1887

Abstract

The neisserial Lip antigen is a conserved antigen associated with the pathogenic species, and is composed of multiple repeats of a consensus pentapeptide. A series of monoclonal antibodies reacting with meningococcal Lip antigen were subjected to epitope mapping, using solid-phase synthetic peptides based on the consensus repeat sequence. The antibodies were found to recognize different continuous epitopes based on the consensus sequence. One monoclonal antibody was utilized in affinity chromatography to obtain purified Lip antigen and the antigen was used for immunization of mice. The resulting antisera did not recognize Lip antigen on Western blots but reacted specifically with Lip antigen in immune precipitation experiments, indicating that the predominant polyclonal immune response was directed against conformational epitopes. Despite the diversity of both continuous and conformational epitopes recognized by the antibodies produced, none of the antibodies demonstrated the ability to promote complement-mediated bactericidal activity. Thus despite its initial apparent promise as a potential vaccine candidate the case for the inclusion of Lip antigen in vaccine formulation cannot be supported at present.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-138-11-2321
1992-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/138/11/mic-138-11-2321.html?itemId=/content/journal/micro/10.1099/00221287-138-11-2321&mimeType=html&fmt=ahah

References

  1. Baehr W., Gotschlich E. C., Hitchcock P. J. 1989; The virulence-associated gonococcal H.8 gene encodes 14 tandemly repeated pentapeptides. Molecular Microbiology 3:49–55
    [Google Scholar]
  2. Bhatiacharjee A. K., Moran E. E., Ray J. S., Zollinger W. D. 1988; Purification and characterisation of the H.8 antigen from group B Neisseria meningitidis. Infection and Immunity 56:773–778
    [Google Scholar]
  3. Batiacharjee A. K., Moran E. E., Zollinger W. D. 1990; Antibodies to meningococcal H.8 (Lip) antigen fail to show bactericidal activity. Canadian Journal of Microbiology 36:117–122
    [Google Scholar]
  4. Black J. R., Cannon J. G. 1984; Monoclonal antibody (McAb) to common pathogenic Neisseria antigen (H8Ag) protects against meningococcemia in mice. Abstracts of the 24th Interstate Conference on Antimicrobial Agents and Chemotherapy, abstract 96. Washington DC: American Society for Microbiology;
    [Google Scholar]
  5. Black J. R., Black W. J., Cannon J. G. 1985; Neisserial antigen H.8 is immunogenic in patients with disseminated gonococcal and meningococcal infections. Journal of Infectious Diseases 151:650–657
    [Google Scholar]
  6. Bolton A. E., Hunter W. M. 1973; The labelling of proteins to high specific radioactivities by conjugation to a 1251-containing acylating agent. Application to the radioimmunoassay. Biochemical Journal 133:529–539
    [Google Scholar]
  7. Cannon J. G., Black W. J., Nachamkin I., Stewart P.W. 1984; Monoclonal antibody that recognises an outer membrane antigen common to the pathogenic Neisseria species but not to most nonpathogenic Neisseria species. Infection and Immunity 43:994–999
    [Google Scholar]
  8. 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]
  9. Fairbanks G., Steck T. L., Wallach D. F. H. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617
    [Google Scholar]
  10. Frasch C. E., Zollinger W. D., Poolman J. T. 1985; Serotype antigens of Neisseria meningitidis and a proposed scheme for designation of serotypes. Reviews of Infectious Diseases 1:504–510
    [Google Scholar]
  11. Gold R., Lepow M. L., Goldschneider I., Draper T. F., Gotschlich E. C. 1979; Kinetics of antibody production to group A and group C meningococcal polysaccharide vaccines administered during the first six years of life: prospects for routine immunization of infants and children. Journal of Infectious Diseases 140:690–697
    [Google Scholar]
  12. Goldschneider I., Gotschlich E. C., Artenstein M.S. 1969; Human immunity to the meningococcus. II. Development of natural immunity. Journal of Experimental Medicine 129:1327–1348
    [Google Scholar]
  13. Heckels J.E. 1981; Structural comparison of Neisseria gonorrhoeae outer membrane proteins. Journal of Bacteriology 145:736–742
    [Google Scholar]
  14. Hitchcock P. J. 1989; Unified nomenclature for pathogenic Neisseria species. Clinical Microbiology Reviews 2:564–565
    [Google Scholar]
  15. Hitchcock P. J., Brown T. M. 1983; Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silverstained polyacrylamide gels. Journal of Bacteriology 154:269–277
    [Google Scholar]
  16. Hitchcock P. J., Hayes S. F., Mayer L. W., Shafer W. M., Tessier S. L. 1985; Analyses of gonococcal H.8 antigen. Surface location, inter- and intra-strain electrophoretic heterogeneity, and unusual two-dimensional electrophoretic characteristics. Journal of Experimental Medicine 162:2017–2034
    [Google Scholar]
  17. Kawula T. H., Spinola S. M., Klapper D. G., Cannon J. G. 1987; Localisation of a conserved epitope and an azurin-like domain in the H.8 protein of pathogenic Neisseria. Molecular Microbiology 1:179–185
    [Google Scholar]
  18. Khyse-Andersen J. 1984; Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. Journal of Biochemical and Biophysical Methods 10:203–209
    [Google Scholar]
