1887

Abstract

Hybrid cell lines have been derived which produce monoclonal antibodies reacting with outer membrane protein I from strain P9. The antibodies obtained showed variable reactivity with other strains but one antibody recognized an epitope present on all of the strains tested which expressed the protease sensitive protein IB. Purified IgG labelled with I was used in competitive radioimmunoassays with unlabelled antibody to investigate the spacial distribution of the epitopes recognized. Each pair of antibodies showed some degree of inhibition. The relative magnitude of inhibition suggested that the conserved epitope lies within a variable region containing other epitopes which determine the antigenic specificity of the protein. Western blotting of peptides derived by proteolytic digestion of protein IB revealed that the conserved epitope is located close to the chymotrypsin cleavage site within a 7000 surface exposed region of the molecule.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-132-6-1611
1986-06-01
2021-10-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/132/6/mic-132-6-1611.html?itemId=/content/journal/micro/10.1099/00221287-132-6-1611&mimeType=html&fmt=ahah

References

  1. Barrera O., Swanson J. 1984; Proteins IA and IB exhibit different surface exposures and orientations in the outer membranes of Neisseria gotiorrhoeae. . Infection and Immunity 44:565–568
    [Google Scholar]
  2. Blake M. S., Gotschlich E. C. 1983; Gonococcal membrane proteins: speculation on their role in pathogenesis.. Progress in Allergy 33:298–313
    [Google Scholar]
  3. Blake M. S., Gotschlich E. C., Swanson J. 1981; Effects of proteolytic enzymes on the outer membrane proteins of Neisseria gonorrhoeae.. Infection and Immunity 33:212–222
    [Google Scholar]
  4. DIAZ J.-L., Heckels J. E. 1982; Antigenic variation of outer membrane protein II in colonial variants of Neisseria gonorrhoeae P9. . Journal of General Microbiology 128:585–591
    [Google Scholar]
  5. Diaz J.-L., Virji M., Heckels J. E. 1984; Structural and antigenic differences between two types of meningococcal pili. . FEMS Microbiology Letters 21:181–184
    [Google Scholar]
  6. Douglas J. T., Lee M. D., Nikaido H. 1981; Protein I of Neisseria gonorrhoeae outer membrane is a porin. . FEMS Microbiology Letters 12:305–309
    [Google Scholar]
  7. Frankel M. E., Gerhard W. 1979; The rapid determination of binding constants for antiviral antibodies by a radioimmunoassay. An analysis of the interaction between hybridoma proteins and influenza virus. . Molecular Immunology 16:101–106
    [Google Scholar]
  8. Hashimoto F., Horigome T., Kanabayashi M., Yoshida K., Sugano H. 1983; An improved method for separation of low-molecular-weight polypeptides by electrophoresis in sodium dodecyl sulfate-polyacrylamide gel. . Analytical Biochemistry 129:192–199
    [Google Scholar]
  9. Heckels J. E. 1977; The surface properties of Neisseria gonorrhoeae: isolation of the major components of the outer membrane. . Journal of General Microbiology 99:333–341
    [Google Scholar]
  10. Heckels J. E. 1981; Structural comparison of Neisseria gonorrhoeae outer membrane proteins. . Journal of Bacteriology 145:736–742
    [Google Scholar]
  11. Heckels J. E. 1986; Gonococcal antigenic variation and pathogenesis. . In Antigenic Variation in the Course of Infectious Diseases: a Survival Strategy for Pathogenic Microorganisms. editor Birkbeck H., Penn C. W. . Cambridge: Cambridge University Press; in the Press
    [Google Scholar]
  12. Heckels J. E., Virji M. 1985; Monoclonal antibodies against gonococcal pili: uses in analysis of gonococcal immunochemistry and virulence.. In Monoclonal Antibodies Against Bacteria vol. l pp. 1–35 editor Macario A. J. L., de Macario E. C. . London: Academic Press;
    [Google Scholar]
  13. Knapp J. S., Tam M. R., Nowinski R. C., Holmes K. K., Sandstrom E. G. 1984; Serological classification of Neisseria gonorrhoeae with use of monoclonal antibodies to gonococcal outer membrane protein I. . Journal of Infectious Diseases 150:44–48
    [Google Scholar]
  14. Lambden P. R., Heckels J. E. 1982; Synthesis of immunogenic oligosaccharide-protein conjugates from the lipopolysaccharide of Neisseria gonorrhoeae P9. . Journal of Immunological Methods 48:233–240
    [Google Scholar]
