Ultrastructure, Serogrouping and Localization of Surface Antigens of Free

Abstract

The surface ultrastructure of 21 strains of was investigated by electron microscopy. Rat monoclonal antibodies (mAbs) were used to define serogroups and to detect the location of surface antigens. All 21 isolates had capsules as demonstrated by the use of wet and dry Indian ink stains. Negative staining of whole cells with 1% (w/v) methylamine tungstate showed that all 21 isolates carried clumped peritrichous fibrils with strain dependent morphology, density and length (≤0·75 μm). Fibrils on 11 of 13 fresh clinical isolates were more conspicuously clumped and easily visible, whereas those on 6 of 8 laboratory strains were indistinct and were at the limits of the resolution of the negative staining technique. Staining with ruthenium red (), followed by thin sectioning, revealed a dense, amorphous staining layer (), up to 24·8 ±3·0 nm thick, adjacent to the outer membrane on all of 15 strains examined. All isolates had a less dense staining matrix () extending away from the The structure of the varied between strains. Four rat mAbs (376.1, 381, 391.1 and 403.2) were used to serogroup the 21 strains of Immunonegative staining revealed that the mAbs were not directed against fibrils. Antigens recognized by mAb 376.1 and mAb 391.1 were located on the surfaces of cells, beneath fibrils, and on extracellular vesicles. mAb 381 recognized an antigen which was most accessible on lysed cells, and non-specific binding of mAb 403.2 to grids prevented its localization on the cell surface.

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1989-04-01
2024-03-29
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References

  1. Bayer M.E., Thurow H. 1977; Polysaccharide capsule of Escherichia coli Microscope study of its size, structure and sites of synthesis. Journal of Bacteriology 130:911–936
    [Google Scholar]
  2. Costerton J.W., Irvin R.T., Cheng K.-J. 1981; The bacterial glycocalyx in nature and disease. Annual Review of Microbiology 35:299–324
    [Google Scholar]
  3. GmÜr R., Guggenheim B. 1983; Antigenic heterogeneity of Bacteroides intermedius as recognized by monoclonal antibodies. Infection and Immunity 42:459–470
    [Google Scholar]
  4. GmÜr R., Wyss C. 1985; Monoclonal antibodies to characterize the antigenic heterogeneity of Bacteroides intermedius . In Monoclonal Antibodies against Bacteria 1 pp. 91–119 Macario A. J. L., Conway de Macario E. Edited by London: Academic Press;
    [Google Scholar]
  5. Grenier D., Mayrand D. 1987; Functional characterization of extracellular vesicles produced by Bacteroides gingivalis . Infection and Immunity 55:111–117
    [Google Scholar]
  6. GrÜneberg R.N., Leigh D.A., Brumfitt W. 1968; Escherichia coli serotypes in urinary tract infection; studies in domiciliary, antenatal and hospital practice. In Urinary Tract Infection pp. 68–79 O’Grady F., Brumfitt W. Edited by Oxford: Oxford University Press;
    [Google Scholar]
  7. Handley P.S., Tipler L.S. 1986; An electron microscope survey of the surface structures and hydrophobicity of oral and non-oral species of the bacterial genus Bacteroides . Archives of Oral Biology 31:325–335
    [Google Scholar]
  8. Handley P.S., Carter P.L., Fielding J. 1984; Streptococcus salivarius strains carry either fibrils or fimbriae on the cell surface. Journal of Bacteriology 157:64–72
    [Google Scholar]
  9. Handley P.S., Carter P.L., Wyatt J.E., Hesketh L.M. 1985; Surface structures (peritrichous fibrils and tufts of fibrils) found on Streptococcus sanguis strains may be related to their ability to coaggregate with other oral genera. Infection and Immunity 47:217–227
