1887

Abstract

Summary

Antibody-binding domains on the major subunits of serotype 2 (Fim2) and 3 fimbriae (Fim3) have been identified using synthetic peptides which were screened for recognition by anti-protein monoclonal antibodies (mAbs). The presence of non-contiguous fimbrial epitopes was demonstrated by both anti-Fim2 and anti-Fim3 mAbs, several of which recognized at least two peptides that were discontinuous in the amino acid sequence of the corresponding subunits. The specificity of one mAb, 51/24, directed against Fim2, was investigated by replacement-set analysis of a 10-residue peptide, and revealed that antibody binding to the peptide was dependent on the sequence NPQ which is non-conserved in Fim3. Furthermore, proline at residue 95 was found to be essential for mAb 51/24 binding. The specific anti-Fim3 mAb, AG3A, was found to recognize the 10-residue carboxy-terminal peptide from both Fim3 and, unexpectedly, from Fim2. This result suggests that mAb AG3A serospecificity at the protein level is determined by a conformational constraint which prevents mAb AG3A binding to the Fim2 C-terminal domain. Several free peptides containing amino acid residues which comprise part of the Fim2 and Fim3 epitopic domains were prepared as immunogens. One of these peptides was immunogenic in the mouse, indicating the location of a T-helper cell epitope within the peptide sequence, and induced a strong anti-peptide antibody response. The other peptides each required immunization as a conjugate with a carrier protein for anti-peptide antibody stimulation. All four anti-peptide antibody preparations only weakly recognized fimbriae-coated ELISA plates. The results of this investigation demonstrate that although short linear peptides can mimic sub-domains of non-contiguous fimbrial epitopes, they are, however, poor candidate antigens for stimulating an anti-fimbrial antibody response.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-1-205
1994-01-01
2021-08-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/1/mic-140-1-205.html?itemId=/content/journal/micro/10.1099/13500872-140-1-205&mimeType=html&fmt=ahah