  15. Lambden P. R., Heckels J. E., James L. T., Watt P. J. 1979; Variations in surface protein composition associated with virulence properties in opacity types of Neisseria gonorrhoeae. . Journal of General Microbiology 114:305–312
    [Google Scholar]
  16. Lambden P. R., Heckels J. E., McBride H., Watt P. J. 1981 a; The identification and isolation of novel pilus types produded by variants of N. gonorrhoeae P9 following selection in vivo.. FEMS Microbiology Letters 10:339–341
    [Google Scholar]
  17. Lambden P. R., Robertson J. N., Watt P. J. 1981b; The preparation and properties of a and ft pili from variants of Neisseria gonorrhoeae P9. . Journal of General Microbiology 124:109–117
    [Google Scholar]
  18. Lynch E. C., Blake M. S., Gotschlich E. C., Mauro A. 1984; Studies of porins: spontaneously transferred from whole cells and reconstituted from purified proteins of Neisseria gonorrhoeae and Neisseria meningitidis.. Biophysical Journal 45:104–107
    [Google Scholar]
  19. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. 1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. . Science 211:1437–1438
    [Google Scholar]
  20. Sandstrom E. G., Danielsson D. 1980; Serology of Neisseria gonorrhoeae. Classification by co-agglu- tination. . Acta pathologica et microbiologica scandina- vica B 33:27–38
    [Google Scholar]
  21. Sandstrom E. G., Chen K. C. S., Buchanan T. M. 1982; Serology of Neisseria gonorrhoeae: co-agglu- tination serogroups WI and WII/III correspond to different outer membrane protein molecules.. Infection and Immunity 38:462–470
    [Google Scholar]
  22. Swanson J. 1979; Studies on gonococcus infection. XVIII. 125I-labeled peptide mapping of the major protein of the gonococcal cell wall outer membrane.. Infection and Immunity 23:799–810
    [Google Scholar]
  23. Swanson J., Sparks E., Young D., King G. 1975; Studies on gonococcus infection. X. Pili and leukocyte association factor as mediators of interactions between gonococci and eukaryotic cells in vitro.. Infection and Immunity 11:1352–1361
    [Google Scholar]
  24. Towbin H., Stachelin T., Gordon J. 1979; Electrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications.. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4359
    [Google Scholar]
  25. Virji M., Everson J. S. 1981; Comparative virulence of opacity variants of Neisseria gonorrhoeae strain P9. . Infection and Immunity 31:965–970
    [Google Scholar]
  26. Virji M., Heckels J. E. 1983; Antigenic crossreactivity of Neisseria pili: investigations with type- and species-specific monoclonal antibodies. . Journal of General Microbiology 129:2761–2768
    [Google Scholar]
  27. Virji M., Heckels J. E. 1986; The effect of protein II and pili on the interaction of Neisseria gonorrhoeae with human polymorphonuclear leucocytes. . Journal of General Microbiology 132:503–512
    [Google Scholar]
  28. Virji M., Heckels J. E., Watt P. J. 1983; Monoclonal antibodies to gonococcal pili: studies on antigenic determinants on pili from variants of strain P9. . Journal of General Microbiology 129:1965–1973
    [Google Scholar]
  29. Virji M., Zak K., Heckels J. E. 1985; Use of monoclonal antibodies to detect common protein antigens present in outer membranes of Neisseria species. . In The Pathogenic Neisseriae pp. 276–280 editor Schoolnik G. K. Washington, DC:: American Society for Microbiology;
    [Google Scholar]
  30. Virji M., Zak K., Heckels J. E. 1986; Monoclonal antibodies to gonococcal outer membrane protein IB: use in investigation of the potential protective effect of antibodies directed against conserved and type-specific epitopes. . Journal of General Microbiology 1 32:1621–1629
    [Google Scholar]
  31. Wang S.-P., Holmes K. K., Knapp J. S., Ott S., Kyzer D. D. 1977; Immunologic classification of Neisseria gonorrhoeae with micro-immunofluorescence. . Journal of Immunology 119:795–803
    [Google Scholar]
  32. Zak K., Diaz J., -L., Jackson D., Heckels J. E. 1984; Antigenic variation during infection with Neisseria gonorrhoeae: detection of antibodies to surface proteins in sera of patients with gonorrhea. . Journal of Infectious Diseases 149:166–173
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-132-6-1611
Loading
/content/journal/micro/10.1099/00221287-132-6-1611
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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