    [Google Scholar]
  10. Handley P.S., Hargreaves J., Harty D.W.S. 1988; Ruthenium red staining reveals surface fibrils and a layer external to the cell wall in Streptococcus salivarius HB and adhesion deficient mutants. Journal of General Microbiology 134:3165–3172
    [Google Scholar]
  11. Hesketh L.M., Wyatt J.E., Handley P.S. 1987; Effect of protease on cell surface structure, hydrophobicity and adhesion of tufted strains of Streptococcus sanguis biotypes I and II. Microbios 50:131–145
    [Google Scholar]
  12. Hoekstra D., Van Der Laan J.W., De Leij L., Without B. 1976; Release of outer membrane fragments from normally growing Escherichia coli . Biochimica et biophysica acta 455:889–899
    [Google Scholar]
  13. Johnson J.L., Holdeman L.V. 1983; Bacteroides intermedius comb. nov. and descriptions of Bacteroides corporis sp. nov. and Bacteroides levii sp. nov. International Journal of Systematic Bacteriology 33:15–25
    [Google Scholar]
  14. Kasper D.L. 1976; Chemical and biological characterization of the lipopolysaccharide of Bacteroides fragilis subspecies fragilis . Journal of Infectious Diseases 134:59–66
    [Google Scholar]
  15. Kasper D.L., Hayes M.E., Reinap B.G., Craft F.O., Onderdonk A.B., Pock B.F. 1977; Isolation and identification of encapsulated strains of Bacteroides fragilis . Journal of Infectious Diseases 136:75–81
    [Google Scholar]
  16. Kasper D.L., Onderdonk A.B., Reinap B.G., Lindberg A.A. 1980; Variation 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]
  17. Knox K.W., Vesk M., Work E. 1966; Relation between excreted lipopolysaccharide complexes and surface structures of a lysine-limited culture of Escherichia coli . Journal of Bacteriology 92:1206–1217
    [Google Scholar]
  18. Lai C.-H., Listgarten M.A., Hammond B.F. 1981; Comparative ultrastructure of leukotoxic and non-leukotoxic strains of Actinobacillus actinomycetemcomitans . Journal of Periodontal Research 16:379–389
    [Google Scholar]
  19. Lambe D.W., Mayberry-Carson K.J., Ferguson K.P., Costerton J.W. 1984; Morphological stabilization of the glycocalyx of 23 strains of 5 Bacteroides species using specific antisera. Canadian Journal of Microbiology 30:809–819
    [Google Scholar]
  20. Lantz M.S., Switalski L.M., Kornman K.S., HÖÖK M. 1985; Bacteroides intermedius binds fibrinogen. Journal of Bacteriology 163:623–628
    [Google Scholar]
  21. Listgarten M.A., Lai C.-H. 1979; Comparative ultrastructure of Bacteroides melaninogenicus subspecies. Journal of Periodontal Research 14:332–340
    [Google Scholar]
  22. Loesche W.J., Hockett R.N., Syed S.A. 1972; The predominant cultivable flora of tooth surface plaque removed from institutionalized subjects. Archives of Oral Biology 17:1311–1325
    [Google Scholar]
  23. Luft J.H. 1971; Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anatomical Record 171:347–368
    [Google Scholar]
  24. Mckee A.S., Mcdermid A.S., Baskerville A., Dowsett A.B., Ellwood D.C., Marsh P.D. 1986; Effect of hemin on the physiology and virulence of Bacteroides gingivalis W50. Infection and Immunity 52:349–355
    [Google Scholar]
  25. Mansheim B.J., Kasper D.L. 1977; Purification and immunochemical characterization of the outer membrane complex of Bacteroides melaninogenicus subspecies asaccharolyticus . Journal of Infectious Diseases 135:787–799
    [Google Scholar]
  26. Mayrand D., Holt S.C. 1988; Biology of asaccharolytic black-pigmented Bacteroides species. Microbiological Reviews 52:134–152
    [Google Scholar]
  27. Moore W.E.C., Holdeman L.V., Cato E.P., Smibert R.M., Burmeister J.A., Ranney R.R. 1983; Bacteriology of moderate (chronic) periodontitis in mature adult humans. Infection and Immunity 42:510–515
