplex virus infections. Roizman B, Lopez C. The Herpes-viruses 4 Plenum Press; New York, NY:69–86
    [Google Scholar]
  17. Norrild B, Pedersen B, Roizman B. 1981; Immunological reactivity of herpes simplex virus 1 and 2 polypeptides electrophoretically separated and transferred to diazoben-zyloxymethyl paper. Infection and Immunity 31:660–667
    [Google Scholar]
  18. Pauli G, Ludwig H. 1977; Immunoprecipitation of herpes simplex virus type 1 antigens with different antisera and human cerebrospinal fluids. Archives of Virology 53:139–155
    [Google Scholar]
  19. Preston V G, Coates J A V, Rixon F J. 1983; Identification and characterization of a herpes simplex virus gene product required for encapsidation of virus DNA. Journal of Virology 45:1056–1064
    [Google Scholar]
  20. Ratner J J, Smith K O. 1980; Serum antibodies to herpes simplex virus type 1 during active oral herpes infection. Infection and Immunity 27:113–117
    [Google Scholar]
  21. Towbin H, Staehelin T, Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : Procedure and some applications. Proceedings of the National Academy of Sciences of the USA 76:4350–4354
    [Google Scholar]
  22. Vestergaard B F. 1979; Quantitative immunoelectrophoretic analysis of human antibodies against herpes simplex virus antigens. Infection and Immunity 23:553–558
    [Google Scholar]
  23. Wenske E A, Bratton M W, Courtney R J. 1982; Endo-β-N-acetylglucosaminidase H sensitivity of precursors to herpes simplex virus type 1 glycoproteins gB and gC. Journal of Virology 44:241–248
    [Google Scholar]
  24. Zweerink H J, Corey L. 1982; Virus-specific antibodies in sera from patients with genital herpes simplex virus infection. Infection and Immunity 37:413–421
    [Google Scholar]
  25. Zweerink H J, Stanton L W. 1981; Immune response to herpes simplex virus infections: Virus-specific antibodies in sera from patients with recurrent facial infections. Infection and Immunity 31:624–630
    [Google Scholar]
  26. Zweig M, Heilman C J, Rabin H, Hampar B. 1980; Shared antigenic determinants between two distinct classes of proteins in cells infected with herpes simplex virus. Journal of Virology 35:644–652
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-25-1-49
Loading
/content/journal/jmm/10.1099/00222615-25-1-49
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error