1887

Abstract

The association of staphylococci with the mucus gel that overlays the mucosa of the respiratory tract may lead to clearance of cocci or, in certain conditions such as cystic fibrosis (CF), to colonization. In the present study, a quantitative radioassay was used to study the effect of Ca, which is elevated in CF sputa, on the adhesion of H-labelled to submaxillary gland mucin immobilized in MaxiSorp 96-well, break-apart modules. Ca significantly enhanced the adhesion of (five strains) and (four strains). The reaction was specific because adhesion was not enhanced in the presence of Mg, Ca + EGTA (a Ca chelator) or protamine and was not attributable to hydrophobicity of the test strains. Staphylococcal adhesion was significantly (⩽0·005) blocked in the presence of highly sialated and sulphated reagents, which suggests that Ca binds to the sialic acid and sulphate residues of immobilized mucin. The Ca -binding sites on the surface of were trypsin-sensitive; in addition, I-labelled solubilized surface proteins reacted with immobilized mucin in a direct binding assay, and the reaction was significantly enhanced by Ca. Autoradiography demonstrated that Ca bound directly to two polypeptides ( 170000 and 150000) of solubilized staphylococcal surface proteins separated by SDS-PAGE, and that I-labelled mucin bound directly to three staphylococcal polypeptides ( 40000, 35000, and 29000). These results suggest that adhesion to mucin is mediated by at least two mechanisms: via Ca-binding surface proteins in the presence of Ca and via mucin-binding surface proteins unrelated to Ca.

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1993-03-01
2024-04-23
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References

  1. Bauernfeind A., Bertele R. M., Harms K., Horl G., Jungwirth R., Petermuller C., Przyklenk B., Weisslein-Pfister C. 1987; Qualitative and quantitative microbiological analysis of sputa of 102 patients with cystic fibrosis. Infection 15:270–277
    [Google Scholar]
  2. Biesbrock A. R., Reddy M. S., Levine M. J. 1991; Interaction of a salivary mucin-secretory immunoglobulin A complex with mucosal pathogens. Infection and Immunity 10:3492–3497
    [Google Scholar]
  3. Burns M. W., May J. R. 1968; Bacterial precipitins in serum of patients with cystic fibrosis. Lancet 1:270–272
    [Google Scholar]
  4. Cohen P. S., Arruda J. C., Williams T. J., Laux D. C. 1985; Adhesion of a human fecal Escherichia coli strain to mouse colonic mucus. Infection and Immunity 48:139–145
    [Google Scholar]
  5. Daugharty D., Warfield D. T., Davis M. L. 1972; Solid-phase radioimmunoassay of total and influenza-specific immunoglobulin. General Applied Microbiology 23:360–367
    [Google Scholar]
  6. Dzandu J. K., Deh M. E., Barratt D. L., Wise G. E. 1984; Detection of erythrocyte membrane proteins, sialoglycoproteins, and lipids in the same polyacrylamide gel using a double-staining technique. Proceedings of the National Acadademy of Sciences of the United States of America 81:1733–1737
    [Google Scholar]
  7. Florey H. 1955; Mucin and the protection of the body. Proceedings of the Royal Society of London 143:147–159
    [Google Scholar]
  8. Forsgren A., Sjoquist J. 1966; ‘Protein A’ from S. aureus. I. Pseudo-immune reaction with human λ-globulin. Journal of Immunology 97:822–827
    [Google Scholar]
  9. Forstner J. F., Forstner G. G. 1975; Calcium binding to intestinal goblet cell mucin. Biochimica et Biophysica Acta 386:283–292
    [Google Scholar]
  10. Freter R. 1980; Prospects for preventing the association of harmful bacteria with host mucosal surfaces. In Bacterial Adherence pp. 441–458 Beachey E. H. Edited by London: Chapman Hall;
    [Google Scholar]
  11. Goerging R. V., Bauernfeind A., Lenz W., Przyklenk B. 1990; Staphylococcus aureus in patients with cystic fibrosis: an epidemiological analysis using a combination of traditional and molecular methods. Infection 18:57–60
    [Google Scholar]
  12. Herrmann M., Suchard S. J., Boxer L. A., Waldvogel F. A., Lew P. D. 1991; Thrombospondin binds to Staphylococcus aureus and promotes staphylococcal adherence to surfaces. Infection and Immunity 59:279–288
    [Google Scholar]
  13. Kilbourn J. P. 1978; Bacterial content and ionic composition of sputum in cystic fibrosis. Lancet i:334
    [Google Scholar]
  14. Kilbourn J. P. 1984; Composition of sputum from patients with cystic fibrosis. Current Microbiology 11:19–22
    [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  16. Liang O. D., Ascencio F., Fransson L. A., Wadstrom T. 1992; Binding of heparan sulphate to Staphylococcus aureus. Infection and Immunity 60:899–906
    [Google Scholar]
  17. Mcnabb P. C. 1981; Host defense mechanisms at mucosal surfaces. Annual Review of Microbiology 35:477–496
    [Google Scholar]
  18. Marks M. I. 1990; Staphylococcal aspects of cystic fibrosis. Infection 18:53–56
    [Google Scholar]
  19. Maruyama K., Mikawa T., Ebashi S. 1984; Detection of calcium binding proteins by 45Ca autoradiography on nitrocellulose membrane after sodium dodecyl sulphate gel electrophoresis. Journal of Biochemistry 95:511–519
    [Google Scholar]
  20. Nisizawa K., Pigman W. 1959; The composition and properties of the mucin clot from cattle submaxillary glands. Archives of Oral Biology 1:161–170
    [Google Scholar]
  21. Patel A. H., Nowlan P., Weavers E. D., Foster T. 1987; Virulence of protein A-deficient and alpha-toxin-deficient mutants of Staphylococcus aureus isolated by allele replacement. Infection and Immunity 55:3103–3110
    [Google Scholar]
  22. Ramphal R. 1990; The role of bacterial adhesion in cystic fibrosis including the staphylococcal aspect. Infection 18:61–64
    [Google Scholar]
  23. Roberts G. P. 1978; Chemical aspects of respiratory mucus. British Medical Bulletin 34:39–41
    [Google Scholar]
  24. Rosenberg M., Gutnick D., Rosenberg E. 1980; Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiology Letters 9:29–33
    [Google Scholar]
  25. Sanford B. A., Ramsay M. A. 1987; Bacterial adherence to the upper respiratory tractof ferrets infected with influenza A virus. Proceedings of the Society for Experimental Biology and Medicine 181:104–111
    [Google Scholar]
  26. Sanford B. A., Thomas V. L., Ramsay M. A., Jones T. O. 1986; Characterization of clinical strains of Staphylococcus aureus associated with pneumonia. Journal of Clinical Microbiology 24:131–136
    [Google Scholar]
  27. Sanford B. A., Thomas V. L., Ramsay M. A. 1989; Binding of Staphylococci to mucus in vivo and in vitro. Infection and Immunity 57:3735–3742
    [Google Scholar]
  28. Schadow K. H., Simpson W. A., Christensen G. D. 1988; Characteristics of adherence to plastic tissue culture plates of coagulase-negative staphylococci exposed to subinhibitory concentrations of antimicrobial agents. Journal of Infectious Diseases 157:71–77
    [Google Scholar]
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