    [Google Scholar]
  28. Moore W.E.C., Holdeman L.V., Cato E.P., Smibert R.M., Burmeister J.A., Palcanis K.G., Ranney R.R. 1985; Comparative bacteriology of juvenile periodontitis. Infection and Immunity 48:507–519
    [Google Scholar]
  29. Nakazawa F., Zambon J.J., Reynolds H.S., Genco R.J. 1988; Serological studies of oral Bacteroides intermedius . Infection and Immunity 56:1647–1651
    [Google Scholar]
  30. Nowotny A., Behling U.H., Hammond B., Lai C.-H., Listgarten M., Pham P.H., Sanavi F. 1982; Release of toxic microvesicles by Actinobacillus actinomycetemcomitans . Infection and Immunity 37:151–154
    [Google Scholar]
  31. Okuda K., Slots J., Genco R.J. 1981; Bacteroides gingivalis, Bacteroides asaccharolyticus and Bacteroides melaninogenicus subspecies: cell surface morphology and adherence to erythrocytes and human buccal epithelial cells. Current Microbiology 6:7–12
    [Google Scholar]
  32. Ottow J.C.G. 1975; Ecology, physiology and genetics of fimbriae and pili. Annual Review of Microbiology 29:79–108
    [Google Scholar]
  33. 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]
  34. Reid J.H., Patrick S. 1984; Phagocytic and serum killing of capsulate and non-capsulate Bacteroides fragilis . Journal of Medical Microbiology 17:247–257
    [Google Scholar]
  35. Reynolds E.S. 1963; The use of lead citrate at high pH as an electron opaque stain in electron microscopy. Journal of Cellular Biology 17:208–212
    [Google Scholar]
  36. Sarff L.D., Mccracken G.H., Schiffer M.S., Glode M.P., Robbins J.B., Ørskov I., Ørskov F. 1975; Epidemiology of Escherichia coli K1 in healthy and diseased newborns. Lancet i:1099–1104
    [Google Scholar]
  37. Slots J. 1979; Subgingival microflora and periodontal disease. Journal of Clinical Periodontology 6:351–382
    [Google Scholar]
  38. Slots J. 1986; Bacterial specificity in adult periodontitis - a summary of recent work. Journal of Clinical Periodontology 13:912–917
    [Google Scholar]
  39. 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]
  40. Van Steenbergen T.J.M., De Graaff J. 1986; Proteolytic activity of black-pigmented Bacteroides strains. FEMS Microbiology Letters 33:219–222
    [Google Scholar]
  41. Van Steenbergen T.J.M., Vlaanderen C.A., De Graaff J. 1982; Deoxyribonucleic acid homologies among strains of Bacteroides melaninogenicus and related species. Journal of Applied Bacteriology 53:269–276
    [Google Scholar]
  42. Thompson S.S., Naidu Y.M., Pestka J.J. 1985; Extracellular localisation of an extracellular protease in Pseudomonas fragi by using the peroxidase-antiperoxidase reaction. Applied and Environmental Microbiology 50:1038–1042
    [Google Scholar]
  43. Weerkamp A.H., Jacobs T. 1982; Cell wall protein antigens of Streptococcus salivarius: purification, properties, and function in adherence. Infection and Immunity 38:233–242
    [Google Scholar]
  44. Woo D.D.L., Holt S.C., Leadbetter E.R. 1979; Ultrastructure of Bacteroides species: Bacteroides asaccharolyticus, Bacteroides fragilis, Bacteroides melaninogenicus subspecies melaninogenicus, and B. melaninogenicus subspecies intermedius . Journal of Infectious Diseases 139:534–546
    [Google Scholar]
  45. Yamamoto A., Takahashi M., Takamori K., Sasaki T. 1982; Ultrastructure of the outer membrane surface of black-pigmented Bacteroides isolated from the human oral cavity. Bulletin of the Tokyo Dental College 23:47–60
    [Google Scholar]
  46. Zambon J.J., Reynolds H.S., Slots J. 1981; Black-pigmented Bacteroides spp. in the human oral cavity. Infection and Immunity 32:198–203
    [Google Scholar]
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