1887

Abstract

Lipopolysaccharides (LPS) were extracted from whole cells of seven strains of - 381, ATCC 33277, BH18/10, OMZ314, OMZ409, 6/26 and HW24D-1 - by the phenol/water procedure, and purified by treatment with nuclease and by repeated ultracentrifugation. These LPS were composed of hexoses, hexosamines, fatty acids, phosphorus and phosphorylated 2-keto-3-deoxyoctonate (KDO). The major components of the lipid portion of these LPS were hexadecanoic, 3-hydroxyhexadecanoic, branched 3-hydroxypentadecanoic and branched 3-hydroxyheptadecanoic acids. All the LPS preparations induced marked mitogenic and polyclonal B cell activation responses in spleen cells from both C3H/HeN and C3H/HeJ mice, exhibited no definitive preparatory activity in the local Shwartzman reaction in rabbits, but were active in the chromogenic Limulus amoebocyte lysate test. A monoclonal antibody (mAb) raised against the LPS from strain 6/26 reacted with LPS from all other strains tested. Other mAbs raised against LPS from strains 381 and 6/26 reacted with the LPS from strains 381, ATCC 33277, BH18/10 and 6/26 (these strains were termed LPS serogroup I), as revealed by ELISA and immunodiffusion. The LPS from these strains except for 6/26 showed almost indentical patterns in SDS-PAGE stained with ammoniacal silver. A mAb raised against the LPS from HW24D-1 reacted with the LPS from strains OMZ314, HW24D-1 and OMZ409 (LPS serogroup II). These LPS, except OMZ409, exhibited very similar profiles in SDS-PAGE. These results indicate that there are at least two different antigenic groups present among LPS from strains, as well as a common, species-specific antigen.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-2-319
1990-02-01
2022-01-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/2/mic-136-2-319.html?itemId=/content/journal/micro/10.1099/00221287-136-2-319&mimeType=html&fmt=ahah

