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. 1989 Dec;57(12):3828–3833. doi: 10.1128/iai.57.12.3828-3833.1989

Inhibition of pertussis toxin binding to model receptors by antipeptide antibodies directed at an antigenic domain of the S2 subunit.

M A Schmidt 1, W Schmidt 1
PMCID: PMC259912  PMID: 2478479

Abstract

Receptor recognition of pertussis toxin is mediated by the B-oligomer consisting of subunits S2, S3, S4 (two), and S5. To understand the structure-function relationships of the receptor-binding oligomer and to elucidate the immunological structure of pertussis toxin, we assayed antisera generated against each of 10 synthetic peptides corresponding to segments of the pertussis toxin S2 subunit for their ability to recognize the native toxin. Only antisera raised against peptides R1-7, R35-50, and R91-106 recognized pertussis toxin in an enzyme-linked immunosorbent assay and Western blotting (immunoblotting). These segments thus correspond to linear antigenic epitopes. The highly homologous S3 subunit was only weakly recognized by antibodies to R91-106 in Western blotting. The ability of affinity-purified antipeptide antibodies to interfere with the binding of pertussis toxin was investigated by pertussis toxin-mediated hemagglutination of goose erythrocytes, the binding of pertussis toxin to fetuin, and the toxin-induced clustered growth pattern of CHO cells as model receptor systems. Antibodies directed against synthetic peptides R1-7, R35-50, and R91-106 inhibited the binding of pertussis toxin in the two model receptor systems that solely depend on the interactions of the S2 subunit. The toxin-mediated clustered growth pattern of CHO cells could not be inhibited. The results point to a second binding site with distinct specificity involving the S3 subunit of pertussis toxin. The regions identified in this work contribute to the definition of receptor-binding sites of pertussis toxin and should thus improve the development of an acellular-component pertussis vaccine.

