Immunochemical Characterization of a Polysaccharide Antigen of with an IgM Monoclonal Antibody Free

Abstract

An IgM mouse monoclonal antibody (McAb) Bf4 was produced to a surface polysaccharide of NCTC 9343. Immunoblotting showed that McAb Bf4 reacted strongly with a high molecular mass structure which was sensitive to oxidation with periodate but resisted protease treatment. An inhibition enzyme-linked immunosorbent assay (ELISA) indicated that McAb Bf4 did not cross react with the sixteen species and strains tested. Cells of NCTC 9343 recovered from the various interfaces of a Percoll discontinuous density gradient were tested in the inhibition ELISA. Bacteria from the 0–20 %, 20–40 % and 40–60 % interfaces inhibited the ELISA; however, cells from the 60–80 % interface did not. Electron microscopy with immunogold labelling showed that McAb Bf4 did not react with the extracellular fibrous network on bacteria recovered from the 0–20 % interface, or the extracellular electron dense layer on cells from the 60–80 % interface; however, it was associated with a surface structure on cells from the 20–40 % interface. Growth did not enrich for bacteria with this structure.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-133-1-171
1987-01-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/133/1/mic-133-1-171.html?itemId=/content/journal/micro/10.1099/00221287-133-1-171&mimeType=html&fmt=ahah

References

  1. Babb J. I., Cummins C. S. 1978; Encapsulation of Bacteroides species. Infection and Immunity 19:1088–1093
    [Google Scholar]
  2. Connolly J. C., Mcclean C., Tabaqchali S. 1984; The effect of capsular polysaccharide of Bacteroides fragilis on polymorph function and serum killing. Journal of Medical Microbiology 17:259–271
    [Google Scholar]
  3. Cousland G., Poxton I. R. 1983; Analysis of lipopolysaccharides of Bacteroides fragilis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electroblot transfer. FEMS Microbiology Letters 20:461–465
    [Google Scholar]
  4. Cousland G., Poxton I. R. 1984; Crossed immunoelectrophoresisand enzyme-linked immunosorbent assay of the cell-surface antigens of Bacteroides fragilis. Journal of General Microbiology 130:645–655
    [Google Scholar]
  5. Duerden B. I. 1980; The identification of Gram negative anaerobic bacilli isolated from clinical infections. Journal of Hygiene 84:69
    [Google Scholar]
  6. Galanos C., Luderitz E., Westphal O. 1969; A new method for the extraction of R lipopolysaccharides. European Journal of Biochemistry 9:245–249
    [Google Scholar]
  7. Gorbach L. S., Bartlett J. G. 1974; Anaerobic infections. New England Journal of Medicine 290:1177–1184
    [Google Scholar]
  8. Hofstad T. 1984; Pathogenicity of anaerobic Gramnegative rods: possible mechanisms. Reviews of Infectious Diseases 6:189–199
    [Google Scholar]
  9. Kasper D. L. 1976; The polysaccharide capsule of Bacteroides fragilis subspecies fragilis: immunochemical and morphological definition. Journal of Infectious Diseases 133:79–87
    [Google Scholar]
  10. Kasper D. L., Onderdonk A. B., Reinap B. G., Lindberg A. A. 1980; Variations of Bacteroides fragilis with in vitro passage: presence of an outer membrane-associated glycan and loss of capsular antigen. Journal of Infectious Diseases 142:750–756
    [Google Scholar]
  11. Kasper D. L., Weintraub A. B., Lindberg A. A., Lonngren J. 1983; Capsular polysaccharides and lipopolysaccharides from two Bacteroides fragilis reference strains: chemical and immunochemical characterization. Journal of Bacteriology 153:991–997
    [Google Scholar]
  12. Lambe D. W., Mayberry-Carson K. J. S., Ferguson K. P., Costerson J. W. 1984; Morphological stabilization of the glycocalyces of 23 strains of five Bacteroides species using specific antisera. Canadian Journal of Microbiology 30:809–819
    [Google Scholar]
  13. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  14. Lemke H., Hammerling G. J., Hohmann C., Rajewsky K. 1978; Hybrid cell lines secreting monoclonal antibody specific for major histocompatibility antigens of the mouse. Nature; London: 271249–251
    [Google Scholar]
  15. Linko-Kettunen L., Arstila P., Jalkanen M., Jousimies-Somer H., Lassila O., Lehtonen O. P., Weintraub A., Viljanen M. K. 1984; Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide. Journal of Clinical Microbiology 20:519–524
    [Google Scholar]
  16. Morris D. L. 1948; Quantitative determination of carbohydrates with Dreywood’s anthrone reagent. Science 107:254–255
    [Google Scholar]
  17. Onderdonk A. B., Kasper D. L., Cisneros R. L., Bartlett J. G. 1977; The capsular polysaccharide of Bacteroides fragilis as a virulence factor: comparison of the pathogenic potential of encapsulated and unencapsulated strains. Journal of Infectious Diseases 136:82–89
    [Google Scholar]
  18. Patrick S., Reid J. H. 1983; Separation of capsulate and non-capsulate Bacteroides fragilis on a discontinuous density gradient. Journal of Medical Microbiology 16:239–241
    [Google Scholar]
  19. Patrick S., Reid J. H., Larkin M. J. 1984; The growth and survival of capsulate and non-capsulate Bacteroides fragilis in vivo and in vitro. Journal of Medical Microbiology 17:237–246
    [Google Scholar]
  20. Patrick S., Reid J. H., Coffey A. 1986; Capsulation of in vitro and in vivo grown Bacteroides species. Journal of General Microbiology 132:1099–1109
    [Google Scholar]
  21. Peterson A. A., Mcgroarty E. J. 1985; High-molecular-weight components in lipopolysaccharides of Salmonella typhimurium, Salmonella minnesota and Escherichia coli. Journal of Bacteriology 162:738–745
    [Google Scholar]
  22. Poxton I. R., Bell G. T., Barclay G. R. 1985; The association on SDS-polyacrylamide gels of lipopolysaccharide and outer membrane proteins of Pseudomonas aeruginosa as revealed by monoclonal antibodies and Western blotting. FEMS Microbiology Letters 27:247–251
    [Google Scholar]
  23. Reid J. H., Patrick S., Dermott E., Trudgett A., Tabaqchali S. 1985; Investigation of antigenic expression of Bacteroides fragilis by immunogold labelling and immunoblotting with a monoclonal antibody. FEMS Microbiology Letters 30:289–293
    [Google Scholar]
  24. van Tassell R. L., Wilkins T. D. 1978; Isolation of auxotrophs of Bacteroides fragilis. Canadian Journal of Microbiology 24:1619–1621
    [Google Scholar]
  25. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
    [Google Scholar]
  26. Weintraub A., Larsson B. E., Lindberg A. A. 1985; Chemical and immunochemical analyses of Bacteroides fragilis lipopolysaccharides. Infection and Immunity 49:197–201
    [Google Scholar]
  27. Westphal O., Luderitz O. 1954; Chemische Erforschung von lipopolysacchariden Gram- negativer Bakterien. Angewandte Chemie 66:407–417
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-133-1-171
Loading
/content/journal/micro/10.1099/00221287-133-1-171
Loading

Data & Media loading...

Most cited Most Cited RSS feed