References

  1. Beckmann I., Van Eijk H. G., Meisel-Mikolajczyk F., Wallenburg H.C.S. 1989; Detection of 2-keto-3-deoxyoctonate in endotoxins isolated from six reference strains of the Bacteroides fragilis group. International Journal of Biochemistry 21:661–666
    [Google Scholar]
  2. Bligh E. G., Dyer W. J. 1959; A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology 37:911–917
    [Google Scholar]
  3. Cunningham A. J., Szenberg A. 1968; Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology 14:599–600
    [Google Scholar]
  4. Daly C. G., Seymour G. J., Kieser J. B. 1980; Bacterial endotoxin: a role in chronic inflammatory periodontal disease?. Journal of Oral Pathology 9:1–15
    [Google Scholar]
  5. Fujiwara T., Nishihara T., Koga T., Hamada S. 1988; Serological properties and immunobiological activities of lipopolysaccharides from black-pigmented and related oral Bacteroides species. Journal of General Microbiology 134:2867–2876
    [Google Scholar]
  6. Hamada S., Koga T., Nishihara T., Fujiwara T., Okahashi N. 1988; Characterization and immunobiologic activities of lipopolysaccharides from periodontal bacteria. Advances in Dental Research 2:284–291
    [Google Scholar]
  7. Hofstad T. 1974; The distribution of heptose and 2-keto-3-deoxy-octonate in Bacteroidaceae. Journal of General Microbiology 85:314–320
    [Google Scholar]
  8. Johne B., Bryn K. 1986; Chemical composition and biological properties of a lipopolysaccharide from Bacteroides intermedius . Acta Pathologica, Microbiologica et Immunologica Scandinavica B94:265–271
    [Google Scholar]
  9. Karkhanis Y. D., Zeltner J. Y., Jackson J. J., Carlo D. J. 1978; A new and improved microassay to determine 2-keto-3- deoxyoctonate in lipopolysaccharide of Gram-negative bacteria. Analytical Biochemistry 85:595–601
    [Google Scholar]
  10. Kasper D. L., Weintraub A., Lindberg A. A., LÖnngren J. 1983; Capsular polysaccharides and lipopolysaccharides from two Bacteroides fragilis reference strains: chemical and immunochemical characterization. Journal of Bacteriology 153:991–997
    [Google Scholar]
  11. Koga T., Nishihara T., Fujiwara T., Nisizawa T., Okahashi N., Noguchi T., Hamada S. 1985; Biochemical and immunobiological properties of lipopolysaccharide (LPS) from Bacteroides gingivalis and comparison with LPS from Escherichia coli . Infection and Immunity 47:638–647
    [Google Scholar]
  12. Koga T., Odaka C., Moro I., Fujiwara T., Nishihara T., Okahashi N., Hamada S. 1987; Local Shwartzman activity of lipopolysaccharides from several selected strains of suspected periodontopathic bacteria. Journal of Periodontal Research 22:103–107
    [Google Scholar]
  13. Kumada H., Watanabe K., Umemoto T., Haishima Y., Kondo S., Hisatsune K. 1988; Occurrence of O-phosphorylated 2-keto-3- deoxyoctonate in the lipopolysaccharide of Bacteroides gingivalis . FEMS Microbiology Letters 51:77–80
    [Google Scholar]
  14. Mansheim B. J., Onderdonk A. B., Kasper D. L. 1978; Immunochemical and biologic studies of the lipopolysaccharide of Bacteroides melaninogenicus subspecies asaccharolyticus . Journal of Immunology 120:72–78
    [Google Scholar]
  15. Mashimo J., Yoshida M., Ikeuchi K., Hata S., Arata S., Kasai N., Okuda K., Takazoe I. 1985; Fatty acid composition and Shwartzman activity of lipopolysaccharides from oral bacteria. Microbiology and Immunology 29:395–403
    [Google Scholar]
  16. Mayrand D., Holt S. C. 1988; Biology of asaccharolytic black- pigmented Bacteroides species. Microbiological Reviews 52:134–152
    [Google Scholar]
  17. Millar S. J., Chen P. B., Hausmann E. 1987; Monoclonal antibody for identification of Bacteroides gingivalis lipopolysaccharide. Journal of Clinical Microbiology 25:2437–2439
    [Google Scholar]
  18. Morrison D. C. 1983; Bacterial endotoxins and pathogenesis. Reviews of Infectious Diseases 5:S733–S747
    [Google Scholar]
  19. Nair B. C., Mayberry W. R., Dziak R., Chen P. B., Levine M. J., Hausmann E. 1983; Biological effects of a purified lipopolysaccharide from Bacteroides gingivalis . Journal of Periodontal Research 18:40–49
    [Google Scholar]
  20. Okuda K., Ohta K., Kato T., Takazoe I., Slots J. 1986; Antigenic characteristics and serological identification of 10 black- pigmented Bacteroides species. Journal of Clinical Microbiology 24:89–95
    [Google Scholar]
  21. Ryhage R., Stenhagen E. 1960; Mass spectrometric studies. VI. Methyl esters of normal chain oxo-, hydroxy-, methoxy- and epoxyacids. Arkiv för Kemi 15:545–574
    [Google Scholar]
  22. Sawardeker J. S., Sloneker J. H., Jeanes A. 1965; Quantitative determination of monosaccharides as their alditol acetates by gas liquid chromatography. Analytical Chemistry 37:1602–1604
    [Google Scholar]
  23. Schenck K., Michaelsen T. E. 1987; IgG subclass distribution of serum antibodies against lipopolysaccharide from Bacteroides gingivalis in periodontal health and disease. ActaPathologica, Microbiologica et ImmunologicaScandinavicaC 95:41–46
    [Google Scholar]
  24. Schenck K., Helgeland K., Tollefsen T. 1987; Antibodies against lipopolysaccharide from Bacteroides gingivalis before and after periodontal treatment. Scandinavian Journal of Dental Research 95:112–118
    [Google Scholar]
  25. Slots J., Genco R. J. 1984; Black-pigmented Bacteroides species, Capnocytophaga species, and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonization, survival, and tissue destruction. Journal of Dental Research 63:412–421
    [Google Scholar]
  26. Slots J., Listgarten M. A. 1988; Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. Journal of Clinical Periodontology 15:85–93
    [Google Scholar]
  27. Sveen K. 1977; The capacity of lipopolysaccharides from Bacteroides, Fusobacterium, and Veillonella to produce skin inflammation and the local and generalized Shwartzman reaction in rabbits. Journal of Periodontal Research 12:340–350
    [Google Scholar]
  28. Trevelyan W. E., Harrison J. S. 1956; Studies on yeast metabolism. 7. Yeast carbohydrate fractions, separation from nucleic acid, analysis, and behaviour during anaerobic fermentation. Biochemical Journal 63:23–33
    [Google Scholar]
  29. Tsai C. M., Frasch C. E. 1982; A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Analytical Biochemistry 119:115–119
    [Google Scholar]
  30. 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]
  31. Westphal O., Jann K. 1965; Bacterial lipopolysaccharides.Extraction with phenol-water and further applications of the procedure. Methods in Carbohydrate Chemistry 5:83–91
    [Google Scholar]
  32. Westphal O., LÜderitz O., Galanos C., Mayer H., Rietschel E. T. 1985; The story of bacterial endotoxin. In Advances in Immunopharmacology pp. 13–34 Chedid L., Hadden J. W., Spreafico F., Dukor P., Willoughby D. Edited by Oxford: Pergamon Press;
    [Google Scholar]
  33. Wollenweber H. W., Rletschel E. T., Hofstad T., Weintraub A., Lindberg A. A. 1980; Nature, type of linkage, quantity, and absolute configuration of (3-hydroxy) fatty acids in lipopolysaccharides from Bacteroides fragilis NCTC 9343 and related strains. Journal of Bacteriology 144:898–903
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-2-319
Loading
/content/journal/micro/10.1099/00221287-136-2-319
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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