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Selected References

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  1. Arciniega J. L., Burns D. L., Garcia-Ortigoza E., Manclark C. R. Immune response to the B oligomer of pertussis toxin. Infect Immun. 1987 May;55(5):1132–1136. doi: 10.1128/iai.55.5.1132-1136.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Black W. J., Munoz J. J., Peacock M. G., Schad P. A., Cowell J. L., Burchall J. J., Lim M., Kent A., Steinman L., Falkow S. ADP-ribosyltransferase activity of pertussis toxin and immunomodulation by Bordetella pertussis. Science. 1988 Apr 29;240(4852):656–659. doi: 10.1126/science.2896387. [DOI] [PubMed] [Google Scholar]
  3. Brennan M. J., David J. L., Kenimer J. G., Manclark C. R. Lectin-like binding of pertussis toxin to a 165-kilodalton Chinese hamster ovary cell glycoprotein. J Biol Chem. 1988 Apr 5;263(10):4895–4899. [PubMed] [Google Scholar]
  4. Capiau C., Petre J., Van Damme J., Puype M., Vandekerckhove J. Protein-chemical analysis of pertussis toxin reveals homology between the subunits S2 and S3, between S1 and the A chains of enterotoxins of Vibrio cholerae and Escherichia coli and identifies S2 as the haptoglobin-binding subunit. FEBS Lett. 1986 Aug 18;204(2):336–340. doi: 10.1016/0014-5793(86)80839-0. [DOI] [PubMed] [Google Scholar]
  5. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  6. Friguet B., Djavadi-Ohaniance L., Pages J., Bussard A., Goldberg M. A convenient enzyme-linked immunosorbent assay for testing whether monoclonal antibodies recognize the same antigenic site. Application to hybridomas specific for the beta 2-subunit of Escherichia coli tryptophan synthase. J Immunol Methods. 1983 Jun 10;60(3):351–358. doi: 10.1016/0022-1759(83)90292-2. [DOI] [PubMed] [Google Scholar]
  7. Gillenius P., Jätmaa E., Askelöf P., Granström M., Tiru M. The standardization of an assay for pertussis toxin and antitoxin in microplate culture of Chinese hamster ovary cells. J Biol Stand. 1985 Jan;13(1):61–66. doi: 10.1016/s0092-1157(85)80034-2. [DOI] [PubMed] [Google Scholar]
  8. Griffiths E., Kreeftenberg J. G. Developments in pertussis vaccines: memorandum from a WHO meeting. Bull World Health Organ. 1985;63(2):241–248. [PMC free article] [PubMed] [Google Scholar]
  9. Hewlett E. L., Cowell J. L. Evaluation of the mouse model for study of encephalopathy in pertussis vaccine recipients. Infect Immun. 1989 Mar;57(3):661–663. doi: 10.1128/iai.57.3.661-663.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hewlett E. L., Sauer K. T., Myers G. A., Cowell J. L., Guerrant R. L. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. doi: 10.1128/iai.40.3.1198-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Locht C., Barstad P. A., Coligan J. E., Mayer L., Munoz J. J., Smith S. G., Keith J. M. Molecular cloning of pertussis toxin genes. Nucleic Acids Res. 1986 Apr 25;14(8):3251–3261. doi: 10.1093/nar/14.8.3251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Locht C., Cieplak W., Marchitto K. S., Sato H., Keith J. M. Activities of complete and truncated forms of pertussis toxin subunits S1 and S2 synthesized by Escherichia coli. Infect Immun. 1987 Nov;55(11):2546–2553. doi: 10.1128/iai.55.11.2546-2553.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Locht C., Keith J. M. Pertussis toxin gene: nucleotide sequence and genetic organization. Science. 1986 Jun 6;232(4755):1258–1264. doi: 10.1126/science.3704651. [DOI] [PubMed] [Google Scholar]
  15. Monack D., Munoz J. J., Peacock M. G., Black W. J., Falkow S. Expression of pertussis toxin correlates with pathogenesis in Bordetella species. J Infect Dis. 1989 Feb;159(2):205–210. doi: 10.1093/infdis/159.2.205. [DOI] [PubMed] [Google Scholar]
  16. Montecucco C., Tomasi M., Schiavo G., Rappuoli R. Hydrophobic photolabelling of pertussis toxin subunits interacting with lipids. FEBS Lett. 1986 Jan 6;194(2):301–304. doi: 10.1016/0014-5793(86)80105-3. [DOI] [PubMed] [Google Scholar]
  17. Muller A. S., Leeuwenburg J., Pratt D. S. Pertussis: epidemiology and control. Bull World Health Organ. 1986;64(2):321–331. [PMC free article] [PubMed] [Google Scholar]
  18. Munoz J. J., Peacock M. G., Hadlow W. J. Anaphylaxis or so-called encephalopathy in mice sensitized to an antigen with the aid of pertussigen (pertussis toxin). Infect Immun. 1987 Apr;55(4):1004–1008. doi: 10.1128/iai.55.4.1004-1008.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nicosia A., Bartoloni A., Perugini M., Rappuoli R. Expression and immunological properties of the five subunits of pertussis toxin. Infect Immun. 1987 Apr;55(4):963–967. doi: 10.1128/iai.55.4.963-967.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nicosia A., Perugini M., Franzini C., Casagli M. C., Borri M. G., Antoni G., Almoni M., Neri P., Ratti G., Rappuoli R. Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4631–4635. doi: 10.1073/pnas.83.13.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pittman M., Furman B. L., Wardlaw A. C. Bordetella pertussis respiratory tract infection in the mouse: pathophysiological responses. J Infect Dis. 1980 Jul;142(1):56–66. doi: 10.1093/infdis/142.1.56. [DOI] [PubMed] [Google Scholar]
  22. Pittman M. Pertussis toxin: the cause of the harmful effects and prolonged immunity of whooping cough. A hypothesis. Rev Infect Dis. 1979 May-Jun;1(3):401–412. doi: 10.1093/clinids/1.3.401. [DOI] [PubMed] [Google Scholar]
  23. Pittman M. The concept of pertussis as a toxin-mediated disease. Pediatr Infect Dis. 1984 Sep-Oct;3(5):467–486. doi: 10.1097/00006454-198409000-00019. [DOI] [PubMed] [Google Scholar]
  24. Pizza M., Bartoloni A., Prugnola A., Silvestri S., Rappuoli R. Subunit S1 of pertussis toxin: mapping of the regions essential for ADP-ribosyltransferase activity. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7521–7525. doi: 10.1073/pnas.85.20.7521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ralston S., Sarin V., Thanh H. L., Rivier J., Fox J. L., Lindstrom J. Synthetic peptides used to locate the alpha-bungarotoxin binding site and immunogenic regions on alpha subunits of the nicotinic acetylcholine receptor. Biochemistry. 1987 Jun 16;26(12):3261–3266. doi: 10.1021/bi00386a004. [DOI] [PubMed] [Google Scholar]
  26. Robinson A., Irons L. I., Ashworth L. A. Pertussis vaccine: present status and future prospects. Vaccine. 1985 Mar;3(1):11–22. doi: 10.1016/0264-410x(85)90004-0. [DOI] [PubMed] [Google Scholar]
  27. Sato H., Sato Y. Bordetella pertussis infection in mice: correlation of specific antibodies against two antigens, pertussis toxin, and filamentous hemagglutinin with mouse protectivity in an intracerebral or aerosol challenge system. Infect Immun. 1984 Nov;46(2):415–421. doi: 10.1128/iai.46.2.415-421.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schmidt W., Schmidt M. A. Mapping of linear B-cell epitopes of the S2 subunit of pertussis toxin. Infect Immun. 1989 Feb;57(2):438–445. doi: 10.1128/iai.57.2.438-445.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shinnick T. M., Sutcliffe J. G., Green N., Lerner R. A. Synthetic peptide immunogens as vaccines. Annu Rev Microbiol. 1983;37:425–446. doi: 10.1146/annurev.mi.37.100183.002233. [DOI] [PubMed] [Google Scholar]
  30. Tamura M., Nogimori K., Murai S., Yajima M., Ito K., Katada T., Ui M., Ishii S. Subunit structure of islet-activating protein, pertussis toxin, in conformity with the A-B model. Biochemistry. 1982 Oct 26;21(22):5516–5522. doi: 10.1021/bi00265a021. [DOI] [PubMed] [Google Scholar]
  31. Weiss A. A., Hewlett E. L., Myers G. A., Falkow S. Pertussis toxin and extracytoplasmic adenylate cyclase as virulence factors of Bordetella pertussis. J Infect Dis. 1984 Aug;150(2):219–222. doi: 10.1093/infdis/150.2.219. [DOI] [PubMed] [Google Scholar]
  32. Weiss A. A., Hewlett E. L. Virulence factors of Bordetella pertussis. Annu Rev Microbiol. 1986;40:661–686. doi: 10.1146/annurev.mi.40.100186.003305. [DOI] [PubMed] [Google Scholar]

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