References

  1. Ashworth L.A.E., Dowsett A.B., Irons L.I., Robinson A. 1985; The location of surface antigens of Bordeiella pertussis by immuno-electron microscopy. Dev Biol Stand 61:143–151
    [Google Scholar]
  2. Cassels F.J., Deal C.D., Reid R.H., Jarboe D.L., Neuss J.L., Carter J.M., Boedeher E.C. 1992; Analysis of Escherichia coli colonisaiion factor antigen I linear B-cell epitopes, as determined by primate responses, following protein sequence verification. Infect Immun 60:2174–2181
    [Google Scholar]
  3. Crumpton M. J. 1986; The importance of conformation and of equilibria in the interaction of globular proteins and their fragments with antibodies. Porter R., Whelan J. Synthetic Peptides as Antigens Chichester: Wiley: Ciba Foundation Symposia; 1993–106
    [Google Scholar]
  4. Frederiksen J.H., Froholm L.O., Kjennerud U. 1987; The specificity of antisera against Bordetella pertussi examined by bacterial agglutination. Acta Pathol Microbiol Immunol Scand B95:363–369
    [Google Scholar]
  5. Getzoff E.D., Geysen H.M., Roddas S.J., Alexander H., Tainer J.A., Lerner R.A. 1987; Mechanisms of antibody binding to a protein. Science 235:1191–1196
    [Google Scholar]
  6. Geysen H.M., Meloen R.H., Barteling S.J. 1984; Use of peptide synthesis to probe viral antigens for epitopes to the resolution of a single amino acid. Proc Natl Acad Set USA 81:3998–4002
    [Google Scholar]
  7. Green N., Alexander H., Olson A., Alexander S., Shinnick T.M., Sutcliffe J.G., Lerner R.A. 1982; Immunogenic structure of the influenza virus haemagglutinin. Cell 28:477–487
    [Google Scholar]
  8. Irons L I., Ashworth L.A.E., Robinson A. 1985; Release and purification of fimbriae from Bordetella pertussis . Dev Biol Stand 61:153–163
    [Google Scholar]
  9. Jemmerson R., Blankenfeld R. 1989; Affinity considerations in the design of synthetic vaccines intended to elicit antibodies. Mol Immunol 26:301–307
    [Google Scholar]
  10. Kent S. B. H. 1988; Chemical synthesis of peptides and proteins. Annu Rev Biochem 57:957–989
    [Google Scholar]
  11. Lamb J.R., Ivanji J., Rees A.D.M., Rothbard J.B., Howland K., Young R., Young D.B. 1987; Mapping of T cell epitopes using recombinant antigens and synthetic peptides. EMBO J 6:1245–1249
    [Google Scholar]
  12. Livey I., Duggleby C.J., Robinson A. 1987; Cloning and nucleotide sequence analysis of the serotype 2 fimbrial subunit gene of Bordetella pertussis . Mol Microbiol 1:203–209
    [Google Scholar]
  13. Mooi F.R., ter Avest A., van der Heide H.G.J. 1990; Structure of the Bordetella pertussis gene coding for the serotype 3 fimbrial subunit. FEMS Microbiol Lett 66:327–332
    [Google Scholar]
  14. Pearce A.M., Irons L I., Robinson A., Seabrook R.N. 1992; Effects of guanidinium hydrochloride on the structure and immunological properties of Bordetella pertussis fimbriae. Biocbem J 283:823–828
    [Google Scholar]
  15. Preston N. W. 1985; Essential immunogens in human pertussis: the role of fimbriae. Dev Biol Stand 61:137–141
    [Google Scholar]
  16. Robinson A., Gorringe A.R., Funnell S.G.P., Fernandez M. 1989; Serospecific protection of mice against intranasal infection with Bordetella pertussis. . Vaccine 7:321–324
    [Google Scholar]
  17. Robinson A., Irons L.I., Seabrook R.N., Pearce A., Matheson M., Funnell S.G.P. 1991; Structure-function studies of Bordetella pertussis fimbriae.. Proceedings of the Sixth International Symposium on Pertussis 126–135 Manclark C. R. Bethesda: FDA: DHHS No. 90-1164;
    [Google Scholar]
  18. Scherf T., Hiller R., Naider F., Levitt M., Anglister J. 1992; Induced peptide conformations in different antibody complexes: molecular modelling of the three dimensional structure of peptide-antibody complexes using NMR-derived distance restraints. Biochemistry 31:6884–6897
    [Google Scholar]
  19. Seabrook R.N., Robinson A., Sharma R.P., Irons L.I., Ashworth L.A.E., Price C.P., Atkinson T. 1990; Recognition of pertussis toxin by antibodies to synthetic peptides. Mol Immunol 27:777–785
    [Google Scholar]
  20. Siligardi G., Drake A.F., Mascagni P., Rowlands D., Brown F., Gibbons W.A. 1991; Correlations between the conformations elucidated by CD spectroscopy and the antigenic properties of four peptides of the foot-and-mouth disease virus. Bur J Biochem 199:545–551
    [Google Scholar]
  21. Stanfield R.L., Fieser T., Lerner R.A., Wilson I.A. 1990; Crystal structures of an antibody to a peptide and its complex with peptide antigen at 2-8 A. Science 248:712–719
    [Google Scholar]
  22. Steven A.C., Bisher M.E., Trus B.L., Thomas D., Zhang J.M., Cowell J.L. 1986; Helical structure of Bordetella pertussis fimbriae. J Bacterial 167:968–974
    [Google Scholar]
  23. Tallet A., Seabrook R.N., Irons L I., Robinson A., van Heyningen S., Atkinson T. 1993; Localisation of a receptor domain on the S3 subunit of pertussis toxin by peptide mapping. Bur J Biocbem 211:743–748
    [Google Scholar]
  24. Willems R.J., van der Heide H.G. J., Mooi F.R. 1992; Characterisation of a Bordetella pertussis fimbrial gene cluster which is located directly downstream of the filamentous haemagglutinin gene. Mol Microbiol 6:2661–2671
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-1-205
Loading
/content/journal/micro/10.1099/13500872-140-1-205
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