  19. Mcguinness B. T., Clarke I. N., Lambden P.R., Barlow A. K., Poolman J. T., Jones D. M., Heckels J. E. 1991; Point mutation in meningococcal porA gene associated with increased endemic disease. Lancet 337:514–517
    [Google Scholar]
  20. Mclachlan A. D. 1978; The double helix coiled coil structure of murein lipoprotein from Escherichia coli. Journal of Molecular Biology 122:493–506
    [Google Scholar]
  21. Munkley A., Tinsley C. R., Virji M., Heckels J. E. 1991; Blocking of bactericidal killing of Neisseria meningitidis by antibodies directed against class 4 outer membrane protein. Microbial Pathogenesis 11:447–452
    [Google Scholar]
  22. Peltola H. 1983; Meningococcal disease: still with us. Reviews of Infectious Diseases 5:71–91
    [Google Scholar]
  23. Peltola H., Safary A., Kayhty H., Karanko V., Andre F. E. 1985; Evaluation of two tetravalent (ACYW 135) meningococcal vaccines in infants and small children: a clinical study comparing immunogenicity of O-acetyl-negative and O-acetyl-positive group C polysaccharides. Pediatrics 16:91–96
    [Google Scholar]
  24. Phillips G. N., Flicker P. F., Cohen C., Manjula B. N., Fischetti V. A. 1981; Streptococcal M protein: ex-helical coiledcoil structure and arrangement on the cell surface. Proceedings of the National Academy of Sciences of the United States of America 784689–4693
    [Google Scholar]
  25. Poolman J. T., De Marie S., Zanen H. C. 1980; Variability of low-molecular-weight, heat-modifiable outer membrane proteins of Neisseria meningitidis. Infection and Immunity 30:642–648
    [Google Scholar]
  26. Poolman J. T., Hopman C. T. P., Zanen H. C. 1983; Immunogenicity of meningococcal antigens as detected in patient sera. Infection and Immunity 40:398–406
    [Google Scholar]
  27. Reingold A. L., Broome C. V., Hightower A. W., Ajelo G. W., Bolan G. A., Adamsbaum C., Jones E. E., Phillips C., Tiendrebeogo H., Yada A. 1985; Age-specific differences in duration of clinical protection after vaccination with meningococcal polysaccharide A vaccine. Lancet 2:114–118
    [Google Scholar]
  28. Robinson E. N., Mcgee Z. A., Buchanan T. M., Blake M. S., Hitchcock P. J. 1987; Probing the surface of Neisseria gonorrhoeae: simultaneous localisation of PI and H.8 antigens. Infection and Immunity 55:1190–1197
    [Google Scholar]
  29. Schweinle J.E., Hitchcock P. J., Tenner A. J., Hammer C.H., Frank M. M., Joiner K. A. 1989; Interaction of Neisseria gonorrhoeae with classical complement components, Cl-inhibitor, and a monoclonal antibody directed against the neisserial H.8 antigen. Journal of Clinical Investigation 83:397–403
    [Google Scholar]
  30. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. 1985; Measurement of protein using bicinchoninic acid. Analytical Biochemistry 150:76–85
    [Google Scholar]
  31. Strittmatter W., Hitchcock P. J. 1986; Isolation and preliminary biochemical characterization of the gonococcal H.8 antigen. Journal of Experimental Medicine 164:2038–2048
    [Google Scholar]
  32. Tinsley C.R., Heckels J.E. 1986; Variation in the expression of pili and outer membrane protein by Neisseria meningitidis during the course of meningococcal infection. Journal of General Microbiology 132:2483–2490
    [Google Scholar]
  33. Virji M., Heckels J.E. 1988; Nonbactericidal antibodies against Neisseria gonorrhoeae: evaluation of their blocking effect on bactericidal antibodies directed against outer membrane antigens. Journal of General Microbiology 134:2703–2711
    [Google Scholar]
  34. Virji M., Heckels J. E. 1989; Location of a blocking epitope on outer membrane protein III of Neisseria gonorrhoeae by synthetic peptide analysis. Journal of General Microbiology 135:1895–1899
    [Google Scholar]
  35. 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: Proceedings of the Fourth International Symposium, pp. 276–280 Edited by Schoolnik G. K. Washington DC: American Society for Microbiology;
    [Google Scholar]
  36. Virji M., Fletcher J. N., Zak K., Heckels J. E. 1987; The potential protective effect of monoclonal antibodies to gonococcal outer membrane protein IA. Journal of General Microbiology 133:2639–2646
    [Google Scholar]
  37. Wedege E., Froholm L.O. 1986; Human antibody response to a group B serotype 2A meningococcal vaccine determined by immunoblotting. Infection and Immunity 51:571–578
    [Google Scholar]
  38. Woods J.P., Black J. R., Barrit D.S., Connel T. D., Cannon J. G. 1987; Resistance to meningococcaemia apparently conferred by anti-H.8 monoclonal antibody is due to contaminating endotoxin and not to specific immunoprotection. Infection and Immunity 55:1927–1928
    [Google Scholar]
  39. Woods J.P., Spinola S. M., Strobel S. M., Cannon J. G. 1989; Conserved lipoprotein H.8 of pathogenic Neisseria consists entirely of pentapeptide repeats. Molecular Microbiology 3:43–48
    [Google Scholar]
  40. 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 gonorrhoea. Journal of Infectious Diseases 149:166–173
    [Google Scholar]
  41. Zollinger W. D., Ray J. S., Moran E. E., Seid R. 1985 Identification by monoclonal antibody of an antigen common to the pathogenic Neisseria species. In The Pathogenic Neisseriae: Proceedings of the Fourth International Symposium, pp. 579–584 Edited by Schoolnik G. K. Washington DC: American Society for Microbiology;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-138-11-2321
Loading
/content/journal/micro/10.1099/00221287-138-11-2321
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